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. 2023 Oct 16;10(10):ENEURO.0054-23.2023. doi: 10.1523/ENEURO.0054-23.2023

Figure 6.

Figure 6.

Neonatal intraventricular injection of adeno-associated virus results in lasting and highly specific transgene expression in Cajal–Retzius cells. A, Representative images of a ΔNp73-Cre mouse hippocampus 7 weeks post-neonatal (P0) injection with AAV8/EF1a-DIO-ChR2-mCherry. Coimmunostaining with TRP73 and RELN indicates CR cells continue to express mCherry 7 weeks following the injection (yellow arrows). Some CR cells do not express mCherry (white arrowhead) because they were not transduced by the virus. Scale bars: top, 200 μm; bottom, 20 μm. B, Quantification of AAV-transduced CR cells (mCherry+ TRP73+) in 7-week-old wild-type (WT) and ΔNp73-Cre mice. C, Comparison of the number of mCherry+ granule neurons in P14 and 7-week-old ΔNp73-Cre mice neonatally injected with AAV8/EF1a-DIO-ChR2-mCherry. D, Comparison of the number of recombination events in granule neurons and CR cell-targeting specificity of 7-week-old ΔNp73-Cre; LSL-tdTomato mice, ΔNp73-Cre mice neonatally injected with AAV8/EF1a-DIO-ChR2-mCherry, and ΔNp73-Cre mice neonatally injected with AAV8/hSyn-DIO-hM3D-mCherry. E, Representative images of the hippocampal fissure areas of P14 and 7-week-old wild-type and ΔNp73-Cre transgenic mice injected neonatally with AAV8/EF1a-DIO-ChR2-mCherry. Scale bars, 20 μm. Quantification of the densities of TRP73+ CR cells are shown at the bottom. SLM, Stratum lacunosum-moleculare; HF, hippocampal fissure; ML, molecular layer; GL, granular layer; CA1, Cornu Ammonis 1; loML, lower molecular layer (i.e., molecular layer of the infrapyramidal blade of the dentate gyrus). Data are presented as scatter plots in B, C, and D with all data points shown and error bars representing ±SD, and statistical analyses were performed using nested one-way ANOVA with Tukey’s post hoc test. Each data point is an individual animal, whereby three sections were measured for each animal. Data are presented as summary data in E with mean ± SD, and statistical analyses were performed using two-way ANOVA with Sidak test to correct for multiple comparisons. ***p < 0.001; ****p < 0.0001; n.s., not significant.