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Volume 17, Number 7, Issue of April 1, 1997 pp. 2637-2644
Copyright ©1997 Society for Neuroscience

Pituitary Adenylate Cyclase-Activating Peptide (PACAP) in the Retinohypothalamic Tract: A Potential Daytime Regulator of the Biological Clock

Received Aug. 22, 1996; revised Jan. 17, 1997; accepted Jan. 23, 1997.

Jens Hannibal1, Jian M. Ding3, Dong Chen4, Jan Fahrenkrug1, Philip J. Larsen2, Martha U. Gillette3, 4, and Jens D. Mikkelsen2, 5

1 Department of Clinical Biochemistry, Bispebjerg Hospital, University of Copenhagen, DK-2400 Copenhagen NV, Denmark, 2 Institute of Medical Anatomy, University of Copenhagen, DK-2400 Copenhagen NV, Denmark, Departments of 3 Cell and Structural Biology and 4 Molecular and Integrative Physiology, University of Illinois, Urbana, Illinois, and 5 Department of Neurobiology, Research and Development, H. Lundbeck A/S, DK-2500 Valby Copenhagen, Denmark

The retinohypothalamic tract (RHT) relays photic information from the eyes to the suprachiasmatic nucleus (SCN). Activation of this pathway by light plays a role in adjusting circadian timing via a glutamatergic pathway at night. Here we report a new signaling pathway by which the RHT may regulate circadian timing in the daytime as well. We used dual immunocytochemistry for pituitary adenylate cyclase-activating peptide (PACAP) and the in vivo tracer cholera toxin subunit B and observed intense PACAP-immunoreactivity (PACAP-IR) in retinal afferents in the rat SCN as well as in the intergeniculate leaflet (IGL) of the thalamus. This PACAP-IR in the SCN as well as in the IGL was nearly lost after bilateral eye enucleation. PACAP afferents originated from small ganglion cells distributed throughout the retina. The phase of circadian rhythm measured as SCN neuronal activity in vitro was significantly advanced (3.5 ± 0.4 hr) by application of 1 × 10-6 M PACAP-38 during the subjective day [circadian time (CT)-6] but not at night (CT14 and CT19). The phase-shifting effect is channeled to the clock via a PACAP-R1 receptor, because mRNA from this receptor was demonstrated in the ventral SCN by in situ hybridization. Furthermore, vasoactive intestinal peptide was nearly 1000-fold less potent in stimulating a phase advance at CT6. The signaling mechanism was through a cAMP-dependent pathway, which could be blocked by a specific cAMP antagonist, Rp-cAMPS. Thus, in addition to its role in nocturnal regulation by glutamatergic neurotransmission, the RHT may adjust the biological clock by a PACAP/cAMP-dependent mechanism during the daytime.

Key words: suprachiasmatic nucleus; circadian; phase shifting; brain slice; PACAP receptor; cAMP; rat; intergeniculate leaflet; ganglion cells




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