Lrrc34 Is Highly Expressed in SSCs and Is Necessary for SSC Expansion In Vitro.
- Author:
Jin Huan OU
1
;
Yi Ran LI
1
;
Zhi Peng WANG
1
;
Cheng JIN
1
;
Kai LI
1
;
Yan LU
1
;
Ding Feng ZOU
1
;
Peng Yu LI
1
;
Meng Zhen LI
1
;
Shi Ying MIAO
1
;
Lin Fang WANG
1
;
Wei SONG
1
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis/genetics*; Cell Proliferation/genetics*; Cells, Cultured; Gene Expression Profiling/methods*; Gene Ontology; Gene Regulatory Networks; Humans; Male; Mice, Inbred C57BL; Mice, Transgenic; RNA Interference; Repressor Proteins/metabolism*; Signal Transduction/genetics*; Stem Cells/metabolism*
- From: Chinese Medical Sciences Journal 2020;35(1):20-30
- CountryChina
- Language:English
- Abstract: Objective To discover critical genes contributing to the stemness and maintenance of spermatogonial stem cells (SSCs) and provide new insights into the function of the leucine-rich repeat (LRR) family member Lrrc34 (leucine-rich repeat-containing 34) in SSCs from mice. Methods Bioinformatic methods, including differentially expressed gene (DEG), gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were used to uncover latent pluripotency-related genes. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels, respectively. RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs. Results Eight DEGs between ID4-EGFP+ (G) and ID4-EGFP+/TSPAN8High (TH), eight DEGs between G and ID4-EGFP+/TSPAN8Low (TL) and eleven DEGs between TH and TL were discovered, and eleven protein-protein interaction (PPI) modules were found to be significant in the PPI network of DEGs. One of the DEGs, Lrrc34, was selected as a potential pluripotency-related gene due to its differential expression among ID4-EGFP+ spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module. Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A, and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice. The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways. Conclusion Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways.
