Daily variation of muscarinic receptors in visual cortex but not suprachiasmatic nucleus of Syrian hamsters.

Intraventricular administration of carbachol can induce phase shifts in wheel-running activity in rodents, which depend on circadian phase and are mediated via muscarinic cholinergic receptors in Syrian hamsters. We studied the circadian variation in binding of [3H]-N-methylscopolamine ([3H]NMS), a hydrophilic muscarinic receptor antagonist, in micropunches obtained from the anterior hypothalamus and occipital cortex of Syrian hamsters housed in a 14:10 light:dark cycle. Binding sites were characterized on cells contained within 1 mm punches (obtained from slices 300 microm thick), using a method to selectively detect cell surface (functional) receptors. Atropine sulphate was used to determine nonspecific binding. Cortex showed a significant daily rhythm in [3H]NMS binding with a peak occurring late in the light phase and a trough at lights on, while the hypothalamus showed no detectable rhythm. Following suprachiasmatic nucleus (SCN) ablation or maintenance in constant darkness, the rhythm in the cortex was abolished. These findings suggest that photic information conveyed via the SCN is responsible for the receptor binding rhythm in the cortex. Autoradiographic studies ([3H]NMS; 2 nM, 3 weeks exposure) clearly revealed both M1 and M2 subtypes of muscarinic receptors in the region of the SCN and the visual cortex.

Distribution of pituitary adenylate cyclase activating polypeptide (PACAP) immunoreactivity in the hypothalamus and extended amygdala of the rat.

Pituitary adenylate cyclase activating polypeptide (PACAP) is found in two forms of 27 and 38 amino acids (PACAP-27 and PACAP-38 respectively) in the mammalian central nervous system. Using antibodies to these two forms of PACAP, we examined the distribution of PACAP immunoreactivity in the rat hypothalamus and a number of extrahypothalamic areas. The patterns of immunostaining for PACAP-27 and PACAP-38 were similar: prominent terminal labelling was present in the retrochiasmatic area, median eminence, and posterior periventricular nucleus of the hypothalamus as well as the bed nucleus of the stria terminalis and amygdaloid complex. After colchicine treatment, immunopositive cell bodies were found in the preoptic region of the periventricular zone of the hypothalamus, the suprachiasmatic and paraventricular hypothalamic nuclei, neural structures adjacent to the median eminence (including the retrochiasmatic area, arcuate nucleus, ventromedial hypothalamus, and tuber cinereum), and the lateral mammillary and supramammillary nuclei. In all these areas, immunolabelling appeared specific since it was abolished by preabsorption of primary antisera with the appropriate PACAP peptide. However, the number of immunopositive cells in the suprachiasmatic nucleus was also reduced by preabsorption of PACAP-27/38 antisera with vasoactive intestinal polypeptide, suggesting that a subpopulation of cells in the suprachiasmatic nucleus express a peptide which has significant sequence homology with both PACAP-27/38 and vasoactive intestinal polypeptide. The distribution of PACAP immunoreactivity throughout the hypothalamus, bed nucleus of the stria terminalis, and amygdala suggests the involvement of PACAP in a number of processes including limbic, autonomic, and neuroendocrine functions as well as regulation of the circadian pacemaker.

Hamster circadian rhythms are phase-shifted by electrical stimulation of the geniculo-hypothalamic tract.

The suprachiasmatic nuclei (SCN) contain the major pacemaker for mammalian circadian rhythms. The SCN receive photic input both directly, via the retinohypothalamic tract (RHT), and indirectly, via the geniculohypothalamic tract (GHT), which originates in cells in the intergeniculate leaflet (IGL) and anterior portions of the ventral lateral geniculate nucleus (vLGN). We tested whether electrical stimulation of the GHT would induce phase shifts in wheel-running activity rhythms of Syrian hamsters housed in continuous darkness or continuous illumination. In both lighting conditions, electrical stimulation of the GHT induced mainly phase advances when given during the late subjective day and small phase delays when given during the late subjective night and early subjective day. Stimulation in the thalamus outside the GHT failed to produce similar phase shifts. Repeated daily stimulation had only a weak entraining effect on the activity rhythm. Activation of GHT neurons appears to influence the pacemaker for activity rhythms in a phase-dependent manner.

Photic responses of geniculo-hypothalamic tract neurons in the Syrian hamster.

The putative neural pacemaker controlling circadian rhythms in mammals is contained in the suprachiasmatic nuclei of the hypothalamus. These nuclei receive a projection, the geniculo-hypothalamic tract (GHT), from neurons in the intergeniculate leaflet (IGL) and portions of the ventral lateral geniculate nucleus (vLGN) of the thalamus. We examined the responses of putative GHT neurons to diffuse illumination using extracellular electrophysiological recordings. The great majority of IGL neurons showed sustained ON responses to diffuse retinal illumination; vLGN neurons showed more variation in their responses. Discharge rates of sustained ON neurons increased monotonically as light intensity was increased and saturated over 2-3 log units of intensity changes. Many IGL neurons had binocular input, and input from the ipsilateral eye was often inhibitory. These results indicate that GHT neurons may provide information about ambient light intensity to the suprachiasmatic nuclei.

Ablation of the geniculo-hypothalamic tract alters circadian activity rhythms of hamsters housed under constant light.

Retino-recipient cells in the hamster lateral geniculate nucleus project to the suprachiasmatic nucleus via the geniculo-hypothalamic tract (GHT). GHT-ablation alters phase advance shifts to light pulses in a hamster's late subjective night. In this study, the effects of GHT-ablation on wheel-running rhythms of hamsters housed under continuous illumination (LL) were assessed. In the first experiment, hamsters received GHT-ablation or sham surgery while under a light:dark schedule and were subsequently exposed to 250 days of LL. GHT-ablated hamsters showed rhythms with shorter periods and were less likely to show split activity rhythms than sham-operated or partial-lesion controls. In the second experiment, hamsters were housed under LL until rhythms split into two components; hamsters then received either GHT-ablation or sham surgery. Four of seven GHT-ablated hamsters showed re-fusion of their activity pattern into one component, while none of the eight sham-operated animals showed such re-fusion. The results of these two experiments indicate that GHT-ablation alters the responsiveness of the activity rhythm pacemaker to LL exposure.

Suprachiasmatic stimulation phase shifts rodent circadian rhythms.

The integrity of the suprachiasmatic nuclei (SCN) of the hypothalamus is essential to the expression of normal circadian rhythms in rodents. Electrical stimulation of the SCN caused phase shifts and period changes in the freerunning feeding rhythms of rats and activity rhythms of hamsters. The phase response curve for SCN stimulation appears to parallel that for light pulses. These findings strengthen the hypothesis derived from lesion studies that the SCN are the dominant light-entrained oscillators in the rodent circadian system.

Neural mechanisms for entrainment and generation of mammalian circadian rhythms.

The identification of a direct retinohypothalamic tract (RHT) terminating in the supra-chiasmatic nuclei (SCN) has focused attention on the role of these structures in the entrainment and generation of circadian rhythms in mammals. Light effects on circadian rhythms are mediated by both the RHT and portions of the classical visual system. The complex interactions of these systems are reflected both in their direct anatomical connections and in the functional changes in entrainment produced by interruption of either set of projections. Destruction of the RHT/SCN eliminated both normal entrainment and normal free-running circadian rhythms. No circadian rhythms has survived SCN ablation in rodents, but a variety of non-circadian cycles can be generated by lesioned animals. The complex behavioral patterns produced by SCN-lesioned hamsters suggest that circadian oscillators continue to function in these animals, but that their activity is no longer integrated into a single circadian framework. The available evidence indicates that the mammalian pacemaking system comprises a set of independent oscillators normally regulated by the SCN and by light information that is transmitted via several retinofugal pathways.