title: TRPV1 receptors are involved in protein nitration and Müller cell reaction in the acutely axotomized rat retina.
abstract: We report here the protein expression of TRPV1 receptor in axotomized rat retinas and its possible participation in mechanisms involved in retinal ganglion cell (RGC) death. Adult rats were subjected to unilateral, intraorbital axotomy of the optic nerve, and the retinal tissue was removed for further processing. TRPV1 total protein expression decreased progressively after optic nerve transection, reaching 66.2% of control values 21 days after axotomy. The number of cells labeled for TRPV1 in the remnant GCL decreased after 21 days post-lesion (to 63%). Fluoro-Jade B staining demonstrated that the activation of TRPV1 in acutely-lesioned eyes elicited more intense neuronal degeneration in the GCL and in the inner nuclear layer than in sham-operated retinas. A single intraocular injection of capsazepine (100 μM), a TRPV1 antagonist, 5 days after optic nerve lesion, decreased the number of GFAP-expressing Müller cells (72.5% of control values) and also decreased protein nitration in the retinal vitreal margin (75.7% of control values), but did not affect lipid peroxidation. Furthermore, retinal explants were treated with capsaicin (100 μM), and remarkable protein nitration was then present, which was reduced by blockers of the constitutive and inducible nitric oxide synthases (7-NI and aminoguanidine, respectively). TRPV1 activation also increased GFAP expression, which was reverted by both TRPV1 antagonism with capsazepine and by 7-NI and aminoguanidine. Given that Müller cells do not express TRPV1, we suppose that the increased GFAP expression in these cells might be elicited by TRPV1 activation and by its indirect effect upon nitric oxide overproduction and peroxynitrite formation. We incubated Fluorogold pre-labeled retinal explants in the presence of capsazepine (1 μM) during 48 h. The numbers of surviving RGCs stained with fluorogold and the numbers of apoptotic cells in the GCL detected with TUNEL were similar in lesioned and control retinas. We conclude that TRPV1 receptor expression decreased after optic nerve injury due to death of TRPV1-containing cells. Furthermore, these data indicate that TRPV1 might be involved in intrinsic protein nitration and Müller cell reaction observed after optic nerve injury.
authors.affiliation: Department of Physiology & Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP 05508-900, Brazil. mauroleonelli@yahoo.com.br
authors.forename: Mauro
authors.initials: M
authors.lastname: Leonelli
authors.forename: Daniel O
authors.initials: DO
authors.lastname: Martins
authors.forename: Luiz R G
authors.initials: LR
authors.lastname: Britto
mesh_terms: D000818:Animals; D019771:Axotomy; D002211:Capsaicin / Q000031:analogs & derivatives / Q000494:pharmacology; D002452:Cell Count; D005452:Fluoresceins; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D015227:Lipid Peroxidation; D009457:Neuroglia / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D015538:Nitrosation; D009900:Optic Nerve / Q000502:physiology*; D009930:Organic Chemicals; D051381:Rats; D017208:Rats, Wistar; D012165:Retinal Ganglion Cells / Q000378:metabolism* / Q000473:pathology; D050916:TRPV Cation Channels / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology*
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D009930:Organic Chemicals; D050916:TRPV Cation Channels; C494484:TRPV1 receptor; C493922:Trpv1 protein, rat; C435731:fluoro jade; D009569:Nitric Oxide; C071423:capsazepine; D002211:Capsaicin