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. 2009 May 6;29(18):5806–5819. doi: 10.1523/JNEUROSCI.1106-09.2009

Figure 2.

Figure 2.

Distribution and quantification of neurons expressing the ChR2-mCherry transgene. A, Distribution of C1 (ChR2+TH+, yellow squares) and noncatecholaminergic neurons (ChR2+TH−, red circles) expressing ChR2-mCherry in a representative rat (neuronal nuclear profiles counted in a 1-in-6 series of 30-μm-thick transverse sections through the injection site). Nontransfected C1 neurons (ChR2−TH+, blue triangles) are also represented. Each symbol represents one cell. The numbers in the top left corner of each section refer to the distance of the section behind bregma in millimeters [after the atlas of Paxinos and Watson (2005)]. By our convention, 11.6 mm is designated as the first section in which the facial motor neurons appear from a caudal approach, and the rest of the sections are placed relative to this landmark. Brain reconstructions were made using the Neurolucida software (MBFBioscience) and a Zeiss Axioskop II microscope with epifluorescence using filter sets for Cy3 and Alexa 488 fluorophores. Note the vertical lesion caused by the placement of the fiber optic in the section corresponding to bregma level 11.42. B, Group data. Two of the plots show the rostrocaudal distribution of noncatecholaminergic (TH−, red circles, red line) and catecholaminergic (TH+, yellow squares, black line) neurons that expressed mCherry after injection of PRSX8-ChR2-mCherry virus into the RTN region (mean ± SEM; 7 rats). The remaining plot shows the distribution of all the neurons (TH+ and TH−) that expressed eGFP after injection of PRSX8-Allatostatin receptor-IRES-eGFP virus into the RTN region (black circles, dashed line; 5 rats). C, Total number of counted catecholaminergic (TH+) and noncatecholaminergic neurons expressing the transgene after injection of PRSX8-ChR2-mCherry lentivirus (7 rats; cell counts in black bars) or PRSX8-AllatoR-IRES-EGFP lentivirus (5 rats; cell counts of EGFP-ir in gray bars) into the RTN region (mean ± SEM). Cell counts were made in each rat in a one-in-six series of 30 μm sections as in A. Accordingly, the estimated total number of transfected neurons is approximately six times that reported on the graph.