ABSTRACT: miRNA expression profiling of isolated populations of prepachytene spermatocytes, pachytene spermatocytes and spermatids of PWD and B6 male mice
Project description:MiRNA expression profiling of isolated populations of prepachytene spermatocytes (LP), pachytene spermatocytes (RP) and spermatids (ST) from PWD and B6 was performed to study the genome wide variation in miRNA expression between two mouse subspecies. Populations of pre-pachytene spermatocytes, pachytene spermatocytes and spermatids were isolated from PWD and B6 mice. All populations were isolated in triplicates from two bilogical replicas Hence, RNA was prepared from 18 samples and was then subject to labeling and hybridization on microarray chips.
Project description:Expression profiling of isolated populations of prepachytene spermatocytes (LP), pachytene spermatocytes (RP) and spermatids (ST) from PWD and B6 was performed to study the genome wide variation in gene expression between two mouse subspecies. To evaluate the transcriptional difference between B6 and PWD in during meiosis, we compared their transcriptomes in sorted populations of pre-pachytene primary spermatocytes (Leptonema, Zygotene and Pachytene), pachytene spermatocytes (Mid-late pachytene and diplotene) and spermatids. Populations of pre-pachytene spermatocytes, pachytene spermatocytes and spermatids were isolated from PWD and B6 mice. All populations were isolated in duplicates from two animals. Hence, RNA was prepared from 12 samples (= 2 strains x 3 tissues x 2 replicas) and was then subject to labeling and hybridization on microarray chips.
Project description:Expression profiling of isolated populations of prepachytene spermatocytes (LP), pachytene spermatocytes (RP) and spermatids (ST) from PWD and B6 was performed to study the genome wide variation in gene expression between two mouse subspecies. To evaluate the transcriptional difference between B6 and PWD in during meiosis, we compared their transcriptomes in sorted populations of pre-pachytene primary spermatocytes (Leptonema, Zygotene and Pachytene), pachytene spermatocytes (Mid-late pachytene and diplotene) and spermatids.
Project description:MiRNA expression profiling of isolated populations of prepachytene spermatocytes (LP), pachytene spermatocytes (RP) and spermatids (ST) from PWD and B6 was performed to study the genome wide variation in miRNA expression between two mouse subspecies.
Project description:Expression profiling of isolated populations of prepachytene spermatocytes, pachytene spermatocytes and spermatids of PWD and B6 males
Project description:Expression profiling of isolated populations of prepachytene spermatocytes, pachytene spermatocytes and spermatids of males carrying a translocation T(16;17)43H was performed to study the influence of this translocation on gene expression from individual chromosomes. To evaluate the transcriptional behavior of individual chromosomes in the sterile B10-T43/+ males and their fertile congenic B10-T43/T43 and B10 counterparts, we compared their transcriptomes in sorted populations of pre-pachytene primary spermatocytes, pachytene primary spermatocytes and spermatids. Keywords: individual genetic characteristics design
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.
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.
Project description:Comparison of gene expression differences between Dnmt3L heterozygous and wildtype pachytene spermatocytes, and similarly between Dnmt3L heterozygous and wildtype round spermatids which were isolated from the Dnmt3L knockout mouse line. This array was conducted to address the hypothesis that Dnmt3L heterozygosity results in deregulated gene expression within spermatocytes and spermatids. Results show that Dnmt3L heterozygosity causes numerous genes to be differentially regulated on a genome-wide level, showing that DNMT3L has an important role in regulating gene expression within these male germ cells. Three preparations each of Dnmt3L purified wildtype spermatocytes, wildtype spermatids, heterozygous spermatids, and two preparations of heterozygous spermatocytes were isolated for a total of 11 samples. Each preparation was made up of cells isolated from 10 mice.
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.
