Comment[GEAAccession]	E-GEAD-285
MAGE-TAB Version	1.1
Investigation Title	Aging of mouse spermatogonial stem cells by Wnt7b-Jnk pathway activation
Experiment Description	Although spermatogonial stem cells (SSCs) are thought to be virtually immortal, the number of SSCs declines during aging, which is considered to be induced by deterioration of microenvironment. Here we report that cell-intrinsic mode of SSC aging. SSCs cultured for 5 years proliferate more actively than young SSCs and showed enhanced glycolytic activity but remain euploid and exhibit stable androgenetic imprinting patterns. Moreover, the aged SSCs kept constant SSC activity despite shortened telomeres. The increased proliferative activity of aged SSCs was caused by Wnt7b expression, which likely occurred due to decreased polycomb complex2 activity. Aberrant Wnt7b expression not only decreased reactive oxygen species levels and mitochondria activity but also stimulated glycolysis via JNK activation. Analysis of SSCs in Klotho aging mouse model also confirmed hyperactivation of JNK pathway and increased glycolysis. Therefore, not only SSC microenvironment but also intrinsic activation of Wnt7b-mediated glycolysis plays important roles in SSC aging.
Experimental Design	cell type comparison design	replicate design
Experimental Factor Name	culture_duration
Experimental Factor Type	culture_duration
Person Last Name	Iwama	Oshima	Shinohara	Shinohara
Person First Name	Atsushi	Motohiko	Mito	Takashi
Person Affiliation	Cellular Molecular Biology, Translational Medicine, Graduate School of Medicine, Chiba University			
Person Roles	submitter	submitter	submitter	submitter
Public Release Date	2022-10-15
Protocol Name	P-GEAD-34	P-GEAD-35	P-GEAD-36	P-GEAD-37	P-GEAD-38
Protocol Type	sample collection protocol	nucleic acid extraction protocol	nucleic acid library construction protocol	nucleic acid sequencing protocol	normalization data transformation protocol
Protocol Description	Harvest culture cells.	Total RNA was extracted from GS cells cells using RNeasy Plus Micro Kit (Qiagen, Valencia, CA).	cDNA libraries were generated using a NEBNext Ultra RNA Library Prep Kit (New England BioLabs, Beverly, MA).	Sequencing was performed using HiSeq1500 (Illumina) with a single-read sequencing length of 60bp.	TopHat (version 2.0.13; with default parameters) was used to map to the reference genome (UCSC/mm10 or UCSC/hg19) with annotation data from iGenomes (Illumina). Then, levels of gene expression were quantified using Cuffdiff (Cufflinks version 2.2.1; with default parameters).
SDRF File	E-GEAD-285.sdrf.txt
Comment[AEExperimentType]	RNA-seq of coding RNA
Comment[BioProject]	PRJDB7461
Comment[Last Update Date]	2022-10-15