KCC2 and PCNA expression are inversely correlated in the forebrain of wild-type fish. (A–C) A representative section from the diencephalon of 3 dpf fish showing PCNA (A) and KCC2 (B) expression, and merged image (C). Symbols indicate the same cells in each image: arrows, PCNA+ cell with high expression near the PVZ; arrowhead, a PCNA+ cell with moderate to low expression at an abventricular position; and asterisk, a PCNA− cell in a perineuropilar position. These cells have no, moderate, and high KCC2 expression, respectively. (D) Line graph of corrected line fluorescence (CLF) for PCNA (green) and KCC2 (red) from a representative transect in the telencephalon, showing the inverse expression relationship between PCNA and KCC2. The transect was drawn from the midline PVZ to the tela choroidea at the outermost aspect of the brain. (E) Scatter plot showing the inverse relationship between PCNA and KCC2 expression in individual wild-type cells. Each set of 3 color coded dots with their connecting line represents a triplet of cells from one specimen. For each sampling, cell 1 is within the abventricular zone with high PCNA expression (A, B, & C, arrow), cell 2 is within the cell-rich parasagittal zone with moderate PCNA expression (A, B, & C, arrowhead), and cell 3 is in the peri-neuropilar zone with low PCNA expression (A, B, & C, asterisk). Scatter indicates the PCNA and KCC2 CFI value within each individual cell represented as PCNA vs. KCC2. n = 16.
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