Subcellular distribution of alpha1 and beta2/3 GABA(A) receptor subunits in sensory neurons of the bovine trigeminal mesencephalic nucleus: evidence suggesting their axoplasmic transport.

Neurons were free-hand isolated under the stereomicroscope from the bovine trigeminal mesencephalic nucleus. These neurons were challenged with monoclonal antibodies against the alpha1 and the beta2/3 subunits of the GABA(A) receptor. The neurons showed a strong reaction to both antibodies. The reaction was mainly intracellular in the round cell bodies and in the axoplasm of the axon emerging from these pseudounipolar cells. This suggests the synthesis of these subunits in the cell body and their transport along the axon.

Immunocytochemical study of alpha 1 and beta 2/3 subunits of GABAA receptors in freehand isolated vestibular Deiters' neurons.

Vestibular Deiters' neurons have been isolated from bovine brain by the Hydén's freehand dissection technique and challenged with monoclonal antibodies directed toward the alpha 1 and beta 2/3 subunits of the GABAA receptors. Subsequent challenge with fluorescent secondary antibodies and confocal microscopy allowed the study of the cellular distribution of such subunits. In Deiters' neurons the beta 2/3 subunit displayed a clear presence all along the cell body profile and the initial parts of the dendrites. The alpha 1 subunit was found highly present all over the cell interior except the nuclear profiles. The strong presence inside the cells possibly masked its presence on the plasma membrane. However, in part of the cells studied a distinct presence on the plasma membrane was evident. This subunit was visualized also all along the long dendrites of these neurons. The approach we describe here, involving freehand isolated mature neurons from adult animals, may allow a better characterization of the tridimensional distribution of different types of neuronal GABAA receptors in the respect of the approach with brain slices.