0000000000243864

AUTHOR

Elisabeth Layes

Nitric oxide is formed in a subpopulation of rat pineal cells and acts as an intercellular messenger.

In the rat pineal, formation of the second messenger cyclic GMP (cGMP) is under adrenergic control. Two important sequential steps mediate adrenergic signal transduction by cGMP, receptor-stimulated nitric oxide (NO) formation by the enzyme NO synthase I (NOS I), and NO-induced cGMP formation by the cytosolic enzyme guanylyl cyclase. With regard to the first step in cGMP transduction (i.e. NO formation) we found, by means of NOS I immunostaining and NADPH-diaphorase staining, that the presence of NOS I was restricted to a subpopulation of pineal cells, generally surrounded by NOS I-negative cells. Considering the fact that NO is able to permeate the cell membrane, the question arises whethe…

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Nitric oxide synthase in the hypothalamic suprachiasmatic nucleus of rat: evidence from histochemistry, immunohistochemistry and Western blot; and colocalization with VIP

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 mo…

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Down-regulation of the nocturnally elevated guanylyl cyclase activity in the rat pineal gland

Abstract Previous studies have shown that in the rat pineal, the cytosolic and the particulate forms of guanylyl cyclase (GC) activity undergo a biphasic 24-h rhythm with two prominent peaks, one in the middle of the light phase and the other in the middle of the dark phase. In this study we investigated whether the well established photo-neural adrenergic regulatory processes identified for pineal melatonin synthesis also apply to the nocturnal elevation of GC activity. A 10-min light pulse given in the middle of the dark phase decreases the cytosolic and the particulate forms of GC. Administration of the β-receptor blocker propranolol did not depress the nocturnally elevated GC activity. …

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Adrenergic Stimulation of Cyclic GMP Formation Requires NO-Dependent Activation of Cytosolic Guanylate Cyclase in Rat Pinealocytes

Cyclic GMP (cGMP) formation in rat pinealocytes is regulated through a synergistic dual receptor mechanism involving beta- and alpha 1-adrenergic receptors. The effects of NG-monomethyl-L-arginine (NMMA), which inhibits nitric oxide (NO) synthase and NO-mediated activation of cytosolic guanylate cyclase, and methylene blue (MB), which inhibits cytosolic guanylate cyclase, were investigated in an attempt to understand the role of NO in adrenergic cGMP formation. Both NMMA and MB inhibited beta-adrenergic stimulation of cGMP formation as well as alpha 1-adrenergic potentiation of beta-adrenergic stimulation of cGMP formation, whereas they had no effect in unstimulated pinealocytes. The inhibi…

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Expression of Nitric Oxide Synthase I Determines the Strength of Adrenergic Cyclic GMP Response in the Rat Pineal Gland

In the rat pineal gland the adrenergic cGMP response is not static but changes during postnatal development and depends on environmental lighting conditions. Since the adrenergic cGMP transduction is mediated by stimulation of the enzyme nitric oxide synthase I (NOS I), we investigated whether changes in the strength of adrenergic cGMP response coincide with the expression of NOS I. Using immunoblot analysis, we found that the increase in adrenergic cGMP transduction in the second postnatal week coincides with enhanced NOS I expression. Similarly, we observed that changes in the intensity of adrenergic cGMP transduction under different environmental lighting conditions correspond to NOS I e…

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