Document Information

PMID 14511332  (  )
Title Expression and localization of cyclooxygenase-1 and -2 in human sporadic amyotrophic lateral sclerosis.
Abstract Prostaglandins (PGs) are critical mediators of physiologic processes and inflammation. They are produced by two different isoforms of the cyclooxygenase (COX) enzyme, namely COX-1 and COX-2. In particular COX-2 was demonstrated to be crucial for PG-synthesis in inflammation. Recently, inhibition of COX-2 was shown to prevent the loss of motor neurons in a model of amyotrophic lateral sclerosis (ALS). Furthermore, spinal COX-2 expression was shown to be increased in transgenic mice that produce an ALS-like syndrome. Therefore, we investigated the expression of COX-1 and COX-2 in the spinal cord of seven human sporadic ALS patients by means of immunohistochemistry. Specimens from seven patients without any neurological disease served as controls. COX-2 expression was dramatically increased in the spinal cord of patients with ALS. Its protein was found in motor neurons, interneurons and glial cells. Statistical analysis showed a significantly higher expression of COX-2 in ALS for both neurons and glia. In contrast, COX-1 expression was predominantly confined to microglia and no apparent difference was detected between controls and ALS. In addition, we studied the concentration of prostaglandin E2 (PG E2) as a marker for COX activity in the cerebrospinal fluid of nine patients diagnosed for ALS and compared the results with those from nine patients without motor neuron disease. PG E2 levels were markedly increased in ALS cases (45.8 +/- 35.1 pg/mL) compared to the non-ALS specimens (15.8 +/- 3.7 pg/mL). The results of our study corroborate a potential role for COX-2 in the pathogenesis of motor neuron death in ALS. Selective COX-2 inhibition might therefore offer a new possibility in the treatment of human ALS. However, to determine the exact role of COX-2 in human ALS will require further research. 6, Erlangen, Germany. maihoefner@physiologie1.uni-erlqngen.de

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Targets by SciMiner Summary

HUGO ID Symbol Target Name #Occur ActualStr
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)217COX | COX-2 | COX-2-positive | COX-immunoreactivity | COX-activity | cox 2 |
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)32COX-1 |
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))10SOD1 | mSOD1 | SOD | superoxide dismutase |
6018IL6interleukin 6 (interferon, beta 2)8IL-6 | il 6 |
5992IL1Binterleukin 1, beta7IL-1 | il 1 | IL-1beta |
4235GFAPglial fibrillary acidic protein6glial fibrillary acidic protein | GFAP |
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)6mac 1 | MAC-1 |
7794NFKB1nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (p105)1nf kappa b |
437ALPIalkaline phosphatase, intestinal1alkaline phosphatase |
10417RPS27Aribosomal protein S27a1ubiquitin |
2367CRPC-reactive protein, pentraxin-related1c reactive protein |

 

Targets by SciMiner Full list

HUGO ID Symbol Name ActualStr Score FlankingText
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2are produced by two different isoforms of the cyclooxygenase (COX) COX enzyme namely COX-1 and COX-2
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4two different isoforms of the cyclooxygenase (COX) COX enzyme namely COX-1 and COX-2
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4two different isoforms of the cyclooxygenase (COX) COX enzyme namely COX-1 and COX-2
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4isoforms of the cyclooxygenase (COX) COX enzyme namely COX-1 and COX-2
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4In particular COX-2 was demonstrated to be crucial for PG-synthesis in inflammation
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Recently inhibition of COX-2 was shown to prevent the loss of motor neurons in
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Furthermore spinal COX-2 expression was shown to be increased in transgenic mice that
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Therefore we investigated the expression of COX-1 and COX-2 in the spinal cord of seven human sporadic
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore we investigated the expression of COX-1 and COX-2 in the spinal cord of seven human sporadic
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore we investigated the expression of COX-1 and COX-2 in the spinal cord of seven human sporadic ALS patients
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4COX-2 expression was dramatically increased in the spinal cord of patients
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Statistical analysis showed a significantly higher expression of COX-2 in ALS for both neurons and glia
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4In contrast COX-1 expression was predominantly confined to microglia and no apparent difference
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2prostaglandin E2 (PG PG E 2 as a marker for COX activity in the cerebrospinal fluid of nine patients diagnosed for
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4The results of our study corroborate a potential role for COX-2 in the pathogenesis of motor neuron death in ALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Selective COX-2 inhibition might therefore offer a new possibility in the treatment
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4However to determine the exact role of COX-2 in human ALS will require further research
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))SOD12.9gene encoding for the Cu/Zn Cu Zn superoxide dismutase (SOD1) SOD1 account for a familial form of ALS linked to chromosome
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))SOD12.9Transgenic mice expressing mutant SOD1 develop a phenotype that mimics the clinical and pathological characteristics
5992IL1Binterleukin 1, betaIL-1beta1.5Levels of IL-1beta IL-6 and tumour necrosis factor-alpha were found to be elevated
6018IL6interleukin 6 (interferon, beta 2)IL-61.0Levels of IL-1beta IL-6 and tumour necrosis factor-alpha were found to be elevated in
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Very recently the proinflammatory enzyme cyclooxygenase-2 (COX-2) COX-2 was reported to be highly expressed in the spinal cord
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2Currently two COX isoforms are known namely COX-1 and COX-2 ( Vane et
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Currently two COX isoforms are known namely COX-1 and COX-2 ( Vane et al . 1998
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Currently two COX isoforms are known namely COX-1 and COX-2 ( Vane et al . 1998
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Currently two COX isoforms are known namely COX-1 and COX-2 ( Vane et al . 1998
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Whereas COX-1 mainly subserves _amp_#8216 house-keeping_amp_#8217 functions COX-2 is the product of
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Whereas COX-1 mainly subserves _amp_#8216 house-keeping_amp_#8217 functions COX-2 is the product of an _amp_#8216 immediate-early gene_amp_#8217 that is
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Under normal conditions COX-2 expression is highly restricted to distinct organ systems including the
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4and the eye ( Maihofner et al . 2001 but COX-2 expression can be dramatically increased in various tissues following the
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2Recently one of us demonstrated the constitutive expression of both COX isoforms in the spinal cord of rodents and a dramatic
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4spinal cord of rodents and a dramatic induction of spinal COX-2 protein following peripheral nociceptive stimulation ( Maihofner et al .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Very recently inhibition of COX-2 was protective in a glutamate-mediated in vitro model of sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4et al . (2001 2001 described the immunohistochemical distribution of COX-2 in the spinal cord of mSOD1 mice over the progression
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))mSOD13.2the immunohistochemical distribution of COX-2 in the spinal cord of mSOD1 mice over the progression of the disease
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4post mortem samples of ALS cases as a marker for COX-2 activity
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2However the cellular localization and expression of both COX isoforms in human sALS remains unclear
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Therefore we compared the spinal expression of COX-1 and COX-2 in sALS and control patients by means of
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore we compared the spinal expression of COX-1 and COX-2 in sALS and control patients by means of
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore we compared the spinal expression of COX-1 and COX-2 in sALS and control patients by means of immunohistochemistry
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4We here demonstrate a high expression of COX-2 protein in spinal cord specimens of human sALS cases
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4COX-1 expression did not differ between sALS and control tissues
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4The increased expression of COX-2 was corroborated by a significantly higher concentration of PG E
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Briefly goat polyclonal antisera raised against human COX-1 and COX-2 protein (Santa Santa Cruz Biotechnology Santa Cruz CA
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Briefly goat polyclonal antisera raised against human COX-1 and COX-2 protein (Santa Santa Cruz Biotechnology Santa Cruz CA
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Briefly goat polyclonal antisera raised against human COX-1 and COX-2 protein (Santa Santa Cruz Biotechnology Santa Cruz CA were employed
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4To further differentiate the cellular distributions of COX-1 and COX-2 proteins a double staining procedure using a mouse
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4To further differentiate the cellular distributions of COX-1 and COX-2 proteins a double staining procedure using a mouse
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4To further differentiate the cellular distributions of COX-1 and COX-2 proteins a double staining procedure using a mouse monoclonal antihuman
4235GFAPglial fibrillary acidic proteinGFAP2.5antibody to glial fibrillary acidic protein to label astrocytes (GFAP; GFAP DAKO Glostrup Denmark and a mouse monoclonal antihuman antibody to
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)MAC-11.0monoclonal antihuman antibody to macrophage antigen complex-1 labelling microglia (MAC-1, MAC-1 Serotec Raleigh NC was used
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Specimens were coded and assessed observer-blinded for presence of COX-1 and COX-2 immunoreactivity (IR) IR
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Specimens were coded and assessed observer-blinded for presence of COX-1 and COX-2 immunoreactivity (IR) IR
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4were coded and assessed observer-blinded for presence of COX-1 and COX-2 immunoreactivity (IR) IR
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Membranes were probed with polyclonal goat antihuman COX-1 or COX-2 serum (diluted diluted 1 1000 followed by a
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Membranes were probed with polyclonal goat antihuman COX-1 or COX-2 serum (diluted diluted 1 1000 followed by a horseradish peroxidase
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4immunohistochemistry we performed Western blotting experiments for the detection of COX-1 and COX-2 protein with spinal protein extracts from human donors
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4immunohistochemistry we performed Western blotting experiments for the detection of COX-1 and COX-2 protein with spinal protein extracts from human donors
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4performed Western blotting experiments for the detection of COX-1 and COX-2 protein with spinal protein extracts from human donors with no
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4cords (segments segments L1-L5 left lane run alongside purified sheep COX-1 protein (right right lane in a Western blot that has
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Western blot that has been incubated with the same polyclonal COX-1 antibody as that used for immunohistochemistry
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.475 kDa consistent with the reported molecular mass of human COX-1 protein in the literature (74 74 kDa ( Vane et
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4the results of a similar experiment for the detection of COX-2 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4lane from human spinal cords were separated alongside purified sheep COX-2 protein (right right lane by SDS-PAGE blotted onto nitrocellulose membranes
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4SDS-PAGE blotted onto nitrocellulose membranes and incubated with the same COX-2 antibody as that used for immunohistochemistry
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Unpurified COX-2 proteins derived from either cell lysates or tissues typically produce
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4single band at 75 kDa was detected for the purified COX-2 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2To exclude potential crossreactions between the antibodies for the two COX isoforms we determined the specifity with purified COX-1 and COX-2
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4the two COX isoforms we determined the specifity with purified COX-1 and COX-2 protein by Western blotting
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4the two COX isoforms we determined the specifity with purified COX-1 and COX-2 protein by Western blotting
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4COX isoforms we determined the specifity with purified COX-1 and COX-2 protein by Western blotting
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Figure 1C shows that the goat antihuman polyclonal COX-1 antibody exclusively detected COX-1 protein and not COX-2 protein
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4shows that the goat antihuman polyclonal COX-1 antibody exclusively detected COX-1 protein and not COX-2 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4antihuman polyclonal COX-1 antibody exclusively detected COX-1 protein and not COX-2 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4from a similar experiment in which the goat antihuman polyclonal COX-2 antibody detected only COX-2 protein and not COX-1 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4in which the goat antihuman polyclonal COX-2 antibody detected only COX-2 protein and not COX-1 protein
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4antihuman polyclonal COX-2 antibody detected only COX-2 protein and not COX-1 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-immunoreactivity0.0COX-immunoreactivity
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4only a few motor neurons and interneurons were immunoreactive for COX-2 protein ( Fig 2B
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-2-positive0.0number of neurons with COX-2-IR was counted the percentage of COX-2-positive motor neurons and interneurons was found to be significantly increased
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Furthermore expression of COX-2 was found to be enhanced in sALS cases compared to
4235GFAPglial fibrillary acidic proteinGFAP2.5cells counterstaining with an antibody raised against the astrocyte marker GFAP was performed
4235GFAPglial fibrillary acidic proteinGFAP2.5As demonstrated by the coexpression with GFAP glial COX-2 expression was predominantly found in astrocytes ( Fig
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4As demonstrated by the coexpression with GFAP glial COX-2 expression was predominantly found in astrocytes ( Fig 2H
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)MAC-11.0microglia as shown by its coexpression with the microglia marker MAC-1 ( Fig 2I
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Overall the glial expression of COX-2 was significantly increased in sALS cases (0.71 0.71 _amp_plusmn 0.5
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Expression of COX-1 protein was predominantly confined to some small cells with the
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4This could also be demonstrated by the costaining of COX-1 with the microglial marker MAC-1 ( Fig 2L
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)MAC-11.0demonstrated by the costaining of COX-1 with the microglial marker MAC-1 ( Fig 2L
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4reduction of neurons in the spinal cord the percentage of COX-2 expressing motor neurons and interneurons was increased in sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Secondly glial expression of COX-2 was enhanced in sALS
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Thirdly COX-1 expression was predominately confined to glial cells
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4COX-2 overexpression in sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Expression of COX-2 in human diseases is a topic of considerable interest with
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4of considerable interest with regard to the recent development of COX-2 selective inhibitors ( Hawkey 1999
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4( Drachman _amp_ Rothstein 2000 demonstrated a beneficial effect of COX-2 inhibition in an in vitro model while Almer et al
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4(2001 2001 demonstrated a role for COX-2 in mutant SOD1 mice and Yasojima et al
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))SOD12.9(2001 2001 demonstrated a role for COX-2 in mutant SOD1 mice and Yasojima et al
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4(2001 2001 showed up-regulation of COX-2 mRNA in spinal cord specimens of sALS cases
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Recently two animal studies demonstrated a beneficial effect of selective COX-2 inhibition in mouse models of ALS ( Drachman et al
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4However the cellular origin of COX-2 up-regulation in human sALS has so far not been delineated
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Furthermore no study has yet investigated the cellular expression of COX-1 in sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4we provide immunohistochemical evidence for a dramatic increase in spinal COX-2 expression in motor neurons interneurons and glial cells in sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4COX-2 was also observed in motor neurons of control specimens the
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4This agrees with reports on the constitutive expression of COX-2 in the spinal cord of rodents and the localization of
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4in the spinal cord of rodents and the localization of COX-2 protein determined electron microscopically ( Maihofner et al . 2000b
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4In normal conditions COX-2 derived PGs are assumed to play homeostatic functions ( Beiche
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Nevertheless the reason for the dramatic COX-2 overexpression in sALS is unknown
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4In pain models COX-2 protein was induced following glutamatergic stimulation ( Maihofner et al
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4NF-kappa B NF-kappa B in particular is crucial for COX-2 induction in several cell types ( Vane et al .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Cytokines could also contribute to high expression of COX-2 in sALS particularly IL-1beta and IL-6
5992IL1Binterleukin 1, betaIL-1beta1.5also contribute to high expression of COX-2 in sALS particularly IL-1beta and IL-6
6018IL6interleukin 6 (interferon, beta 2)IL-61.0to high expression of COX-2 in sALS particularly IL-1beta and IL-6
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Both are known inducers of COX-2 expression and their concentrations are elevated in the CSF of
5992IL1Binterleukin 1, betaIL-1beta1.5Interestingly inhibition of IL-1beta also attenuates the loss of motor neurons in mSOD1 mice
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))mSOD13.2of IL-1beta also attenuates the loss of motor neurons in mSOD1 mice ( Friedlander et al . 1997
5992IL1Binterleukin 1, betaIL-1beta1.5IL-1beta and IL-6 are assumed to derive from glia
6018IL6interleukin 6 (interferon, beta 2)IL-61.0IL-1beta and IL-6 are assumed to derive from glia
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Consistently we demonstrated a significantly higher expression of COX-2 in glial cells
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))mSOD13.2In agreement with a study in transgenic mSOD1 mice ( Almer et al . 2001 COX-2 was found
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4in transgenic mSOD1 mice ( Almer et al . 2001 COX-2 was found to be predominantly expressed in astrocytes and not
5992IL1Binterleukin 1, betaIL-1beta1.5This might be explained by the capability of IL-1beta to induce COX-2 in astrocytes but not in microglia cells
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4might be explained by the capability of IL-1beta to induce COX-2 in astrocytes but not in microglia cells ( Vane et
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4COX-2 expression in the astroglia of rodents has been shown previously
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4reduction in the number of interneurons in our sALS cases COX-2 expression was also found to be increased in this cell
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2match those of a previous study investigating the expression of COX proteins in mSOD1 mice
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))mSOD13.2a previous study investigating the expression of COX proteins in mSOD1 mice
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4(2001 2001 also found that COX-2 was present in motor neurons and predominantly in astroglia although
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4neurons and predominantly in astroglia although no apparent differences in COX-1 expression were seen between mSOD1 mice and controls
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))mSOD13.2although no apparent differences in COX-1 expression were seen between mSOD1 mice and controls
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4that misclassification of neurons led to the high expression of COX-2 protein in sALS presented here
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4. 2003 and were shown to be specific for the COX-1 and COX-2 protein respectively
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4. 2003 and were shown to be specific for the COX-1 and COX-2 protein respectively
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4and were shown to be specific for the COX-1 and COX-2 protein respectively
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2of the antibodies and their capability to detect the respective COX isoforms in human spinal cord extracts by Western blotting
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4have intentionally focused on the in situ expression of both COX-1 and COX-2 proteins in human ALS spinal cords
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4have intentionally focused on the in situ expression of both COX-1 and COX-2 proteins in human ALS spinal cords
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4focused on the in situ expression of both COX-1 and COX-2 proteins in human ALS spinal cords
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2Therefore this study provides necessary data regarding the expression of COX isoforms in human ALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-activity0.0Therefore measurement of PG-E 2 as a marker for COX-activity may serve as an additional independent method
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4with the results of two recent animal studies where selective COX-2 inhibition was found to be protective against motor neuron death
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4A role for COX-2 in the pathogenesis of sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4What is the significance of COX-2 overexpression in ALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4There are several lines of evidence that COX-2 might play a role in the pathogenesis of ALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Firstly application of a selective COX-2 inhibitor in an in vitro organotypic model of ALS protected
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4in two in vivo studies where application of a selective COX-2 inhibitor protected against motor neuron degeneration and prolonged survival in
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Thirdly based on the action of PGs COX-2 could promote inflammatory processes in ALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore COX-2 derived PGs could play a role in glutamate excitotoxicity which
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4potentiation of kainic acid induced excitotoxicity in transgenic mice overexpressing COX-2 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Fourthly COX-2 may also be involved in the production of reactive oxygen
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2COX enzymes are bifunctional proteins
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2The peroxidase activity of COX is nonspecific and can reduce several different hydroperoxides while co-oxidizing
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Finally COX-2 appears to be involved in neuronal cell cycle regulation
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4kinase (INK) INK p18INK4 is a downstream target of neuronal COX-2 expression p18INK4 inhibits CDK 4 6 which is in turn
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore COX-2 inhibition might attenuate apoptotic damage in neurodegenerative diseases
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4However the specifity of COX-2 expression for ALS has to be questioned
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Increased COX-2 levels were also shown for Alzheimer's disease Parkinson's disease epilepsy
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Therefore we would explicitly like to state that COX-2 expression seems to be a common endpoint in neurodegeneration rather
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Furthermore there is the possibility that COX-2 expression might be protective and antiapoptotic
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4of us was able to show that the expression of COX-2 in CRC parallels the expression of IL-1 beta and IL-6
5992IL1Binterleukin 1, betaIL-11.5the expression of COX-2 in CRC parallels the expression of IL-1 beta and IL-6 in CRC ( Maihofner et al .
6018IL6interleukin 6 (interferon, beta 2)IL-61.0COX-2 in CRC parallels the expression of IL-1 beta and IL-6 in CRC ( Maihofner et al . 2003
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4In this context COX-2 is thought to be actively involved in carcinogenesis and antiapoptosis
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4by the immunohistochemical results presented here the exact role of COX-2 in the pathogenesis of human ALS still remains to be
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4North-east ALS consortium to test a potential benefit of selective COX-2 inhibition in human ALS
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Expression of COX-1 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4In contrast to the COX-2 protein COX-1 expression was predominantly found in microglia cells and
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4In contrast to the COX-2 protein COX-1 expression was predominantly found in microglia cells and showed no
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4Expression of COX-1 in microglia is consistent with our previous animal study and
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4(2001 2001 who reported a similar content of COX-1 mRNA in sALS and controls
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4The unaltered expression of COX-1 in sALS implies that there is no or only a
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Alzheimer disease ( Pasinetti _amp_ Aisen 1998 where COX-2 was found to be the pivotal isoform involved in the
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2for the first time the in situ expression of both COX proteins in the spinal cords of human sALS specimens
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Expression of COX-2 protein was markedly increased in the motoneurons interneurons and glial
9604PTGS1prostaglandin-endoperoxide synthase 1 (prostaglandin G/H synthase and cyclooxygenase)COX-15.4was detected between controls and sALS with regard to the COX-1 protein
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4Based on the beneficial effects of selective COX-2 inhibition in models of ALS the results of this and
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX-213.4rationale for clinical investigations on potential positive effects of selective COX-2 inhibitors in human sALS
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)COX2.2ALS amyotrophic lateral sclerosis COX cyclooxygenase CSF cerebrospinal fluid GFAP glial fibrillary acidic protein IR
4235GFAPglial fibrillary acidic proteinGFAP2.5ALS amyotrophic lateral sclerosis COX cyclooxygenase CSF cerebrospinal fluid GFAP glial fibrillary acidic protein IR immunoreactivity PG prostaglandins SOD superoxide
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))SOD2.9fluid GFAP glial fibrillary acidic protein IR immunoreactivity PG prostaglandins SOD superoxide dismutase
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0they are produced by two different isoforms of the cyclooxygenase cox enzyme namely cox 1 and cox 2.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in particular cox 2 was demonstrated to be crucial for pg synthesis in inflammation.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0recently inhibition of cox 2 was shown to prevent the loss of motor neurons in a model of amyotrophic lateral sclerosis als .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0furthermore spinal cox 2 expression was shown to be increased in transgenic mice that produce an als like syndrome.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore we investigated the expression of cox 1 and cox 2 in the spinal cord of seven human sporadic als patients by means of immunohistochemistry.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0cox 2 expression was dramatically increased in the spinal cord of patients with als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0statistical analysis showed a significantly higher expression of cox 2 in als for both neurons and glia.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0the results of our study corroborate a potential role for cox 2 in the pathogenesis of motor neuron death in als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0selective cox 2 inhibition might therefore offer a new possibility in the treatment of human als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0however to determine the exact role of cox 2 in human als will require further research.
6018IL6interleukin 6 (interferon, beta 2)il 61.0levels of il 1beta il 6 and tumour necrosis factor alpha were found to be elevated in experimental and human sals ghezzi et al . 1998 ; sekizawa et al . 1998 ; li et al . 2000 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0very recently the proinflammatory enzyme cyclooxygenase 2 cox 2 was reported to be highly expressed in the spinal cord of msod1 mice almer et al . 2001 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0currently two cox isoforms are known namely cox 1 and cox 2 vane et al . 1998 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0whereas cox 1 mainly subserves _amp_#8216;house keeping_amp_#8217; functions cox 2 is the product of an _amp_#8216;immediate early gene_amp_#8217; that is rapidly inducible and tightly regulated vane et al . 1998 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0under normal conditions cox 2 expression is highly restricted to distinct organ systems including the kidney harris et al . 1994 and the eye maihofner et al . 2001 but cox 2 expression can be dramatically increased in various tissues following the initiation of transcription by activating factors including different proinflammatory cytokines arterial wall sheer forces or
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0recently one of us demonstrated the constitutive expression of both cox isoforms in the spinal cord of rodents and a dramatic induction of spinal cox 2 protein following peripheral nociceptive stimulation maihofner et al . 2000b .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0very recently inhibition of cox 2 was protective in a glutamate mediated in vitro model of sals drachman _amp_ rothstein 2000 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0almer et al . 2001 described the immunohistochemical distribution of cox 2 in the spinal cord of msod1 mice over the progression of the disease.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0furthermore they showed an increased prostaglandin e2 pg e 2 level in post mortem samples of als cases as a marker for cox 2 activity.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore we compared the spinal expression of cox 1 and cox 2 in sals and control patients by means of immunohistochemistry.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0we here demonstrate a high expression of cox 2 protein in spinal cord specimens of human sals cases.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0the increased expression of cox 2 was corroborated by a significantly higher concentration of pg e 2 in the cerebrospinal fluid of patients diagnosed with als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0briefly goat polyclonal antisera raised against human cox 1 and cox 2 protein santa cruz biotechnology santa cruz ca were employed.
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)mac 11.0nal antihuman antibody to glial fibrillary acidic protein to label astrocytes gfap; dako glostrup denmark and a mouse monoclonal antihuman antibody to macrophage antigen complex 1 labelling microglia mac 1 serotec raleigh nc was used.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0to further differentiate the cellular distributions of cox 1 and cox 2 proteins a double staining procedure using a mouse monoclonal antihuman antibody to glial fibrillary acidic protein to label astrocytes gfap; dako glostrup denmark and a mouse monoclonal antihuman an
4235GFAPglial fibrillary acidic proteinglial fibrillary acidic protein1.0to further differentiate the cellular distributions of cox 1 and cox 2 proteins a double staining procedure using a mouse monoclonal antihuman antibody to glial fibrillary acidic protein to label astrocytes gfap; dako glostrup denmark and a mouse monoclonal antihuman antibody to macrophage antigen complex 1 labelling microglia mac 1 serotec raleigh nc was used.
437ALPIalkaline phosphatase, intestinalalkaline phosphatase1.0the bound primary antibodies were visualized according to the streptavidin biotin complex sbc and alkaline phosphatase antialkaline phosphatase apaap methods alone or in combination with double staining procedures probst cousin et al . 2002 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0specimens were coded and assessed observer blinded for presence of cox 1 and cox 2 immunoreactivity ir .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0membranes were probed with polyclonal goat antihuman cox 1 or cox 2 serum diluted 1 : 1000 followed by a horseradish peroxidase hrp linked donkey antigoat igg secondary antibody diluted 1 : 1000; santa cruz biotechnology .
2367CRPC-reactive protein, pentraxin-relatedc reactive protein1.0a simultaneous infection was ruled out by routine laboratory assessment of c reactive protein leucocyte count and erythrocyte sedimentation rate.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0the normal spinal cord and specifity of the antibodies to assess the specifity of the antibodies used for immunohistochemistry we performed western blotting experiments for the detection of cox 1 and cox 2 protein with spinal protein extracts from human donors with no known neurological diseases two males age 65 and 78 years .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0figure 1b demonstrates the results of a similar experiment for the detection of cox 2 protein.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0protein extracts left lane from human spinal cords were separated alongside purified sheep cox 2 protein right lane by sds page blotted onto nitrocellulose membranes and incubated with the same cox 2 antibody as that used for immunohistochemistry.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0unpurified cox 2 proteins derived from either cell lysates or tissues typically produce a double or triple band in western blot analysis at 68 75 kda fig 1b left lane .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0a single band at 75 kda was detected for the purified cox 2 protein.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0to exclude potential crossreactions between the antibodies for the two cox isoforms we determined the specifity with purified cox 1 and cox 2 protein by western blotting.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0figure 1c shows that the goat antihuman polyclonal cox 1 antibody exclusively detected cox 1 protein and not cox 2 protein.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0figure 1d shows the results from a similar experiment in which the goat antihuman polyclonal cox 2 antibody detected only cox 2 protein and not cox 1 protein.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in control specimens only a few motor neurons and interneurons were immunoreactive for cox 2 protein fig 2b .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in most neurons cox 2 ir was absent fig 2c .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0the immunoreactive product could be clearly distinguished from lipofuscin like inclusions which had a different staining pattern compared to the reddish cox 2 ir fig 2c .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0when the number of neurons with cox 2 ir was counted the percentage of cox 2 positive motor neurons and interneurons was found to be significantly increased in sals patients compared to controls despite an overall reduction in the number of neurons 13.31 _amp_plusmn; 5 vs 37.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0furthermore expression of cox 2 was found to be enhanced in sals cases compared to controls in cells that showed no neuronal morphology i.e. cells of presumed glial origin fig 2f and g .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0as demonstrated by the coexpression with gfap glial cox 2 expression was predominantly found in astrocytes fig 2h .
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)mac 11.0however cox 2 ir was occasionally detected in smaller cells with the morphological characteristics of microglia as shown by its coexpression with the microglia marker mac 1 fig 2i .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0however cox 2 ir was occasionally detected in smaller cells with the morphological characteristics of microglia as shown by its coexpression with the microglia marker mac 1 fig 2i .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0overall the glial expression of cox 2 was significantly increased in sals cases 0.71 _amp_plusmn; 0.5 vs 2.3 _amp_plusmn; 0.5; wilcoxon signed ranked test p _lt_ 0.05; fig 5 .
6149ITGAMintegrin, alpha M (complement component 3 receptor 3 subunit)mac 11.0this could also be demonstrated by the costaining of cox 1 with the microglial marker mac 1 fig 2l .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0firstly despite a significant reduction of neurons in the spinal cord the percentage of cox 2 expressing motor neurons and interneurons was increased in sals.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0secondly glial expression of cox 2 was enhanced in sals.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0cox 2 overexpression in sals
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0expression of cox 2 in human diseases is a topic of considerable interest with regard to the recent development of cox 2 selective inhibitors hawkey 1999 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0regarding als drachman and rothstein drachman _amp_ rothstein 2000 demonstrated a beneficial effect of cox 2 inhibition in an in vitro model while almer et al .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0 2001 demonstrated a role for cox 2 in mutant sod1 mice and yasojima et al .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0 2001 showed up regulation of cox 2 mrna in spinal cord specimens of sals cases.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0recently two animal studies demonstrated a beneficial effect of selective cox 2 inhibition in mouse models of als drachman et al . 2002 ; pompl et al . 2003 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0however the cellular origin of cox 2 up regulation in human sals has so far not been delineated.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0here we provide immunohistochemical evidence for a dramatic increase in spinal cox 2 expression in motor neurons interneurons and glial cells in sals despite an overall reduction in the number of neurons compared to controls.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0cox 2 was also observed in motor neurons of control specimens the staining was accentuated in the perinuclear region.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0this agrees with reports on the constitutive expression of cox 2 in the spinal cord of rodents and the localization of cox 2 protein determined electron microscopically maihofner et al . 2000b ; maihofner et al . 2001 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in normal conditions cox 2 derived pgs are assumed to play homeostatic functions beiche et al . 1996 ; vane et al . 1998 ; maihofner et al . 2000b .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0nevertheless the reason for the dramatic cox 2 overexpression in sals is unknown.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0one potential explanation could be cox 2's interplay with glutamate levels of which are elevated in the csf of sals patients plaitakis et al . 1988 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in pain models cox 2 protein was induced following glutamatergic stimulation maihofner et al . 2000b .
7794NFKB1nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (p105)nf kappa b1.0nf kappa b. nf kappa b in particular is crucial for cox 2 induction in several cell types vane et al . 1998 and its activation has also been linked to neurodegeneration grilli _amp_ memo 1999 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0nf kappa b. nf kappa b in particular is crucial for cox 2 induction in several cell types vane et al . 1998 and its activation has also been linked to neurodegeneration grilli _amp_ memo 1999 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0cytokines could also contribute to high expression of cox 2 in sals particularly il 1beta and il 6.
6018IL6interleukin 6 (interferon, beta 2)il 61.0cytokines could also contribute to high expression of cox 2 in sals particularly il 1beta and il 6.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0both are known inducers of cox 2 expression and their concentrations are elevated in the csf of als cases sekizawa et al . 1998 ; vane et al . 1998 ; li et al . 2000 .
6018IL6interleukin 6 (interferon, beta 2)il 61.0il 1beta and il 6 are assumed to derive from glia.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0consistently we demonstrated a significantly higher expression of cox 2 in glial cells.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in agreement with a study in transgenic msod1 mice almer et al . 2001 cox 2 was found to be predominantly expressed in astrocytes and not microglia.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0this might be explained by the capability of il 1beta to induce cox 2 in astrocytes but not in microglia cells vane et al . 1998 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0cox 2 expression in the astroglia of rodents has been shown previously beiche et al . 1996 ; maihofner et al . 2000b .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0furthermore despite a significant reduction in the number of interneurons in our sals cases cox 2 expression was also found to be increased in this cell type.
10417RPS27Aribosomal protein S27aubiquitin1.0this is consistent with other studies showing an altered protein expression e.g. ubiquitin stephens et al . 2001 and an active involvement of interneurons in the pathology of als rowland _amp_ shneider 2001 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0 2001 also found that cox 2 was present in motor neurons and predominantly in astroglia although no apparent differences in cox 1 expression were seen between msod1 mice and controls.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore we do not think that misclassification of neurons led to the high expression of cox 2 protein in sals presented here.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0been used in previous studies maihofner et al . 2000b ; damm et al . 2001 ; maihofner et al . 2001 ; charalambous et al . 2003 ; maihofner et al . 2003 and were shown to be specific for the cox 1 and cox 2 protein respectively.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0we have intentionally focused on the in situ expression of both cox 1 and cox 2 proteins in human als spinal cords.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0increased pg e 2 levels are also in agreement with the results of two recent animal studies where selective cox 2 inhibition was found to be protective against motor neuron death drachman et al . 2002 ; pompl et al . 2003 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0a role for cox 2 in the pathogenesis of sals?
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0what is the significance of cox 2 overexpression in als?
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0there are several lines of evidence that cox 2 might play a role in the pathogenesis of als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0firstly application of a selective cox 2 inhibitor in an in vitro organotypic model of als protected against the loss of motor neurons in this system drachman _amp_ rothstein 2000 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0secondly this finding was corroborated in two in vivo studies where application of a selective cox 2 inhibitor protected against motor neuron degeneration and prolonged survival in transgenic mouse models of als drachman et al . 2002 ; pompl et al . 2003 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0thirdly based on the action of pgs cox 2 could promote inflammatory processes in als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore cox 2 derived pgs could play a role in glutamate excitotoxicity which is postulated to occur in als rowland _amp_ shneider 2001 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0this is also in line with the study of kelley and colleagues kelley et al . 1999 demonstrating a potentiation of kainic acid induced excitotoxicity in transgenic mice overexpressing cox 2 protein.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0fourthly cox 2 may also be involved in the production of reactive oxygen species and oxidative stress vane et al . 1998 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0finally cox 2 appears to be involved in neuronal cell cycle regulation.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0 2002 showed that the expression of the endogenous cell cycle dependent kinase cdk inhibitor inhibitor kinase ink p18ink4 is a downstream target of neuronal cox 2 expression. p18ink4 inhibits cdk 4 6 which is in turn a key player in the regulation of g1 progression zindy et al . 1997 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore cox 2 inhibition might attenuate apoptotic damage in neurodegenerative diseases.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0however the specifity of cox 2 expression for als has to be questioned.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0increased cox 2 levels were also shown for alzheimer's disease parkinson's disease epilepsy and even cerebral infarction pasinetti _amp_ aisen 1998 ; vane et al . 1998 ; scali et al . 2000 ; kunz _amp_ oliw 2001 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore we would explicitly like to state that cox 2 expression seems to be a common endpoint in neurodegeneration rather than the actual cause of als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0furthermore there is the possibility that cox 2 expression might be protective and antiapoptotic.
5992IL1Binterleukin 1, betail 11.0recently one of us was able to show that the expression of cox 2 in crc parallels the expression of il 1 beta and il 6 in crc maihofner et al . 2003 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0recently one of us was able to show that the expression of cox 2 in crc parallels the expression of il 1 beta and il 6 in crc maihofner et al . 2003 .
6018IL6interleukin 6 (interferon, beta 2)il 61.0recently one of us was able to show that the expression of cox 2 in crc parallels the expression of il 1 beta and il 6 in crc maihofner et al . 2003 .
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in this context cox 2 is thought to be actively involved in carcinogenesis and antiapoptosis.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0therefore although the animal findings are corroborated by the immunohistochemical results presented here the exact role of cox 2 in the pathogenesis of human als still remains to be elucidated.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0clinical trials are underway in the united states north east als consortium to test a potential benefit of selective cox 2 inhibition in human als.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0in contrast to the cox 2 protein cox 1 expression was predominantly found in microglia cells and showed no significant difference between sals and controls.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0alzheimer disease pasinetti _amp_ aisen 1998 where cox 2 was found to be the pivotal isoform involved in the pathology.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0expression of cox 2 protein was markedly increased in the motoneurons interneurons and glial cells of sals cases compared to controls.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0based on the beneficial effects of selective cox 2 inhibition in models of als the results of this and other studies might give a rationale for clinical investigations on potential positive effects of selective cox 2 inhibitors in human sals.
9605PTGS2prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase)cox 21.0 inhibition in models of als the results of this and other studies might give a rationale for clinical investigations on potential positive effects of selective cox 2 inhibitors in human sals.
4235GFAPglial fibrillary acidic proteinglial fibrillary acidic protein1.0als amyotrophic lateral sclerosis; cox cyclooxygenase; csf cerebrospinal fluid; gfap glial fibrillary acidic protein; ir immunoreactivity; pg prostaglandins; sod superoxide dismutase.
11179SOD1superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult))superoxide dismutase1.0als amyotrophic lateral sclerosis; cox cyclooxygenase; csf cerebrospinal fluid; gfap glial fibrillary acidic protein; ir immunoreactivity; pg prostaglandins; sod superoxide dismutase.