Culturing embryonic nasal explants for developmental and physiological study.
ABSTRACT Primary cultures obtained from embryonic nasal placodes can maintain olfactory neurons, olfactory ensheathing cells, and large numbers of gonadotropin releasing hormone-1 (GnRH) neurons. Depending on the age of the starting material, one can examine cell interactions important for placode formation or neuronal migration and axonal outgrowth. When generated at E11.5 in mouse, neuronal migration and axon outgrowth away from the main tissue mass occurs. This area of the explant, the periphery, is only a few cells thick. This characteristic offers the opportunity to image single cells and axons and allows pharmacological and molecular manipulations as well as physiological recordings to be performed. Here, we describe a system for culturing nasal explants used in our laboratory. This model system provides a method for obtaining physiological cellular responses with post hoc immunohistochemistry, or gene expression studies, on cells arising from the nasal placode.
[show abstract] [hide abstract]
ABSTRACT: Many types of neurons and glia exhibit oscillatory changes in membrane potentials and cytoplasmic Ca2+ concentrations. In neurons and neuroendocrine cells an elevation of intracellular Ca2+ concentration is associated with neurosecretion. Since both oscillatory membrane potentials and intracellular Ca2+ oscillations have been described in primary LHRH neurons and in GT1 cells, it is evident that an endogenous pulse-generator/oscillator is present in the LHRH neuron in vitro. The hourly rhythms of LHRH neurosecretion appear to be the synchronization of a population of LHRH neurons. How a network of LHRH neurons synchronizes their activity, i.e., whether by the result of synaptic mechanisms or electrical coupling through gap junctions or through a diffusible substance(s), remains to be clarified. Even though LHRH neurons themselves possess an endogenous pulse-generating mechanism, they may be controlled by other neuronal and nonneuronal elements in vivo. NE, NPY, glutamate, and GABA are neurotransmitters possibly controlling pulsatile LHRH release, and NO, cAMP, and ATP may be diffusible substances involved in pulsatile LHRH release without synaptic input. Although synaptic inputs to the perikarya of LHRH neurons could control the activity of LHRH neurons, a line of evidence suggests that direct neuronal and nonneuronal inputs, especially those from astrocytes to LHRH neuroterminals, appear to be more important for pusatile LHRH release.Vitamins & Hormones 02/2001; 63:91-129. · 2.19 Impact Factor