Endothelial and neuronal nitric oxide synthases are present in the suprachiasmatic nuclei of Syrian hamsters and rats.
Nitric oxide (NO) is involved in the transmission of light information to suprachiasmatic nuclei (SCN). By immunocytochemistry, we showed that both neuronal and endothelial NO synthase isoforms (nNOS and eNOS) were present in the SCN of rats and hamsters. nNOS-immunoreactive neurons were located mainly around the SCN with only a few nNOS neurons within the nucleus. By double-label immunocytochemistry, we also found, within the population of SCN glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes, a subpopulation of eNOS-immunoreactive astrocytes. Using Western blot analysis, we detected in SCN protein extracts eNOS and nNOS proteins having the expected 140 and 150 kDa molecular weights, respectively. By in situ hybridization of a 2.4-kb murine eNOS probe, mRNA for eNOS was located in the SCN of rats and hamsters. The transcript was further identified by detection of a RT-PCR product of the predicted size, after amplification of total RNA with primers specific for eNOS. In the SCN and cerebellum, the size of the mRNA for nNOS, detected with a rat probe on Northern blot, was approximately 10.5 kb, corresponding to that previously published. In the same tissues, we found two transcripts, one weakly expressed at approximately 4.0 kb and another more strongly expressed at approximately 2.6 kb, both hybridizing with two non-overlapping murine and rat eNOS probes. These results suggested the existence in the SCN of alternate transcripts for eNOS. We propose that two pathways could link light stimuli and NO release in the SCN: one involving N-methyl-D-aspartate (NMDA) receptors and nNOS in neurons; the other linking alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors and eNOS in astrocytes.
Caillol
M
M
Laboratoire de Biologie Cellulaire et Moléculaire, INRA, 78352 Jouy-en-Josas Cedex, France. caillol@jouy.inra.fr
Devinoy
E
E
Lacroix
M C
MC
Schirar
A
A
D017398:Alternative Splicing; D000818:Animals; D001253:Astrocytes / Q000201:enzymology; D015415:Biomarkers; D015153:Blotting, Western; D002940:Circadian Rhythm / Q000502:physiology; D006224:Cricetinae; D004790:Enzyme Induction / Q000528:radiation effects; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D017403:In Situ Hybridization; D008027:Light; D008297:Male; D008647:Mesocricetus / Q000033:anatomy & histology / Q000378:metabolism*; D051379:Mice; D009252:NADPH Dehydrogenase / Q000032:analysis; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009474:Neurons / Q000201:enzymology*; D009569:Nitric Oxide / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000032:analysis*; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats / Q000033:anatomy & histology / Q000378:metabolism*; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000502:physiology; D016194:Receptors, N-Methyl-D-Aspartate / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D013493:Suprachiasmatic Nucleus / Q000201:enzymology* / Q000528:radiation effects
D015415:Biomarkers; D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D018091:Receptors, AMPA; D016194:Receptors, N-Methyl-D-Aspartate; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496321:Nos1 protein, mouse; C496322:Nos1 protein, rat; C496315:Nos3 protein, mouse; C496316:Nos3 protein, rat; D009252:NADPH Dehydrogenase