Serum-free culture of rat postnatal neurons derived from the dorsal motor nucleus of the vagus

Previous studies on dorsal motor nucleus of the vagus (DMNV) neurons have mainly used in vivo animal models and in vitro brainstem slices. Primary culture of postnatal DMNV neurons in defined serum free medium has not been reported. We report a method for culture of postnatal rat DMNV neurons using serum free medium. Cultured DMNV neurons contain both Hu positive precursor cells and mature cells staining positively for microtubule associated protein 2 (MAP2) and choline acetyltransferase. Exposure of cultured DMNV neurons to glutamate (10(-7) to 10(-3) M) induced an increase in intracellular calcium concentration ([Ca2+](i)) in a dose-dependent manner, indicating the functional presence of glutamate receptors. Voltage-dependent calcium currents were present in cultured DMNV neurons. Active cell proliferation was demonstrated by BrdU incorporation. Upon removal of beta FGF, the percentage of MAP2 positive mature neurons was significantly increased from 36 +/- 3 to 73 +/- 3%. Our study demonstrates that postnatal rat DMNV neurons cultured in serum free medium retain morphological and physiological characteristics of DMNV neurons in situ. (c) 2005 Elsevier B.V. All rights reserved.