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The miRNA profiling of goat CD49f-positive and negative testicular cells

ABSTRACT: Purpose: miRNAs, a member of the small RNA, play critical roles in the mammalian spermatogenesis. Spermatogonia was the foundation of spermatogenesis and valuable for the study of spermatogenesis. However, it is still not clear that the expression profiling of the miRNAs in spermatogonia of dairy goat. Methods: The CD49f was one of the surface markers for spermatogonia enrichment by MACS. Therefore, we used CD49f microbeads antibody to purify CD49f-positive and negative cells of dairy goat testicular cells by MACS (Magnetic Activated Cell Sorting), and then in-depth analyzed the miRNA expression in these cells using Illumina sequencing technology. Results: The results of miRNAs expression profiling in purified CD49f-positive and negative testicular cells showed that 933 were miRNAs upregulated in CD49f-positive cells and 916 were miRNAs upregulated in CD49f-negative cells with a 2-fold increase, respectively; some spermatogonial stem cells(SSCs) specific miRNAs and marker genes in testis had a higher level expression in CD49f-positive testicular cells, such as miR-221, miR-23a, miR-29b, miR-24, miR-29a, miR-199b, miR-199a, miR-27a, miR-21. Conclusions: our comparative miRNAome data provided some useful miRNAs profiling data of dairy goat spermatogonia cells and suggested CD49f could be used to enrich dairy goat spermatogonia-like cells, including SSCs.

ORGANISM(S): Capra hircus

PROVIDER: GSE57041 | GEO | 2014/04/25

SECONDARY ACCESSION(S): PRJNA245264

REPOSITORIES: GEO

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The miRNA profiling of goat CD49f-positive and negative testicular cells

Project description:Purpose: miRNAs, a member of the small RNA, play critical roles in the mammalian spermatogenesis. Spermatogonia was the foundation of spermatogenesis and valuable for the study of spermatogenesis. However, it is still not clear that the expression profiling of the miRNAs in spermatogonia of dairy goat. Methods: The CD49f was one of the surface markers for spermatogonia enrichment by MACS. Therefore, we used CD49f microbeads antibody to purify CD49f-positive and negative cells of dairy goat testicular cells by MACS (Magnetic Activated Cell Sorting), and then in-depth analyzed the miRNA expression in these cells using Illumina sequencing technology. Results: The results of miRNAs expression profiling in purified CD49f-positive and negative testicular cells showed that 933 were miRNAs upregulated in CD49f-positive cells and 916 were miRNAs upregulated in CD49f-negative cells with a 2-fold increase, respectively; some spermatogonial stem cells(SSCs) specific miRNAs and marker genes in testis had a higher level expression in CD49f-positive testicular cells, such as miR-221, miR-23a, miR-29b, miR-24, miR-29a, miR-199b, miR-199a, miR-27a, miR-21. Conclusions: our comparative miRNAome data provided some useful miRNAs profiling data of dairy goat spermatogonia cells and suggested CD49f could be used to enrich dairy goat spermatogonia-like cells, including SSCs. miRNA profiles of goat CD49f-positive and negative testicular cells were generated by deep sequencing, in triplicate, using Illumina GAIIx

2014-04-25 | E-GEOD-57041 | biostudies-arrayexpress

Capra hircus

Project description:The miRNA profiling of goat CD49f-positive and negative testicular cells

| PRJNA245264 | ENA

Discovery and Comparative Transcriptome Profiling of Dairy Goat MicroRNAs from Dry Period and Peak Lactation Mammary Gland Tissues

Project description:In this study, two small RNA libraries were constructed using dry period and peak lactation dairy goat mammary gland tissues and sequenced by the Illumina Solexa high-throughput sequencing system. A total of 346 conserved and 95 novel miRNAs were identified in the dairy goat. The expression of miRNAs was confirmed by qRT-PCR in nine tissues and the mammary gland during development cycles. In addition, several candidate miRNAs that may be involved in mammary gland development and lactation were found by the comparison of miRNA expression profiles among different tissue and developmental stages of the mammary gland. This study provides the identification and profile of miRNAs related to the biology of the mammary gland in the dairy goat. The identification of these miRNAs could contribute to understanding the molecular mechanisms of lactation physiology and the development of the mammary gland in the dairy goat.

2014-12-31 | GSE41815 | GEO

An expression atlas of human postnatal testicular cell development

Project description:Postnatal human male gonad development and function are known to involve many genes and pathways but our understanding of genome-wide developmental stage-specific and cell type-specific gene expression is far from complete. Integration of testicular and somatic data could elucidate regulatory mechanisms specifically controlling spermatogenesis and may yield insight into certain reproductive pathologies. Please note: AdMinus means that there is no Ad spermatogonia in the corresponding testicular biopsies of cryptorchid children. AdPlus means that Ad spermatogonia are present in the corresponding testicular biopsies of cryptorchid children. JS refers to Johnsen score.

2012-10-28 | E-TABM-1214 | biostudies-arrayexpress

Transcription profiling of testis biopsies to investigate testis developmental gene expression in cryptorchid boys at risk of azoospermia

Project description:Despite timely and successful surgery, 32% of patients with bilateral and 10% with unilateral cryptorchidism will develop azoospermia. Cryptorchid boys at risk of azoospermia display a typical testicular histology of impaired mini-puberty at the time of the orchidopexy. During mini-puberty increased gonadotropin and testosterone secretion stimulate transformation of gonocytes into Ad spermatogonia. In azoospermia risk group this transformation is to a great extent impaired. This study aimed to analyze data on whole genome expression signatures of undescended testes at risk of developing azoospermia. Twenty-three testicular biopsies from 22 boys were analyzed (19 testes from 18 boys with cryptorchidism) and 4 contralateral descended testes from patients with testicular agenesis. Expression profiling identified 483 genes not or under-expressed in the azoospermia risk group compared with the control and LAZR groups. Annotated loci were associated with spermatogenesis. Other significant genes were cellular defense response genes and hormone controlled loci involved in spermatogenesis. Some genes transcribed in normal adult meiotic and post-meiotic germ cells are activated in healthy juvenile Ad spermatogonia. Thus, molecular events initiating the testicular expression program at the onset of puberty and maintaining it during adulthood occur very early in prepubertal testes. This molecular event is to a great extent impaired in HAZR group lacking Ad spermatogonia (stem cells for spermatozoa) indicating impaired mini puberty.

2010-11-23 | E-GEOD-25518 | biostudies-arrayexpress

Biomarkers in semen/urine from non-obstructive azoospermia patients to predict the presence of testicular spermatozoa and spermatogonia

Project description:About half of testicular sperm extraction (TESE) procedures in men with non-obstructive azoospermia (NOA), including men with Klinefelter syndrome (KS), are unsuccessful. To avoid unnecessary invasive surgery, biomarkers for spermatozoa have been studied and identified. In addition, markers for spermatogonia in testis tissue have been researched. This study aimed to find biomarkers in semen and/or urine of NOA patients to predict the presence of spermatogonia in the testis. This would especially be interesting for young boys. Differentially expressed miRNAs were identified (1) between samples from patients with and those of patients without a positive TESE procedure as well as (2) between TESE negative patients with and those without spermatogonia. A total of ten up-regulated miRNAs (seven in seminal plasma and three in urine) were found in the TESE-negative/spermatogonia-positive group compared to the TESE-negative/spermatogonia-negative group. These miRNAs could become biomarkers for spermatogonia, however, more research is necessary.

2023-02-06 | GSE224511 | GEO

Screening of genes involved in chromosome segregation in meiosis I

Project description:Prophase I of male meiosis involves dynamic chromosome segregation processes during early spermatogenesis, including synapsis, meiotic recombination, and cohesion. Genetic defects in genes participating in these processes consistently cause reproduction failure in mice. To identify candidate genes responsible for infertility in humans, we performed expression profiling of mouse spermatogenic cells undergoing meiotic prophase I. Cell fractions enriched in spermatogonia, leptotene/zygotene spermatocytes, or pachytene spermatocytes were separately isolated from mouse testes for RNA extraction. To minimize the contamination of other cell types, we fractionated the testicular cells undergoing the first round of spermatogenesis using Percoll gradient procedure. The cell fractions were characterized by morphological analysis by phase contrast microscopy and Nomarski interference microscopy, and then expression of cell lineage- and spermatogenesis stage-specific genes were examined by RT-PCR. The most enriched fractions for spermatogonia (fraction2 from day8), leptotene/zygotene spermatocytes (fraction5 from day12), and pachytene spermatocytes (fraction8 from day15) were subjected to hybridization on Affymetrix microarrays.

2010-03-31 | E-GEOD-19532 | biostudies-arrayexpress

Identification of type A spermatogonia specific transcriptome in rainbow trout immature testis

Project description:To identify the transcripts preferentially expressed in type A spermatogonia in rainbow trout immature testis, we compared the transcriptome between type A spermatogonia and testicular somatic cells by microarry analysis. We used fluorescence-activated cell sorting (FACS) to isolate type A spermatogonia or testicular somatic cells from the fishes carrying the transgene, pvasa-GFP, which expresses GFP in spermatogonia. RNA from type A spermatogonia and from testicular somatic cells were hybridized to a microarray after Cy3 labeling.

2012-06-11 | E-GEOD-38647 | biostudies-arrayexpress

Dissecting the dynamic cellular transcriptional atlas of adult teleost testis development throughout the annual reproductive cycle

Project description:The successful proliferation and differentiation of testicular cells are vital for the survival, continuity and preservation of species. However, a comprehensive molecular orchestration underlying the spermatogenic process in teleost testis development throughout the annual cycle remains elusive. In this study, the testicular cells derived from adult black rockfish at distinct stages—Regressed, Regenerating, and Differentiating were dissected via single cell transcriptome sequencing. A continuous developmental trajectory of spermatogenic cells, from spermatogonia to spermatids, was delineated, elucidating the molecular events involved in the spermatogonia development. Subsequently, the dynamic regulation of gene expressions associated with spermatogonia proliferation and differentiation was observed across spermatogonia subgroups and various developmental stages. A bioenergetic transition from glycolysis to mitochondrial respiration of spermatogonia during annual developmental cycle was demonstrated, and a deeper level of heterogeneity and molecular characteristics was revealed by re-clustering analysis. The developmental trajectory of Sertoli cells in annual reproductive cycle was examined, and the divergence of Leydig cells and macrophage were explored. Additionally, the interaction network between testicular micro-environment somatic cells and spermatogenic cell were established. Overall, our study provides detailed information on both germ and somatic cells within teleost testis during annual reproductive cycle, along with insights into the molecular regulation of spermatogonia dynamics, which lay the foundation for spermatogenesis regulation and germplasm preservation of endangered species.

2024-02-26 | GSE252438 | GEO

Methylome study on human spermatogenic cells

Project description:DNA methylation is an important regulator of the chromatin structure and is necessary for gene expression stability and maintenance of cell identitiy. However, to which extent DNA methylation changes during the key differentiation steps of human spermatogenesis remains largely unkown. Here, we generated genome wide methylomes (Enzymatic Methyl-sequencing) of human undifferentiated spermatogonia (Undiff), differentiating spermatogonia (Diff), tetraploid spermatocytes (4C) and haploid spermatids (1C) obtained from testicular tissues of three patients with qualitatively and quantitatively normal spermatogenesis (control, CTR). We also generated Undiff, Diff and 4C methylomes from two patients with impaired spermatogenesis (cryptozoospermic, CZ). Our study is the first to report about DNA methylation changes during human spermatogenesis.

| EGAS00001007449 | EGA

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