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Journal of Biological Rhythms
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Light-Dependent Induction of cFos during Subjective Day and Night in PACAP-Containing Ganglion Cells of the Retinohypothalamic Tract

Jens Hannibal

Department of Clinical Biochemistry, Bispebjerg Hospital, University of Copenhagen, Denmark; Bispebjerg Bakke 23, DK-2400 Copenhagen NV, Denmark; j.hannibal{at}inet.uni2.dk

Niels Vrang

Department of Medical Anatomy, The Panum Institute, University of Copenhagen, Denmark

J. Patrick Card

Departments of Neuroscience and Psychiatry, University of Pittsburgh, Pittsburgh, PA 15260, USA

Jan Fahrenkrug

Department of Clinical Biochemistry, Bispebjerg Hospital, University of Copenhagen, Denmark

Environmental light stimulation via the retinohypothalamic tract (RHT) is necessary for stable entrainment of circadian rhythms generated in the suprachiasmatic nucleus (SCN). In the current report, the authors characterized the functional activity and phenotype of retinal ganglion cells that give rise to the RHT of the rat. Retinal ganglion cells that give rise to the RHT were identified by transsynaptic passage of an attenuated alpha herpesvirus known to have selective affinity for this pathway. Dual labeling immunocytochemistry demonstrated co-localization of viral antigen and pituitary adenylate cyclase activating polypeptide (PACAP) in retinal ganglion cells. This was confirmed using the anterograde tracer cholera toxin subunit B (ChB). In normal and retinally degenerated monosodium glutamate (MSG)–treated rats, ChB co-localized with PACAP in axons of the retinorecipient zone of the SCN. Light-induced Fos-immunoreactivity (Fos-IR) was apparent in all PACAP-containing retinal ganglion cells and a population of non-PACAP-containing retinal ganglion cells at dawn of normal and MSG-treated animals. Within the next 3 h, Fos disappeared in all non-PACAP-immunoreactive cells but persisted in all PACAP- containing retinal ganglion cells until dusk. When animals were exposed to constant light, Fos-IR was sustained only in the PACAP-immunoreactive (PACAP-IR) retinal ganglion cells. Darkness eliminated Fos-IR in all PACAP-IR retinal ganglion cells, demonstrating that the induction of Fos gene expression was light dependent. When animals were maintained in constant darkness and exposed to light pulses at ZT 14, ZT 19, or ZT 6, Fos-IR was induced in PACAP-IR retinal ganglion cells in a pattern similar to that seen at dawn. Collectively, these data indicate that PACAPis present in ganglion cells that give rise to the RHT and suggest a role for this peptide in the light entrainment of the clock

Key Words: PRV Bartha virus • monosodium glutamate • co-localization • suprachiasmatic nucleus • circadian rhythm • rat

Journal of Biological Rhythms, Vol. 16, No. 5, 457-470 (2001)
DOI: 10.1177/074873001129002132


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