Identification of alternate transcripts of neuronal nitric oxide synthase in the mouse retina
Autor: | William D. Eldred, Thomas J. Giove, Monika M. Deshpande |
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Rok vydání: | 2009 |
Předmět: |
Male
Cell type Blotting Western Alpha (ethology) Nitric Oxide Synthase Type I Biology Nitric Oxide Retina Nitric oxide Mice Cellular and Molecular Neuroscience chemistry.chemical_compound medicine Animals Protein Isoforms Beta (finance) reproductive and urinary physiology Reverse Transcriptase Polymerase Chain Reaction Alternative splicing musculoskeletal system Immunohistochemistry Cell biology Mice Inbred C57BL body regions Blot Alternative Splicing medicine.anatomical_structure nervous system chemistry cardiovascular system Neuroscience |
Zdroj: | Journal of Neuroscience Research. 87:3134-3142 |
ISSN: | 1097-4547 0360-4012 |
DOI: | 10.1002/jnr.22133 |
Popis: | Nitric oxide (NO) is a major signaling molecule in the retina and CNS, with physiological roles in every cell type in the retina. Previous work shows that neuronal nitric oxide synthase (nNOS) is an important source of NO in the vertebrate retina. There are distinct, active alternative transcripts of nNOS observed in many tissues, including testes and brain, that may differ in both localization and enzyme kinetics. The present study characterized nNOS and the NO production from nNOS in the mouse retina in terms of its alternate transcripts, namely, nNOS alpha, nNOS beta, and nNOS gamma. We examined both basal and light-stimulated NO production as imaged using the NO-sensitive dye 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate-FM (DAF-FM), and we compared the NO production with the immunocytochemical localization of nNOS using antisera that recognize nNOS alpha/beta or nNOS alpha/beta/gamma. Western blots suggested the presence of NOS alpha/gamma protein in retina, but not nNOS beta, and we confirmed this at the message level by using a combination of RT-PCR and quantitative real-time PCR. Our findings indicated that the primary source of NO in the mammalian retina is nNOS alpha and that nNOS gamma may contribute to NO production as well. |
Databáze: | OpenAIRE |
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