PMID- 10048470 OWN - NLM STAT- MEDLINE DCOM- 19990421 LR - 20190921 IS - 0953-8194 (Print) IS - 0953-8194 (Linking) VI - 11 IP - 2 DP - 1999 Feb TI - Effects of osmotic pressure and brain-derived neurotrophic factor on the survival of postnatal hypothalamic oxytocinergic and vasopressinergic neurons in dissociated cell culture. PG - 145-52 AB - Neurons from hypothalamic paraventricular nuclei (PVN) and supraoptic nuclei (SON) from postnatal day 6-8 rats were enzymatically dissociated and separately maintained in monolayer cultures for 14 days. The osmotic pressure of the culture medium, based on Neurobasal medium (Life Technologies), was varied (255, 300 and 330 mOsm/l) by adjustment using mannitol. The survival of oxytocin (OT), vasopressin (VP) and oxytocin-vasopressin (OT/VP) coexpressing neurons were studied under these varied conditions, and the identification of the cell phenotypes in the cultures was carried out by using double-label immunofluorescence. Under control osmolar conditions (300 mOsm/l) equivalent numbers of OT and VP neurons were found in the SON (P = 0.8398) and PVN (P = 0.4721) cultures. The OT neurons' survival did not change in 255 or 330 mOsm media in the SON cultures, but the VP neurons in the SON cultures were significantly increased in 255 mOsm/l medium as compared to control (300 mOsm/l) medium (P = 0.0088). No significant changes were found in VP neuron survival in SON cultures between the 300-330 mOsm/l media (P = 0.2372). Similar data were obtained for the VP neurons in PVN-derived cultures, but the OT neurons in these cultures survived significantly better at 300 mOs/l than at 255 mOsm/l (P<0.0001), but were not significantly different at 330 mOsm/l (P = 0.1208). In general, the VP neurons were more vulnerable than OT neurons to increases of culture medium osmolarity with respect to their survival. The number of OT/VP coexpressing neurons was greater in SON-derived cell cultures as compared to PVN-derived cell cultures, and their numbers were higher in the lower osmolarity media. The effects of adding brain-derived neurotrophic factor (BDNF) to the culture medium on survival were determined. BDNF significantly increased the numbers of all three types of neurons in both PVN and SON cell cultures (P = 0.0001-0.0060). The phenotypically identified cells, cultured in the 300 mOsm/l medium, responded by depolarization or hyperpolarization when transferred to hypertonic or hypotonic perfusion salines, respectively. FAU - Kusano, K AU - Kusano K AD - Laboratory of Neurochemistry, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA. FAU - House, S B AU - House SB FAU - Gainer, H AU - Gainer H LA - eng PT - Journal Article PL - United States TA - J Neuroendocrinol JT - Journal of neuroendocrinology JID - 8913461 RN - 0 (Antibodies) RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Hypotonic Solutions) RN - 11000-17-2 (Vasopressins) RN - 451W47IQ8X (Sodium Chloride) RN - 50-56-6 (Oxytocin) SB - IM MH - Action Potentials/drug effects MH - Animals MH - Antibodies MH - Brain-Derived Neurotrophic Factor/*pharmacology MH - Cell Survival/drug effects MH - Cells, Cultured MH - Electrophysiology MH - Female MH - Fluorescent Antibody Technique MH - Hypothalamus/*cytology MH - Hypotonic Solutions/pharmacology MH - Immunophenotyping MH - Male MH - Neurons/*cytology/drug effects/physiology MH - Osmotic Pressure MH - Oxytocin/analysis/immunology/*physiology MH - Rats MH - Rats, Sprague-Dawley MH - Sodium Chloride/pharmacology MH - Vasopressins/analysis/immunology/*physiology MH - Water-Electrolyte Balance/physiology EDAT- 1999/02/27 00:00 MHDA- 1999/02/27 00:01 CRDT- 1999/02/27 00:00 PHST- 1999/02/27 00:00 [pubmed] PHST- 1999/02/27 00:01 [medline] PHST- 1999/02/27 00:00 [entrez] AID - 10.1046/j.1365-2826.1999.00296.x [doi] PST - ppublish SO - J Neuroendocrinol. 1999 Feb;11(2):145-52. doi: 10.1046/j.1365-2826.1999.00296.x.