Abstract
Trigeminal mesencephalic nucleus neurons transmit oral sensation. The soma of these neurons contain GABA(A) receptors. Our objective is to quantify the gene expression corresponding to subunits of the GABA(A) receptor in trigeminal neurons in the mesencephalic nucleus, particularly those that project to muscle spindles of the masseter muscle. In order to accomplish this objective, we have developed a protocol in which we identify the target soma using retrograde fluorescence staining, resect and collect them with a laser-microdissection system, and quantify the expression of the genes coding for subunits of the GABA(A) receptor using PCR. We have tested this protocol with the following results :1.The retrograde fluorescence-staining technique using dextran-rhodamine B efficiently identifies the target soma to be resected with the laser-microdissection system.2.Resection with a laser-microdissection system is an efficient means to collect micro-structural tissue samples.3.Consistent with the report of Ishii et al, we detected the mRNA of each of the subunits of the GABA(A) receptor in soma resected and collected using the laser-microdissection system.
