Abstract
ObjectTo explore the mechanisms of ovarian aging, we performed overall analysis on the age-related alterations of gene expression profiles in mouse germinal vesicle (GV) stage oocytes by means of single-cell RNA-sequencing method (scRNA-seq).MethodsTwo age groups (5-week-old and 32-week-old) female KM mice were used as young and old models. Subsequently, GV oocytes were collected for scRNA-seq. The bioinformatics was performed to analyze and compare the differences of gene expression profile between GV oocytes of young and old mice.ResultsThe analysis of scRNA-seq data showed that there were 624 differential expressed genes (DEGs) between two age groups of mouse GV stage oocytes. Four hundred forty-nine DEGs were up-regulated while 175 DEGs were down-regulated in the GV oocytes of the old group. KEGG pathway analysis revealed that the genes involved in mitochondrial function including oxidative phosphorylation and ATP production pathway were significantly down-regulated in GV oocytes of 32-week-old mice, especially the mitochondrial encoded NADH dehydrogenase (mt-Nd), including mt-Nd2, mt-Nd3, mt-Nd4, mt-Nd4L and mt-Nd5. Analysis of DEGs revealed that endoplasmic reticulum stress-related genes including AdipoR2, IRAK-1, RCAN1 and MsrB1 were significantly down-regulated in GV oocytes of 32-week-old mice. Also, analysis of DEGs demonstrated that anti-oxidation-related genes including Erbb3、Rcan1、Gsto2 and Msrb1 were significantly down-regulated in GV oocytes of old group.ConclusionThe disorder of mitochondrial function, endoplasmic reticulum stress and the reduced antioxidant capability might be involved in the progression of oocyte aging. Especially, the down regulation of mitochondrial encoded subunits of respiratory chain complexes might play critical roles in the relevant mechanisms.
Highlights
In modern society, more and more females choose to delay reproduction to pursue educational and career goals
Boxplots of log10-transformed FPKM values for each samples revealed that these samples had consistent overall range and distribution of the FPKM values (Fig. 1a), indicating the single-cell RNA-sequencing data of this study was of reliability, reproducibility and high quality
Cluster analysis of gene expression levels (FPKM) in each sample showed the differences of gene expression profile between GV oocytes of 5-week-old and 32-weekold mice, which revealed that the RNA-sequencing data of this study met the conditions for differential expression analysis (Fig. 1b)
Summary
Two age groups (5-week-old and 32-week-old) female KM mice were used as young and old models. The bioinformatics was performed to analyze and compare the differences of gene expression profile between GV oocytes of young and old mice
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