Expression of Neurotransmitter Receptors by mRNAs from Neurons Developing In Vitro: A Xenopus Oocyte Expression Study

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Abstract
Poly(A)+ mRNA was extracted from cultures of neurons isolated from mouse embryonic day 14 cerebral cortex and injected into Xenopus oocytes. This led to the expression of receptors for gamma-aminobutyric acid (GABA), glycine, acetylcholine, serotonin, glutamate, kainate, N-methyl-D-aspartate, and quisqualate. Northern blot analysis of poly(A)+ mRNA from the cultured neurons with a GluR1 cDNA probe revealed the presence of three hybridization bands with estimated mRNA sizes of 5.1, 4.0, and 3.1 kb, respectively. The development of mRNAs coding for neurotransmitter receptors was investigated by isolating mRNA from neurons cultured for 2, 8, and 14 days in vitro and injecting it into Xenopus oocytes. The amplitude of membrane currents elicited by the transmitters gave a measure of the relative amounts of the different mRNAs. The size of the responses to kainate, aspartate (together with glycine), glutamate, acetylcholine, GABA, serotonin, and glycine increased with the time of culture in vitro. However, in contrast to all other agonist-induced currents, the current induced by glycine failed to increase further from 8 to 14 days in culture. It is concluded that the time course of receptor development in cortical neurons in vitro is similar to the development in vivo.