title: Alpha-2 adrenergic regulation of pedunculopontine nucleus neurons during development.
abstract: Rapid eye movement sleep decreases between 10 and 30 days postnatally in the rat. The pedunculopontine nucleus is known to modulate waking and rapid eye movement sleep, and pedunculopontine nucleus neurons are thought to be hyperpolarized by noradrenergic input from the locus coeruleus. The goal of the study was to investigate the possibility that a change in alpha-2 adrenergic inhibition of pedunculopontine nucleus cells during this period could explain at least part of the developmental decrease in rapid eye movement sleep. We, therefore, recorded intracellularly in 12-21 day rat brainstem slices maintained in oxygenated artificial cerebrospinal fluid. Putative cholinergic vs. non-cholinergic pedunculopontine nucleus neurons were identified using nicotinamide adenine dinucleotide phosphate diaphorase histochemistry and intracellular injection of neurobiotin (Texas Red immunocytochemistry). Pedunculopontine nucleus neurons also were identified by intrinsic membrane properties, type I (low threshold spike), type II (A) and type III (A+low threshold spike), as previously described. Clonidine (20 microM) hyperpolarized most cholinergic and non-cholinergic pedunculopontine nucleus cells. This hyperpolarization decreased significantly in amplitude (mean+/-S.E.) from -6.8+/-1.0 mV at 12-13 days, to -3.0+/-0.7 mV at 20-21 days. However, much of these early effects (12-15 days) were indirect such that direct effects (tested following sodium channel blockade with tetrodotoxin (0.3 microM)) resulted in hyperpolarization averaging -3.4+/-0.5 mV, similar to that evident at 16-21 days. Non-cholinergic cells were less hyperpolarized than cholinergic cells at 12-13 days (-1.6+/-0.3 mV), but equally hyperpolarized at 20-21 days (-3.3+/-1.3 mV). In those cells tested, hyperpolarization was blocked by yohimbine, an alpha-2 adrenergic receptor antagonist (1.5 microM). These results suggest that the alpha-2 adrenergic receptor on cholinergic pedunculopontine nucleus neurons activated by clonidine may play only a modest role, if any, in the developmental decrease in rapid eye movement sleep. Clonidine blocked or reduced the hyperpolarization-activated inward cation conductance, so that its effects on the firing rate of a specific population of pedunculopontine nucleus neurons could be significant. In conclusion, the alpha-2 adrenergic input to pedunculopontine nucleus neurons appears to consistently modulate the firing rate of cholinergic and non-cholinergic pedunculopontine nucleus neurons, with important effects on the regulation of sleep-wake states.
authors.affiliation: Center for Translational Neuroscience, Department of Neurobiology and Dev. Sci., College of Medicine, University of Arkansas for Medical Sciences, 4301 West Markham Street, Slot 847, Little Rock, AR 72205, USA.
authors.forename: K D
authors.initials: KD
authors.lastname: Bay
authors.affiliation: Center for Translational Neuroscience, Department of Neurobiology and Dev. Sci., College of Medicine, University of Arkansas for Medical Sciences, 4301 West Markham Street, Slot 847, Little Rock, AR 72205, USA.
authors.forename: K
authors.initials: K
authors.lastname: Mamiya
authors.affiliation: Center for Translational Neuroscience, Department of Neurobiology and Dev. Sci., College of Medicine, University of Arkansas for Medical Sciences, 4301 West Markham Street, Slot 847, Little Rock, AR 72205, USA.
authors.forename: C H
authors.initials: CH
authors.lastname: Good
authors.affiliation: Center for Translational Neuroscience, Department of Neurobiology and Dev. Sci., College of Medicine, University of Arkansas for Medical Sciences, 4301 West Markham Street, Slot 847, Little Rock, AR 72205, USA.
authors.forename: R D
authors.initials: RD
authors.lastname: Skinner
authors.affiliation: Center for Translational Neuroscience, Department of Neurobiology and Dev. Sci., College of Medicine, University of Arkansas for Medical Sciences, 4301 West Markham Street, Slot 847, Little Rock, AR 72205, USA. Electronic address: GarciaRillEdgar@uams.edu.
authors.forename: E
authors.initials: E
authors.lastname: Garcia-Rill
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000200:Action Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D000316:Adrenergic alpha-Agonists / Q000494:pharmacology*; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000704:Analysis of Variance; D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D001710:Biotin / Q000031:analogs & derivatives / Q000378:metabolism; D003000:Clonidine / Q000494:pharmacology*; D004558:Electric Stimulation / Q000379:methods; D005260:Female; D066298:In Vitro Techniques; D008297:Male; D009249:NADP / Q000378:metabolism; D009474:Neurons / Q000145:classification / Q000187:drug effects* / Q000502:physiology / Q000528:radiation effects; D045042:Pedunculopontine Tegmental Nucleus / Q000166:cytology* / Q000254:growth & development; D011247:Pregnancy; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013779:Tetrodotoxin / Q000494:pharmacology; D015016:Yohimbine / Q000494:pharmacology
chemical_list: D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D000779:Anesthetics, Local; D011743:Pyrimidines; C070614:neurobiotin; C082246:ICI D2788; D015016:Yohimbine; D013779:Tetrodotoxin; D009249:NADP; D001710:Biotin; D003000:Clonidine; D000109:Acetylcholine