Anatomical and temporal differences in the regulation of ZIF268 (NGFI-A) protein in the hamster and mouse suprachiasmatic nucleus.

Several immediate-early genes have been shown to be induced in the rodent circadian pacemaker, the suprachiasmatic nucleus, by retinal illumination at night. We compared spontaneous and light-evoked levels of the immediate-early gene protein ZIF268 (NGFI-A) in the Syrian hamster and C57BL/6J mouse suprachiasmatic nucleus. Exposure of both hamsters and mice to light pulses early and late in the subjective night caused increased ZIF268 immunoreactivity in the region of the suprachiasmatic nucleus that receives retinal innervation. In contrast to hamsters, mice also showed spontaneous increases in ZIF268 at both subjective night phases at the lateral edges of the suprachiasmatic nucleus. Light also evoked a significant increase in ZIF268 levels during the subjective day in the lateral suprachiasmatic nucleus, with few labeled cells in the ventral and dorsal suprachiasmatic nucleus. These results demonstrate a novel circadian pattern and regional differentiation of ZIF268 immunoreactivity in the suprachiasmatic nucleus of mice that differ from those in other rodents. There are pronounced species differences in both spontaneous and light-evoked expression of ZIF268 immunoreactivity.

Phase-shifting effects of pituitary adenylate cyclase activating polypeptide on hamster wheel-running rhythms.

Pituitary adenylate cyclase activating polypeptide (PACAP38) is a putative neurochemical of the main retinal input to the mammalian circadian pacemaker housed in the suprachiasmatic nucleus (SCN). We assessed the phase-resetting effects of microinjection of PACAP38 into the SCN region on hamster wheel-running rhythms. When administered during the middle of the subjective day, PACAP38 evoked large but transient phase advances ( approximately 60 min), that were followed by small, steady-state phase delays. During the early subjective night, PACAP38 elicited small to moderate phase delays without any detectable concentration-dependence. Late in the subjective night, PACAP38 had no significant effects. Saline microinjection had no effect at any phase tested. These findings show that PACAP38 has small to moderate effects on the phase of the hamster SCN circadian pacemaker, including significant phase delays early in the subjective night.

Daily rhythm of spontaneous immediate-early gene expression in the rat suprachiasmatic nucleus.

Nocturnal light induces the expression of various immediate-early genes (IEGs) in the suprachiasmatic nucleus (SCN), the primary pacemaker of the circadian system of mammals, and causes phase shifts of behavioral rhythms. In the hamster SCN, some IEGs show both sensitivity to light induction at night and a daily peak of spontaneous expression near dawn in different regions of the nucleus. To investigate whether both patterns of IEG expression are observed in the rat SCN, the authors studied the expression of NGFI-A, junB, c-fos, and fosB near the time of subjective dawn in rats entrained to a light-dark 12:12 cycle and then maintained in constant total darkness for approximately 48 h. They found that there were two independent rhythms of expression for junB and c-fos mRNAs in the SCN: (1) a rhythm of photic sensitivity expressed throughout the night and (2) a spontaneous rhythm of expression triggered around dawn and persisting for at least 2 h into the day. By contrast, fosB and NGFI-A transcripts were expressed only after light exposure at night and did not exhibit significant levels of spontaneous expression in the absence of photic input. These observations demonstrate that the circadian clock gates expression of two independent rhythms related to IEG expression in the rat SCN. The rhythm of sensitivity to nocturnal light exposure is expressed more strongly in the ventral SCN and may be related to photic entrainment. The second rhythm is triggered spontaneously in darkness around subjective dawn and is expressed in more dorsal parts of the SCN.

Circadian and photic regulation of immediate-early gene expression in the hamster suprachiasmatic nucleus.

The hypothalamic suprachiasmatic nucleus is the site of an endogenous circadian clock synchronized by daily light-dark cycles. At some daily phases, light exposure both shifts the clock and alters the expression of several immediate-early genes in cells of the suprachiasmatic nucleus. We have studied both spontaneous circadian and light-induced expression of several immediate-early gene messenger RNAs and proteins in hamsters in constant darkness or in response to brief light exposure. There was no detectable spontaneous expression of NGFI-A messenger RNA in suprachiasmatic nucleus cells at any circadian phase, but light pulses induced its expression selectively during the subjective night, with highest levels of expression 6 h into the night. We also found that there are two independent rhythms of expression of junB messenger RNA and JunB protein, as well as c-fos messenger RNA and c-Fos protein, in the suprachiasmatic nucleus of hamsters: a rhythm of photic sensitivity expressed throughout the night and a spontaneous rhythm of expression triggered around dawn. Induction of NGFI-A messenger RNA and c-fos messenger RNA and c-Fos protein in response to a light pulse were found throughout the suprachiasmatic nucleus, with the highest levels of expression in the ventrolateral subdivision; however, the spontaneous expression of JunB and c-Fos proteins was confined mainly to the dorsomedial suprachiasmatic nucleus. The temporal and anatomical differences in the expression of these immediate-early genes in the mammalian suprachiasmatic nucleus suggest that their protein products may be involved in different signaling mechanisms mediating either photic entrainment or endogenous oscillations within distinct subpopulations of suprachiasmatic nucleus cells.

Differential effects of glutamatergic blockade on circadian and photic regulation of gene expression in the hamster suprachiasmatic nucleus.

Nocturnal light exposure induces immediate-early gene (IEG) expression in the hypothalamic suprachiasmatic nucleus (SCN) and causes phase shifts of activity rhythms in mammals. Some IEGs also show a circadian rhythm of expression in the SCN. While excitatory amino acids (EAAs) are known to be involved in mediating photic regulation of entrainment and gene expression, their involvement in spontaneous rhythms of gene expression has not been studied. We assessed the role of NMDA receptors in the expression of NGFI-A, junB and fosB mRNAs induced by light pulses of different intensities late in the night (Zeitgeber Time [ZT] 18). We also examined the spontaneous expression of junB mRNA near subjective dawn (ZT 0). Both dim (5 lx) and bright (100 lx) light pulses induced similar levels of expression of NGFI-A and junB in the SCN late in the night. fosB mRNA was strongly induced by bright light but was less sensitive to dim light. At ZT 18, dizocilpine (MK-801) (3 mg/kg, i.p. ), a non-competitive NMDA receptor antagonist, almost completely blocked light-evoked expression of IEG mRNAs in the ventral SCN but not in the dorsolateral region at a mid-caudal level using either light intensity. At ZT 0, MK-801 strongly reduced light-evoked expression of junB mRNA in both SCN subdivisions, but inhibited spontaneous expression significantly only in the dorsal region. NMDA receptors appear to play an important role in mediating photic input regulating IEG expression only in the ventral SCN at night. At dawn, however, NMDA receptors are involved in mediating photic effects in both parts of the SCN, as well as being involved in spontaneous activation of junB expression selectively in the dorsal SCN. These findings support the idea that the effects in the dorsolateral SCN of nocturnal light exposure are mediated by different mechanisms than those in other portions of the nucleus.

Melatonin does not influence the expression of c-fos in the suprachiasmatic nucleus of rats and hamsters.

We have assessed whether melatonin can induce c-fos expression at various circadian phases, and whether melatonin can inhibit photically induced c-fos expression in the suprachiasmatic nucleus (SCN) in both rats and Syrian hamsters. Subcutaneous administration of melatonin at a dose of 100 microg/kg neither induced expression of Fos, the protein product of the c-fos proto-oncogene, nor inhibited the expression of Fos-like immunoreactivity (Fos-lir) induced by a light pulse in the SCN of rats and hamsters. In situ hybridization studies also demonstrated the absence of induction by acute melatonin treatments of c-fos mRNA in the SCN. Taken together, these results demonstrate that melatonin effects on SCN cells involve signal transduction pathways that do not include regulation of c-fos gene expression.

Spontaneous and light-evoked expression of JunB-like protein in the hamster suprachiasmatic nucleus near subjective dawn.

Some cells in the hamster suprachiasmatic nucleus (SCN) show a circadian rhythm of expression of junB mRNA in constant darkness, while others show junB mRNA only in response to light at night. We found that both the light-induced and spontaneous expressions of junB mRNA are translated into protein in SCN cells. In constant darkness, JunB-like immunoreactivity (lir) appears spontaneously in cells in the dorsal SCN around subjective dawn and persists for at least 4 h into the subjective day. During the subjective night, there is no spontaneous expression, but a light pulse can induce JunB-lir in cells throughout the SCN, and especially in the ventrolateral portion. As a component of AP-1 proteins, JunB may play a role both in mediating circadian responses to photic stimuli and in spontaneous oscillation of elements of the SCN circadian pacemaker.

Two distinct retinal projections to the hamster suprachiasmatic nucleus.

Electrical stimulation of the intergeniculate leaflet (IGL) region in hamsters can induce expression of Fos-like immunoreactivity (lir) in a restricted portion of the dorsolateral suprachiasmatic nucleus (SCN). The authors investigated whether the mechanisms by which stimulation affects SCN cells involves orthodromic activation of IGL cells projecting to the SCN or antidromic activation of retinal ganglion cells that send bifurcating axonal projections to both the IGL and the SCN. Bilateral optic enucleation strongly reduced induction of Fos-lir in SCN cells in response to electrical stimulation of the IGL region, implicating antidromic activation of retinal ganglion cells as the mechanism. This result implies that a class of retinal ganglion cells that project to the IGL also project selectively to the dorsolateral SCN. Earlier pharmacological studies suggest that this anatomically distinct retinal projection to the SCN is also neurochemically different from that innervating the rest of the nucleus.