title: The Presence of the Neuronal Nitric Oxide Synthase Isoform in the Intervertebral Disk.
abstract: Intervertebral disk degeneration is a progressive and debilitating disease with multifactorial causes. Nitric oxide (NO) might contribute to the cell death pathway. We evaluated the presence of the constitutive form of the neuronal NOS (nNOS) in both health and degenerated intervertebral disk through qPCR and immunohistochemistry. We also analyzed the potential role of nNOS modulation in the tail needle puncture model of intervertebral disk degeneration. Male Wistar rats were submitted to percutaneous disk puncture with a 21-gauge needle of coccygeal vertebras. The selective nNOS pharmacological inhibitor N(ω)-propyl-L-arginine (NPLA) or a nNOS-target siRNA (siRNAnNOShum_4400) was injected immediately after the intervertebral disk puncture with a 30-gauge needle. Signs of disk degeneration were analyzed by in vivo magnetic resonance imaging and histological score. We found that intact intervertebral disks express low levels of nNOS mRNA. Disk injury caused a 4 fold increase in nNOS mRNA content at 5 h post disk lesion. However, NPLA or nNOS-target siRNA slight mitigate the intervertebral disk degenerative progress. Our data show evidence of the nNOS presence in the intervertebral disk and its upregulation during degeneration. Further studies would disclose the nNOS role and its potential therapeutical value in the intervertebral disk degeneration.
authors.affiliation: Department of Morphology, Physiology and Basic Pathology, Dental School, University of São Paulo (USP), Ribeirão Prêto, SP, 14049-904, Brazil.
authors.forename: Vitor
authors.initials: V
authors.lastname: Castania
authors.affiliation: Department of Morphology, Physiology and Basic Pathology, Dental School, University of São Paulo (USP), Ribeirão Prêto, SP, 14049-904, Brazil.|Center for Interdisciplinary Research on Applied Neurosciences (NAPNA), University of São Paulo (USP), São Paulo, Brazil.
authors.forename: Ana Carolina
authors.initials: AC
authors.lastname: Issy
authors.affiliation: Department of Morphology, Physiology and Basic Pathology, Dental School, University of São Paulo (USP), Ribeirão Prêto, SP, 14049-904, Brazil.
authors.forename: João Walter
authors.initials: JW
authors.lastname: Silveira
authors.affiliation: Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.|Center for Interdisciplinary Research on Applied Neurosciences (NAPNA), University of São Paulo (USP), São Paulo, Brazil.
authors.forename: Frederico Rogério
authors.initials: FR
authors.lastname: Ferreira
authors.affiliation: Technology for Gene Therapy Laboratory, University of Brasilia - UnB/FAV, Brasília, DF, Brazil.
authors.forename: Simoneide S
authors.initials: SS
authors.lastname: Titze-de-Almeida
authors.affiliation: Technology for Gene Therapy Laboratory, University of Brasilia - UnB/FAV, Brasília, DF, Brazil.
authors.forename: Fernando F B
authors.initials: FF
authors.lastname: Resende
authors.affiliation: Department of Morphology, Physiology and Basic Pathology, Dental School, University of São Paulo (USP), Ribeirão Prêto, SP, 14049-904, Brazil.
authors.forename: Nádia Rubia
authors.initials: NR
authors.lastname: Ferreira
authors.affiliation: Technology for Gene Therapy Laboratory, University of Brasilia - UnB/FAV, Brasília, DF, Brazil.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Titze-de-Almeida
authors.affiliation: Department of Biomechanics, Medicine and Rehabilitation of the Locomotor Apparatus, School of Medicine, University of São Paulo (USP), Ribeirão Prêto, Brazil.
authors.forename: Helton L A
authors.initials: HL
authors.lastname: Defino
authors.affiliation: Department of Morphology, Physiology and Basic Pathology, Dental School, University of São Paulo (USP), Ribeirão Prêto, SP, 14049-904, Brazil. eadelbel@forp.usp.br.|Center for Interdisciplinary Research on Applied Neurosciences (NAPNA), University of São Paulo (USP), São Paulo, Brazil. eadelbel@forp.usp.br.
authors.forename: Elaine
authors.initials: E
authors.lastname: Del Bel
mesh_terms: D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D055785:Gene Knockdown Techniques; D006801:Humans; D007150:Immunohistochemistry; D007403:Intervertebral Disc / Q000000981:diagnostic imaging / Q000201:enzymology* / Q000473:pathology; D055959:Intervertebral Disc Degeneration / Q000000981:diagnostic imaging / Q000201:enzymology* / Q000473:pathology; D008279:Magnetic Resonance Imaging; D008297:Male; D052248:Nitric Oxide Synthase Type I / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D017208:Rats, Wistar; D012445:Sacrococcygeal Region
chemical_list: D004791:Enzyme Inhibitors; C109599:N(omega)-propylarginine; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D001120:Arginine; C496320:NOS1 protein, human; D052248:Nitric Oxide Synthase Type I; C496322:Nos1 protein, rat
keywords: Animal model; Intervertebral disk degeneration; Nitric oxide; nNOS; siRNA