第16回千葉カルシウム代謝研究会

Gene ontology term enrichments for RNA-Seq data from differentiated TSC2 deletion cell lines and microarray data of patient SEGAs (related to Fig. 2f). (XLSX 27.7 kb

14-day voluntary running wheel experiment.

<p>In two separate experiments, adult mice with or without access to a running wheel were sacrificed after 2 weeks. The right hemisphere was dissected for either protein or mRNA-analysis while the left hemisphere was used to confirm exercise-induced increase in Dcx-IR via immunohistochemistry. A, Dcx-IR was quantified by calculating the total Dcx-IR area in µm² for four different sections within the dorsal hippocampus (left). Bar graph of hippocampal Dcx-protein-levels in DG and resHp (right). N = 12/group. B, Dcx-IR was quantified by calculating the total Dcx-IR area in µm² for four different sections within the dorsal hippocampus (left). Bar graph of hippocampal Dcx-mRNA-level in DG and resHp (right). N = 15/group. C, Bar graph mRNA-level in DG and resHp for Sox11 and Tbr2/EOMES. N = 15/group. Bonferroni’s Multiple Comparisons Test.</p

Dcx protein-levels in rat brain homogenates and CSF during development.

<p>Olfactory bulb, whole hippocampus, pieces of cerebral cortex and cerebellum, and CSF were analyzed from rats at different developmental stages (postnatal day 5 to 40). A, Dcx-protein levels in rat brain tissue homogenates during postnatal development. B, Dcx-protein levels in CSF during postnatal development (N = 4).</p

Irradiation-induced ablation of neurogenesis.

<p>A, schematic diagram of experimental procedures. Female wistar rats received a high (12 gy) or low (6 gy) irradiation dose or were sham-treated at P10 (N = 10 per group). Mice were sacrificed 7 weeks after treatment. A subset of brains were processed for immunohistochemistry (12 gy: N = 2, 6 gy: N = 3, sham: N = 3). Residual brains were split into hemispheres and dissected for mRNA and protein analysis (N = 6/group). B, Representative images of Dcx-IR in the olfactory bulb of formalin-fixed paraffin embedded (FFPE) sections. Left-to-right: A high density of dendritic Dcx-IR is observed in the olfactory bulb granular layer sham-irradiated animals. Dendritic labeling is reduced with low-irradiation and virtually absent in animals after 12 gy-irradiation. Scale bar: 200 µm. C, bar graphs of Dcx-protein-levels in the olfactory bulb. A dose-dependent decrease in Dcx-protein levels is observed in irradiated animals vs sham-controls. D, bar graphs of Dcx-mRNA-levels in the olfactory bulb. A dose-dependent decrease in Dcx-mRNA levels is observed in irradiated animals vs sham-controls. E, Representative images Dcx-IR in the dentate gyrus of FFPE sections. Upper panel: overview of Dcx-IR in the dentate gyrus. Left-to-right: Dcx-IR is restricted to cells in the dentate gyrus SGZ with dendrites spanning into the granular and molecular layer. Lower panel: higher magnification of the SGZ. F, bar graphs of Dcx-protein-levels in the hippocampus. A slight dose-dependent decrease in DCX-protein levels is observed in irradiated animals. G, bar graphs of Dcx-mRNA-levels in the hippocampus. DCX-mRNA levels do not change significantly between sham and irradiated groups. H, bar graphs of Dcx-protein-levels in the cerebral cortex. I, bar graphs Dcx-mRNA-levels in cerebral cortex. DCX-mRNA levels do not change significantly between sham and irradiated groups. J, bar graphs of Dcx-protein-levels in the cerebellum. K, bar graphs of Dcx-mRNA-levels in cerebellum. Dcx-mRNA levels do not change significantly between sham and irradiated groups. Dunnett’s Multiple Comparisons Test.</p

Murine Dcx-protein expression using a Dcx-specific immunoassay.

<p>A, Left, Dcx is detected in whole brain homogenates from adult C57BL/6 wt and Dcx-KO mice using a sandwich immunoassay. No signal can be observed in Dcx-KO mice (N = 3). Right, Representative Dcx immunoblot. Dcx can be detected in hippocampal tissue of adult wildtype but not Dcx KO mice. B, expression levels of Dcx-protein in various mouse brain regions (wHp: whole hippocampus, Cx: cortex, Cb: Cerebellum, OB: olfactory bulb, N = 6).</p

Additional file 10: Table S7. of Genomic analysis of the molecular neuropathology of tuberous sclerosis using a human stem cell model

RNA-Seq and ribosome profiling data of differentiated TSC2 wild-type and homozygous deletion cell lines treated with mTOR inhibitors (related to Fig. 4). (XLSB 7.52 mb

Additional file 11: Table S8. of Genomic analysis of the molecular neuropathology of tuberous sclerosis using a human stem cell model

Gene sets and associated genes that show after mTOR treatment a reversal of the change in expression detected in untreated TSC2 deletion cells (related to Fig. 4b). (XLSX 25.8 kb

Additional file 8: Table S5. of Genomic analysis of the molecular neuropathology of tuberous sclerosis using a human stem cell model

Gene set enrichment analysis of ribosome profiling data (related to Fig. 3d). (XLSX 11.1 kb

Additional file 2: Figure S2. of Genomic analysis of the molecular neuropathology of tuberous sclerosis using a human stem cell model

Loss of TSC2 triggers expression changes related to inflammatory response, metabolism, and neuronal function. (PDF 1.15 mb

Additional file 1: Figure S1. of Genomic analysis of the molecular neuropathology of tuberous sclerosis using a human stem cell model

Deregulated expression of neuronal and glial markers in the absence of TSC2. (PDF 2.73 mb