A confocal approach to the morphofunctional characterization of the transient tyrosine hydroxylase system in the rat suprachiasmatic nucleus

Abstract

The suprachiasmatic nucleus (SCN) of the neonatal rat is transiently innervated by tyrosine hydroxylase (TH) fibers of unknown origin and whose catecholaminergic nature is rather doubtful. In order to characterize this system morphofunctionally, immunocytochemical double labelling and confocal laser scanning microscopy analysis were employed on cryostat brain sections of 10-day-old rats. Simultaneous stainings for neuropeptide Y (NPY) and tyrosine hydroxylase (TH) immunoreactivity showed that they are not colocalized, neither in the SCN fibers nor in the intergeniculate leaflet (IGL) neurons, site of origin of the NPY projection to the SCN. Therefore, the possibility that SCN transient TH fiber system originates from the IGL could be excluded. Double labelling for TH and aromatic l-aminoacid decarboxylase (AADC) demonstrated that transient SCN TH immunoreactive (IR) fibers are AADC negative, thus supporting the hypothesis of their non-catecholaminergic nature. Moreover two new group of cells which are TH positive and AADC negative were found: one in the SCN and the other in the periventricular hypothalamic nucleus (PHN). The presence of somatostatin (SRIF) and TH in PHN neurons and SCN fibers suggested their possible colocalization, but double immunolabellings gave negative results. Simultaneous immunocytochemical staining for vasoactive intestinal polypeptide (VIP) and TH showed that TH fibers may interact with ventrolateral SCN VIP neurons. This result suggests a possible involvement of TH fibers in regulating VIP cells activity in the entrainment of circadian rhythms.