Nitric oxide synthase in the hypothalamic suprachiasmatic nucleus of rat: evidence from histochemistry, immunohistochemistry and Western blot; and colocalization with VIP |
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Authors: | Stefan Reuss,Karin Decker,Lukas Rö βeler,Elisabeth Layes,Andreas Schollmayer,Rainer Spessert |
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Affiliation: | Department of Anatomy, School of Medicine, Johannes Gutenberg-University, D-55099 Mainz, Germany |
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Abstract: | Nitric oxide (NO) is a neuroactive substance of high potency. Physiological results revealed the involvement of NO in circadian regulation of rats. Since neuronal structures containing NO-synthase (NOS) were previously not found in the circadian oscillator, the hypothalamic suprachiasmatic nucleus (SCN), in this species but are present in the hamster, we investigated the distribution of NO-producing structures in the rat SCN by Western blot analysis, immunohistochemistry of NOS, and by histochemistry (NADPH-diaphorase (NADPH-d) activity of NOS). Western blot analysis of SCN homogenates from rat (and, for comparison, hamster) showed a NOS-like immunoreactive (-LI) protein band of apparent molecular mass of 150 kDa, consistent with the neuronal NOS molecule. In the rat SCN, perikarya exhibiting NADPH-d staining or NOS-LI with a complete overlapping of both were found. Double-immunofluorescence experiments revealed that NOS cells are a subgroup of the neuronal SCN population that is characterized by immunoreactivity to vasoactive intestinal polypeptide. These data provide evidence for the existence of neuronal nitric oxide synthase in the rat SCN and may explain the involvement of NO in the mediation of photic information. |
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Keywords: | Hypothalamus Neuronal nitric oxide synthase Hamster NADPH-diaphorase Western blot Circadian system Vasoactive intestinal polypeptide |
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