MiRNA expression profiling of enriched male germ cell populations
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE36566 | GEO | 2012/03/17
SECONDARY ACCESSION(S): PRJNA153721
REPOSITORIES: GEO
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Project description:MicroRNA is essential for the process of spermatogonesis, however analysis of its change in expression within germ cells during this process has been limited. We set out to examine the change in the miRNA expression profile of highly enriched mouse germ cell populations in vairous developmental stages.These inlcuded postnatal gonocytes, day 7-9 spermatogonia as well as pachytene spermatocytes and round spermatids from adult mice Gonocytes (day1) and spermatogonia (day 7-9), pachytene spermatocytes and round spermadits (adult) were enrichged by 2-4% BSA gradient sedementation. Three biological replicates of each cell type were included.
2012-03-17 | E-GEOD-36566 | biostudies-arrayexpress
Project description:Although numerous miRNAs have been identified in the testis, their roles in regulating the highly specific events that occur in the different germ cell types throughout spermatogenesis remain largely unknown. Furthermore, whether male germ cell miRNA expression is altered in response to or as a consequence of exposure to a toxic agent is unknown. Here we examine miRNA expression profiles in pachytene spermatocytes and round spermatids obtained from control rats and from rats treated with a chronic low dose of cyclophosphamide, a male germ cell toxicant. We observed that pachytene spermatocytes and round spermatids display vastly different miRNA expression profiles, reflecting their different developmental stages and possibly influencing the cellular response to toxic insult. Chronic low dose cyclophosphamide treatment altered the miRNA profiles in both pachytene spermatocytes and round spermatids. Target prediction analyses revealed that miRNAs altered by cyclophosphamide treatment may be involved in the response to cellular stress and damage. However, many are also involved in processes that are crucial for proper germ cell development. This study suggests that pachytene spermatocytes and round spermatids display distinct miRNA profiles that can be altered by cyclophosphamide treatment. The observed changes may be part of a response and repair mechanism to cyclophosphamide-induced damage or a dysregulation that disrupts normal germ cell development.
2018-09-05 | GSE119473 | GEO
Project description:The proteome changes were quantified in RibosomeRPL39L-/- spermatocytes and elongated spermatids using TMT 6-plex, and in spermatogonia, spermatocytes, round spermatids and elongated spermatids using TMT 10-plex by LC-MS/MS.
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Project description:Microarray analysis of purified pachytene spermatocytes and round spermatids. Each stage was examined in wild type and RNF8 knockout mice in two biological replicates. We performed microarray analysis using Affymetrix Gene 1.0 ST Arrays with purified pachytene spermatocytes and round spermatids. Pachytene spermatocytes and round spermatids were enriched from 3 to 4 males from the WT or Rnf8-KO via BSA gravity sedimentation according to the previous publication [PMID 8231890] and >95% (PS, RS) enrichments were verified after DAPI staining under a fluorescent microscope. For microarray analysis, total RNAs from purified pachytene spermatocytes or round spermatids were examined.
2013-01-04 | E-GEOD-39302 | biostudies-arrayexpress
Project description:In order to monitor the changes in small RNAs expression during mouse spermatogenesis. Type A spermatogonia, pachytene spermatocytes and round spermatids were isolated following collagenase treatment of testes and trypsin digestion of isolated seminiferous tubules using unit gravity sedimentation in a bovine serum albumin gradient. The small RNA fraction (18-36nt) was cloned and sequenced from total RNA of each cell type. Three types of mouse germ cells small RNAs were used for high throughput sequencing analysis
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Project description:Mouse Round Spermatids (Sptd) and Pachytene Spermatocytes (Spcy) Keywords: repeat sample
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Project description:Mammalian spermatogenesis is a complex biological process that occurs within a highly organized tissue, the seminiferous epithelium. The coordinated maturation of spermatogonia, spermatocytes and spermatids suggests the existence of precise programs of gene expression in these cells as well as in their neighboring somatic Sertoli cells. The objective of this study was to elucidate genes encoding the proteins that execute these programs. Rat seminiferous tubules at stages I, II-III, IV-V, VI, VIIa,b, VIIc,d, VIII, IX-XI, XII, XIII-XIV of the cycle were isolated by microdissection and Sertoli cells, spermatogonia plus early spermatocytes, pachytene spermatocytes and spermatids were purified from enzymatically-dispersed testes. Microarray analysis using Rat Genome 230 2.0 arrays identified a total of 16,971 probe sets that recognized transcripts. A comparison with the transcriptome of other tissues identified 398 testis-specific probe sets, which therefore are potential targets for the development of new contraceptives. Sequential waves of cell and stage-specific gene expression are associated with progression of germ cells through the stages of the cycle of the seminiferous epithelium and 1612 probe sets recognized transcripts whose expressions varied at least 4-fold across the stages of the cycle. Pathway analyses reveal that entire biological processes are regulated cyclically in testicular cells. Important among these are cell cycle and DNA repair. Thus, stage-specific gene expression is a widespread and fundamental characteristic of spermatogenic cells and Sertoli cells. Experiment Overall Design: Seminiferous tubules at the following stages or groups of stages were isolated by transillumination-assisted microdissection: I, II-III, IV-V, VI, VIIa,b, VIIc,d, VIII, IX-XI, XII, XIII-XIV. Twenty-five segments of tubules, 2 m in length, were collected at each stage or group of stages from each rat. Tubules from two separate animals were pooled to form one sample from each stage or groups of stages. A total of 5 independent samples per stage or groups of stages were analyzed. Mature Sertoli cells were isolated to about 85% purity (n=4). Three groups of germ cells were isolated by centrifugal elutriation from enzyme-dispersed testes: round spermatids (n=4), pachytene spermatocytes (n=4) and spermatogonia plus early spermatocytes (n=3) . Purity of these cells has been shown to vary between 85% to 95%
2008-06-20 | E-GEOD-8978 | biostudies-arrayexpress
Project description:To further understand the effects of chronic cyclophosphamide treatment on spermatogenesis, we used whole genome microarrays to identify differentially expressed genes in pachytene spermatocytes and round spermatids from treated and control male rats. Pachytene spermatocytes and round spermatids from rats treated chronically with cyclophosphamide were isolated and profiled for changes in gene expression.
2016-03-23 | E-GEOD-79471 | biostudies-arrayexpress
Project description:To determine the dynamics of open chromatin at a genomic resolution during spermatogenesis, we performed ATAC-seq and detected genomic regions of accessible chromatin by Tn5 transposase during spermatogenesis. We analyzed four representative stages of spermatogenesis: Thy1+ undifferentiated spermatogonia, which contains spermatogonial stem cells and progenitor cells; c-Kit+ differentiating spermatogonia from P7 testes; purified pachytene spermatocytes (PS) undergoing meiosis; and postmeiotic round spermatids (RS) from adult testes
2017-10-18 | GSE102954 | GEO