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Department of Medical Anatomy, University of Copenhagen (N.V., P.J.L., M.T.-C.), Blegdamsvej 3, 2200 Copenhagen; and Department of Histology, Novo Nordisk A/S (P.K., M.-T.C.), 2880 Bagsvaerd, Denmark
Address all correspondence and requests for reprints to: Niels Vrang, M.D., Department of Medical Anatomy, B, The Panum Institute, University of Copenhagen, Blegdamsvej 3, DK-2200 Copenhagen N, Denmark. E-mail: n.vrang{at}mai.ku.dk
We have recently shown that intracerebroventricular (icv) administration of the hypothalamic neuropeptide cocaine-amphetamine-regulated transcript (CART) inhibits food intake and induces the expression of c-fos in several nuclei involved in the regulation of food intake. A high number of CART-induced c-Fos-positive nuclei in the paraventricular nucleus of the hypothalamus prompted us to examine the effect of icv recombinant CART-(42–89) on activation of CRH-, oxytocin-, and vasopressin-synthesizing neuroendocrine cells in the paraventricular nucleus (PVN). In addition, plasma levels of glucose were examined after central administration of CART-(42–89). Seventy-six male Wistar rats were fitted with icv cannulas and singly housed under 12-h light, 12-h dark conditions. One week postsurgery the animals were injected icv in the morning with 0.5 µg recombinant CART-(42–89) or saline. Trunk blood was collected by decapitation at 0 (baseline), 10, 20, 40, 60, 120, or 240 min. CART caused a strong increase in circulating corticosterone that was significantly different from saline at 20, 40, 60, and 120 min postinjection (P < 0.05). Furthermore, CART caused a transient rise in plasma oxytocin levels (P < 0.05 at 10 and 20 min postinjection), whereas plasma vasopressin levels were unaffected by icv CART. Animals injected icv with CART showed a rise in blood glucose levels 10 min postinjection (P < 0.05). To examine whether the stimulatory effect of icv CART on corticosterone and oxytocin secretion is caused by activation of paraventricular nucleus/supraoptic nucleus (PVN/SON) neuroendocrine neurons, we used c-Fos as a marker of neuronal activity. Animals injected with CART showed a strong increase in c-Fos-immunoreactive nuclei in the PVN. Double immunohistochemistry revealed that a high (89 ± 0.4%) number of CRH-immunoreactive neurons in the PVN contained c-Fos after CART icv. c-Fos expression was also observed in oxytocinergic cells (in both magnocellular and parvicellular PVN neurons as well as in the supraoptic nuclei) 120 min after CART administration, whereas none of the vasopressinergic neurons contained c-Fos. Triple immunofluorescence microscopy revealed that CART-immunoreactive fibers closely apposed c-Fos-positive CRH neurons, suggestive of a direct action of CART on PVN CRH neurons. In summary, icv CART activates central CRH neurons as well as both magnocellular (presumably neurohypophysial) and parvicellular (presumably descending) oxytocinergic neurons of the PVN. The effect of CART on CRH neurons most likely leads to corticosterone secretion from the adrenal gland, which may contribute to the inhibitory effects of CART on feeding behavior.
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