Figure 8.

Representative SSTR4 and VGLUT1 mRNA expression of layer V pyramidal neurons of human neurosurgical cortical samples taken from the left middle temporal gyrus (by multiplex fluorescent RNAscope in situ hybridization (ISH)). Panel (A) shows images from Sample A of a 56-year-old man. Note that the tissue contains considerable amount of lipofuscin that shows some autofluorescence in all examined channels (arrows). Granular fluorescence pattern that occurs in one color only represents the respective specific RNAscope ISH signal for VGLUT1 (green) or SSTR4 (red, marked in part by arrowheads) counterstained with DAPI (blue). Panel (B) depicts images of Sample B taken from a 19-year-old woman. Note the relatively low aging pigment content (arrows) of the VGLUT1 (green) expressing layer V pyramidal cells, which contain SSTR4 mRNA (arrowheads) also, counterstained with DAPI (blue). Cellular and nuclear (DAPI) morphology in Sample B differs from that of Sample A, as Sample B was fresh frozen before fixation, in contrast to Sample A that was subjected to immediate immersion fixation in 10% neutral buffered formalin. Boxed areas in the top images (scale bar: 100 µm) of both panels are shown in higher magnification photos right bellow (scale bar: 25 µm).