Transient expression of GABAA receptor subunit mRNAs in the cellular processes of cultured cortical neurons and glia.
In this study, we have studied by in situ hybridisation histochemistry the expression and intracellular distribution of the GABAA receptor subunit mRNAs in cultured neurons obtained from postnatal day 1-3 rats in order to determine how neurotransmitter receptor expression may be regulated during development of the nervous system. In postnatal cortical cells, we found that GABAA receptor subunit mRNAs coding for alpha2, alpha5, beta2, beta3 and gamma2 subunits were transiently expressed in the cellular processes and growth cones after 1-3 days in culture. These observations indicate that GABAA receptor subunit mRNAs are transported (or trafficked) into the cellular processes of early postnatal cortical cells. These selective localisations were rarely observed after 5 days in culture and only in cells which had not made cell-to-cell contact. The localisation of subunit mRNAs in the processes was more effectively maintained up to 5 days or even longer if cell-to-cell contact was avoided by culturing the cells at low density or by inhibiting neurite sprouting pharmacologically with the GABA receptor channel antagonist TBPS. Finally, immunocytochemistry revealed the expression of GABAA receptors in the growth cones of pyramidal neurons in culture. Thus, the expression of mRNA correlates to the expression of protein. These results suggest that the selective trafficking of GABAA receptor subunit mRNAs during synaptogenesis may be regulated by synapse formation and/or glial-neural communication.
Poulter
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Laboratory of Molecular Neuropharmacology, Institute of Biological Sciences, National Research Council of Canada, Bldg. M-54, Montreal Road Campus, Ottawa, Ontario, Canada. mike.poulter@nrc.ca
Brown
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D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D005260:Female; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D020439:Growth Cones / Q000737:chemistry / Q000502:physiology; D017403:In Situ Hybridization; D009457:Neuroglia / Q000737:chemistry* / Q000166:cytology / Q000502:physiology; D017966:Pyramidal Cells / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000032:analysis / Q000235:genetics*
D012333:RNA, Messenger; D011963:Receptors, GABA-A