Merkel Cells as Putative Regulatory Cells in Skin Disorders: An In Vitro Study

Merkel cells (MCs) are involved in mechanoreception, but several lines of evidence suggest that they may also participate in skin disorders through the release of neuropeptides and hormones. In addition, MC hyperplasias have been reported in inflammatory skin diseases. However, neither proliferation nor reactions to the epidermal environment have been demonstrated. We established a culture model enriched in swine MCs to analyze their proliferative capability and to discover MC survival factors and modulators of MC neuroendocrine properties. In culture, MCs reacted to bFGF by extending outgrowths. Conversely, neurotrophins failed to induce cell spreading, suggesting that they do not act as a growth factor for MCs. For the first time, we provide evidence of proliferation in culture through Ki-67 immunoreactivity. We also found that MCs reacted to histamine or activation of the proton gated/osmoreceptor TRPV4 by releasing vasoactive intestinal peptide (VIP). Since VIP is involved in many pathophysiological processes, its release suggests a putative regulatory role for MCs in skin disorders. Moreover, in contrast to mechanotransduction, neuropeptide exocytosis was Ca2+-independent, as inhibition of Ca2+ channels or culture in the absence of Ca2+ failed to decrease the amount of VIP released. We conclude that neuropeptide release and neurotransmitter exocytosis may be two distinct pathways that are differentially regulated

Evidence of Collateralization of Vagal Efferents Innervating Subdiaphragmatic Segments of the Gastrointestinal Tract in the Rat Using the Double Labelling Fluorescent Dyes Technique

Collateralization of the abdominal vagal trunks was investigated in the rat using double labelling fluorescence dye technique. A total of 20 adult male and female rats were used for the study. The anterior and posterior walls of the corpus and fundus of the stomach were injected with 0.1 microlitre(µl) of 5% aqueous solution of diamidino yellow (Dy) in eight rats. The same quantity and percentage of fast blue (Fb) was injected into the walls of the duodenum and proximal jejunum in the same eight rats that were injected with Dy. The anterior and posterior walls of the stomach of four rats were injected with 0.1µl of 5% Dy only while four other rats had injections of 0.1µl of Fb only into the duodenum and proximal jejunum. Two control rats had stomach injections with normal saline, and two rats had saline injections into the intestine. Each rat was perfused with a preservative 14 days after injection and serial sections taken for examination with a fluorescence microscope. The result of the experiment revealed that in the eight rats injected simultaneously with Dy and Fb, some neurons of the dorsal motor nucleus of the vagus nerve (DMX) were labelled with Dy only, some with Fb only and some were doubly labelled with Dy and Fb. No double-labelled neurons were seen in the rat injected with one dye only and no labellings at all were seen in the controls. The pattern of labelling obtained in the study is suggestive of collateralization of axons of the abdominal vagal trunks