Characterization of sheep spermatogenesis through single‐cell RNA sequencing

Yang, Hua; Ma, Junwu; Wan, Zhen; Wang, Qi; Zhao, Jie; Wang, Zhen; Zhang, Yanli

doi:10.1096/fj.202001035rrr

Cited by 36 publications

(42 citation statements)

References 43 publications

(44 reference statements)

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“…By knowing the changes in the transcription level of various cell types during the process of sheep spermatogenesis, Researchers found that the marker genes of early and late primary S’cytes were primarily involved in the cell cycle and related to meiosis. The other three stages were mainly related to sperm deformation such as being involved in metabolism, spermatogenesis, sperm development, and differentiation ( Yang et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Identification of Key Genes Related to Postnatal Testicular Development Based on Transcriptomic Data of Testis in Hu Sheep

Sun

Pei

et al. 2022

Front. Genet.

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The selection of testis size can improve the reproductive capacity of livestock used for artificial insemination and has been considered as an important strategy for accelerating the breeding process. Although much work has been done to investigate the mechanisms of testis development in various species, there is little information available in regard to the differences in transcriptomic profiling of sheep testes at different developmental stages. In this work, we aimed to identify differentially expressed genes (DEGs) by RNA-Seq in sheep during different growth stages, including 0 month old (infant, M0), 3 months old (puberty, M3), 6 months old (sexual maturity, M6) and 12 months old (body maturity, M12). A total of 4,606 (2,381 up and 2,225 down), 7,500 (4,368 up and 3,132 down), 15 (8 up and seven down) DEGs were identified in M3_vs_M0, M6_vs_M3, and M12_vs_M6 comparison, respectively. Of which, a number of genes were continuously up-regulated and down-regulated with testicular development, including ODF3, ZPBP1, PKDREJ, MYBL1, PDGFA, IGF1, LH, INSL3, VIM, AMH, INHBA, COL1A1, COL1A2, and INHA. GO analysis illustrated that DEGs were mainly involved in testis development and spermatogenesis. KEGG analysis identified several important pathways and verified several reproduction-associated DEGs such as COL1A1, COL1A2, PDGFA, and IGF1. In addition, two gene modules highly associated with testis development and core genes with testis size were identified using weighted gene co-expression network analysis (WGCNA), including hub genes positively associated with testis size such as RANBP9, DNAH17, SPATA4, CIB4 and SPEM1, and those negatively associated with testis size such as CD81, CSK, PDGFA, VIM, and INHBA. This study comprehensively identified key genes related to sheep testicular development, which may provide potential insights for understanding male fertility and better guide in animal breeding.

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Section: Discussionmentioning

confidence: 99%

Identification of Key Genes Related to Postnatal Testicular Development Based on Transcriptomic Data of Testis in Hu Sheep

Sun

Pei

et al. 2022

Front. Genet.

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“…In the present study, we identified spermatogonia, spermatocytes, spermatids, Sertoli cells, Leydig cells and myoid cells through scRNA-seq. Recently, Yang et al reported characterization of sheep spermatogenesis via scRNA-seq [ 20 ]. However, spermatogonia cluster was not identified in their study.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, the laser capture micro-dissection approach has been used to precisely capture subtypes of male germ cells from paraffin tissue sections, but it leads to RNase contamination after staining [ 15 ]. Fortunately, 10 × Genomics single-cell RNA-sequencing (scRNA-seq) is an unbiased approach with high efficiency and sensitivity in cell capture, and it has been used to decipher the dynamic process of spermatogenesis in mouse, primate and sheep testis [ 16 – 20 ]. However, the gene expression profiling of spermatogenic cells at each sub-stage during spermatogenesis and their regulatory networks in pigs have not been uncovered.…”

Section: Introductionmentioning

confidence: 99%

Single-cell RNA-sequencing reveals the dynamic process and novel markers in porcine spermatogenesis

Zhang

Lei

et al. 2021

J Animal Sci Biotechnol

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Background Spermatogenesis is the process by which male gametes are formed from spermatogonial stem cells and it is essential for the reliable transmission of genetic information between generations. To date, the dynamic transcriptional changes of defined populations of male germ cells in pigs have not been reported. Results To characterize the atlas of porcine spermatogenesis, we profiled the transcriptomes of ~ 16,966 testicular cells from a 150-day-old pig testis through single-cell RNA-sequencing (scRNA-seq). The scRNA-seq analysis identified spermatogonia, spermatocytes, spermatids and three somatic cell types in porcine testes. The functional enrichment analysis demonstrated that these cell types played diverse roles in porcine spermatogenesis. The accuracy of the defined porcine germ cell types was further validated by comparing the data from scRNA-seq with those from bulk RNA-seq. Since we delineated four distinct spermatogonial subsets, we further identified CD99 and PODXL2 as novel cell surface markers for undifferentiated and differentiating spermatogonia, respectively. Conclusions The present study has for the first time analyzed the transcriptome of male germ cells and somatic cells in porcine testes through scRNA-seq. Four subsets of spermatogonia were identified and two novel cell surface markers were discovered, which would be helpful for studies on spermatogonial differentiation in pigs. The datasets offer valuable information on porcine spermatogenesis, and pave the way for identification of key molecular markers involved in development of male germ cells.

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“…Although scRNA-seq technology has been successfully used to construct a map of human spermatogenesis [22][23][24], little research has been performed on the cellular characteristics and regulatory mechanisms of spermatogenesis in domestic animals such as sheep. In only one previous study, the testes of an adult (1.5-year-old) ram were found to contain five germ cell types, two testicular somatic cell types, and an unknown type of somatic cell [25]. However, undifferentiated spermatogonial cells, essential for long-term spermatogenesis, were not captured, and so the underlying mechanisms regulating spermatogonial cell self-renewal/differentiation and spermatogenesis remain unclear.…”

Section: Introductionmentioning

confidence: 99%

The Transcriptional Cell Atlas of Testis Development in Sheep at Pre-Sexual Maturity

Guo

et al. 2022

CIMB

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Sheep testes undergo a dramatic rate of development with structural changes during pre-sexual maturity, including the proliferation and maturation of somatic niche cells and the initiation of spermatogenesis. To explore this complex process, 12,843 testicular cells from three males at pre-sexual maturity (three-month-old) were sequenced using the 10× Genomics ChromiumTM single-cell RNA-seq (scRNA-seq) technology. Nine testicular somatic cell types (Sertoli cells, myoid cells, monocytes, macrophages, Leydig cells, dendritic cells, endothelial cells, smooth muscle cells, and leukocytes) and an unknown cell cluster were observed. In particular, five male germ cell types (including two types of undifferentiated spermatogonia (Apale and Adark), primary spermatocytes, secondary spermatocytes, and sperm cells) were identified. Interestingly, Apale and Adark were found to be two distinct states of undifferentiated spermatogonia. Further analysis identified specific marker genes, including UCHL1, DDX4, SOHLH1, KITLG, and PCNA, in the germ cells at different states of differentiation. The study revealed significant changes in germline stem cells at pre-sexual maturation, paving the way to explore the candidate factors and pathways for the regulation of germ and somatic cells, and to provide us with opportunities for the establishment of livestock stem cell breeding programs.

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Characterization of sheep spermatogenesis through single‐cell RNA sequencing

Cited by 36 publications

References 43 publications

Identification of Key Genes Related to Postnatal Testicular Development Based on Transcriptomic Data of Testis in Hu Sheep

Identification of Key Genes Related to Postnatal Testicular Development Based on Transcriptomic Data of Testis in Hu Sheep

Single-cell RNA-sequencing reveals the dynamic process and novel markers in porcine spermatogenesis

The Transcriptional Cell Atlas of Testis Development in Sheep at Pre-Sexual Maturity

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