Figure 6.
GnRH, vGluT2, and GPR54 mRNA colocalize in KiSS-1-activated MSDB vGluT2-GFP neurons. KiSS-1-activated vGluT2-GFP MSDB neurons were identified using cell-attached recordings, after which the cell cytoplasm was harvested for single-cell RT-PCR using the whole-cell mode of patch clamping. a1, a2, Cell-attached recordings show two KiSS-1-activated vGluT2-GFP neurons from which cytoplasm was harvested for mRNA analysis. a3, A different neuron is shown in each of the 11 lanes. Cells in a1 and a2 are shown in lanes 2 and 5. To demonstrate a successful harvest of a cell, β-actin was used. Note that 9 of 11 vGluT2-GFP cells shown expressed vGluT2 mRNA. GnRH mRNA expression was observed in 5 of 5 cells shown. One control lane was used with water (H20) instead of cell contents; as expected, this lane was negative. b, For control purposes, we also harvested neurons from the neighboring striatum, where no vGluT2 or GnRH is expected. All six cells expressed β-actin, but none of the cells showed GnRH or vGluT2 expression. MW, Molecular weight.