Fig. 4.

Lentiviral expression of PcdhγC4 rescues cINs with Pcdhγ loss of function. (A) Diagram of mouse genetic crosses. Pcdhγ^fcon3 mice were crossed to the Nkx2.1^Cre;Ai14 mouse line to generate embryos with loss of function of all Pcdhγ isoforms. (B) Schematic of the lentiviral infection and transplantation of MGE cIN precursors. The MGEs of Nkx2.1^Cre;Ai14;Pcdhγ^fcon3/fcon3 embryos were dissected, dissociated, and infected in suspension with lentivirus carrying PcdhγC4-GFP. The mixture of transduced and nontransduced cells was grafted into the cortex of WT neonate recipient mice. (C) Confocal images of the transplanted cINs in the cortex at 6 and 21 DAT. Notice the expression of GFP can be found near the cell surface including the cell processes, reflecting the putative location of the transduced PcdhγC4-GFP protein. Quantifications of the tdTomato⁺GFP⁻ (red cells, teal arrows) or tdTomato⁺GFP⁺ (yellow cells, white arrows) cells are shown as the fraction of cells from the total tdTomato⁺ cells at 6 and 21 DAT. The fraction of the PcdhγC4-transduced yellow cells increases from 6 to 21 DAT, while the fraction of nontransduced (tdTomato⁺ GFP⁻) decreases at equivalent time points. (D) Survival fraction quantifications from (C) shown by individual brain sections (each bar) and separated by animals at 6 and 21 DAT. Scale bar = 50 μm, nested ANOVA, ****P = 0.0002, n = 4 mice per time point and 10 brain sections per mouse from one transplant cohort. DAT 6 cell counted = 15,071, DAT 21 cells counted = 1,660.