Role of membrane conductances and protein synthesis in subjective day phase advances of the hamster circadian clock by neuropeptide Y.
Neurons of the mammalian circadian pacemaker in the hypothalamic suprachiasmatic nuclei exhibit a rhythm in firing rate that can be reset by neuropeptide Y. We recorded the effects of neuropeptide Y on Na+ and K+ conductances of hamster suprachiasmatic nuclei neurons using whole-cell, perforated-patch and cell-attached patch-clamp recordings, both in dissociated and brain slice preparations. While neuropeptide Y had no effect on voltage-gated Na+ currents, neuropeptide Y activated a leak K+ current. Neuropeptide Y phase advances in the suprachiasmatic nuclei brain slice preparation were blocked by a number of K+ channel blockers (tetraethylammonium chloride, dendrotoxin-I, glybenclamide). However, a K+ ionophore, valinomycin, did not shift the rhythm. The inhibition by tetraethylammonium chloride did not persist in the presence of glutamatergic receptor blockers. We have previously shown that glutamate can oppose neuropeptide Y phase-shifting actions, suggesting that K+ channel inhibition acts by inducing glutamate release. Protein synthesis inhibitors had no effect on clock phase when applied during the subjective day, and had no influence on neuropeptide Y-induced phase shifts. On the other hand, glutamate's ability to inhibit neuropeptide Y shifts was abolished by protein synthesis inhibition. Thus, while neuropeptide Y phase shifts do not require protein synthesis, glutamate blocks neuropeptide Y shifts via increased gene expression during the subjective day, at a time when it does not reset the clock. These results indicate that neuropeptide Y phase shifts via a mechanism that does not involve changes in membrane conductance or protein synthesis.
Hall
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Department of Psychology, Smith College, Northampton, MA 01063, USA.
Earle-Cruikshanks
G
G
Harrington
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D015763:2-Amino-5-phosphonovalerate / Q000494:pharmacology; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D002940:Circadian Rhythm / Q000187:drug effects*; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D004553:Electric Conductivity; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018698:Glutamic Acid / Q000502:physiology; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008647:Mesocricetus; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D009474:Neurons / Q000737:chemistry / Q000502:physiology; D009478:Neuropeptide Y / Q000494:pharmacology*; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000502:physiology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism; D015222:Sodium Channels / Q000502:physiology; D013493:Suprachiasmatic Nucleus / Q000166:cytology; D019789:Tetraethylammonium / Q000494:pharmacology; D014634:Valinomycin / Q000494:pharmacology
D018691:Excitatory Amino Acid Antagonists; D007476:Ionophores; D009419:Nerve Tissue Proteins; D009478:Neuropeptide Y; D015221:Potassium Channels; D011500:Protein Synthesis Inhibitors; D015222:Sodium Channels; D014634:Valinomycin; D018698:Glutamic Acid; D019789:Tetraethylammonium; D000841:Anisomycin; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D015763:2-Amino-5-phosphonovalerate; D003513:Cycloheximide; D012964:Sodium; D011188:Potassium