GABAA receptor subunit expression and assembly in cultured rat cerebellar granule neurons

The assembly of multisubunit GABAA receptors in specific neuronal populations is a complex process which is poorly understood. To begin to examine receptor assembly, alpha 1, beta 2/3, and gamma 2 subunit polypeptide expression and association, as well as receptor binding, were examined in cultured rat cerebellar granule neurons. Western blots revealed two alpha 1-immunoreactive proteins. A 39 kDa species was maximal at 2 days in culture and subsequently declined. In contrast, a 51 kDa polypeptide, the anticipated size of the mature alpha 1 subunit, was first detected at 4 days and increased throughout the culture period. Additional studies demonstrated that the beta 2/3 and gamma 2 subunits were detectable at 2 days and attained maximal levels by 6 days. The level of [3H]Ro15-1788 binding, a measure of assembled receptors, rose in parallel with the increases in the 51 kDa alpha 1, beta 2/3 and gamma 2 subunits. Moreover, the 51 kDa alpha 1, beta 2/3, and gamma 2 subunits were associated in receptor complexes. However, immunohistochemical studies demonstrated the presence of substantial intracellular subunit staining. This finding suggest that only some of the subunits expressed in granule neurons contribute to functional GABAA receptors on the cell surface.