Organotypic cultures of the rat hypothalamus are very useful models for the long-term study of parvocellular vasopressin (VP) neurons in the paraventricular (PVN) and suprachiasmatic (SCN) nuclei. However, they do not preserve significant numbers of VP magnocellular neurons (VP-MCNs) in either the PVN or the supraoptic nucleus (SON). Vutskits et al. [(1998) Neuroscience 87:571-582] reported that ciliary neurotrophic factor (CNTF) was a selective survival factor for rat VP-MCNs in organotypic cultures of the rat hypothalamic paraventricular nucleus (PVN). We examined the effects of CNTF on the survival of these neurons in rat and mouse SONs. CNTF (10 ng/ml) in the culture media increased the survival of VP-MCNs by 6-fold and OT-MCNs by 3-fold. In the mouse, both OT- and VP-MCNs survive very well in organotypic cultures under standard culture conditions and the addition of CNTF had no further effect. Consistent with these results, in situ hybridization showed substantially higher levels of VP- and OT-mRNA in rat PVNs and SONs in the presence of CNTF, but produced no changes in these nuclei in the mouse. The optimum period for the survival effect of CNTF on MCNs in the rat hypothalamic cultures was in the first 7-10 days of culture and this effect is maintained for at least 5 additional days if CNTF is then removed from the medium. Therefore, using CNTF in the culture media can provide an opportunity for long-term studies of rat VP- and OT-MCNs in SONs in organotypic cultures.
Copyright 2002 Wiley-Liss, Inc.