Interruptions of dentate gyrus granule cell neurogenesis induced by sevoflurane exposures in male and female mice. (A) The t-distributed stochastic neighbor embedding plots showing nine dentate gyrus granule cell subclusters in the control male group (left) and sevoflurane male group (right). (B) The percentage of cells in specific dentate gyrus granule subclusters in the control male and sevoflurane male groups. (C) Heatmap showing the expression of specific marker genes for dentate gyrus granule subclusters. Criteria for specific marker genes: mincell_pct 0.25 or greater, P value ≤ 0.01, and log₂(fold change) 0.361 or greater (Seurat R package). (D) Heatmap showing the expression of specific marker genes (rows) in subclusters (columns) along the maturation trajectory from neural stem cells to neural progenitors. (E) Pseudotime expression graphs of representative specific marker genes (Slc1a3, Sox6, and Neurod2) showing the development profile of dentate gyrus granule cells in the control male group (upper) and sevoflurane male group (lower). Color intensity corresponds to the states of neurogenesis. (F) Interrupted expression of Slc1a3 [F(6,680) = 4,074, P < 0.001], Sox6 [F(6,453) = 18,551, P < 0.001], and Neurod2 [F(6,453) = 2,576, P < 0.001] along the pseudotime course. Data are mean ± SD. n = 863 cells (control male) and n = 2,288 cells (sevoflurane male). Mixed-effect ANOVA with Bonferroni correction. ***P < 0.001. (G) The t-distributed stochastic neighbor embedding plots showing 16 dentate gyrus granule cell subclusters in the control female group (left) and sevoflurane female group (right). (H) The percentage of cells in specific dentate gyrus granule subclusters in the control female and sevoflurane female groups. (I) Heatmap showing the expression of specific marker genes for dentate gyrus granule cell subclusters. Criteria for specific marker genes: mincell_pct 0.25 or greater, P value ≤ 0.01, and log₂(fold change) 0.361 or greater (Seurat R package). (J) Heatmap showing the expression of specific marker genes (rows) in subclusters (columns) along the maturation trajectory from neural stem cells to neural progenitors. (K) Pseudotime expression graphs of specific marker genes showing the development profile of dentate gyrus granule cells in the control female group (left) and sevoflurane female group (right). Color intensity corresponds to the states of neurogenesis. (L) Interrupted expression of Dcx [F(3,35) = 400.2, P < 0.001], Draxin [F(3,33) = 15.1, P < 0.001], and Fabp7 [F(3,62) = 33,779, P < 0.001] along the pseudotime course. Data are mean ± SD. n = 6,894 cells (control female) and n = 6,676 cells (sevoflurane female). Mixed-effect ANOVA with Bonferroni correction. ***P < 0.001.