Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression data of pachytene spermatocytes and round spermatids from young and aged Brown Norway rats

ABSTRACT: Male fertility and testis function changes with age and so it was sought to determine if these changes are accompanied by changes in gene expression. A full genome microarray was used to determine if distinct pathways of genes were altered in expression in germ cells (pachytene spermatocytes or round spermatids) with age. PACH: Testes from young (4 months) and aged (18 months) Brown Norway rats were subjected to a density gradient cell separation to isolate pachytene spermatocytes. RNA was isolated from these cells for hybridization on affymetrix miroarrays. ROU: Testes from young (4 months) and aged (18 months) Brown Norway rats were subjected to a density gradient cell separation to isolate round spermatids. RNA was isolated from these cells for hybridization on affymetrix miroarrays.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Bernard Robaire

PROVIDER: E-GEOD-29963 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Aging results in differential regulation of DNA repair pathways in pachytene spermatocytes in the Brown Norway rat.

Paul Catriona C Nagano Makoto M Robaire Bernard B

Biology of reproduction 20110824 6

The present trend of increasing paternal age is accompanied by concerns for the development of complex multigene diseases (e.g., autism and schizophrenia) in progeny. Recent studies have established strong correlations between male age, increased oxidative stress, decreased sperm quality, and structural aberrations of chromatin and DNA in spermatozoa. We tested the hypothesis that increasing age would result in altered gene expression relating to oxidative stress and DNA damage/repair in germ ce ...[more]

PMID: 21865553

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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%

Dataset Information

Expression data of pachytene spermatocytes and round spermatids from young and aged Brown Norway rats

Publications

Aging results in differential regulation of DNA repair pathways in pachytene spermatocytes in the Brown Norway rat.

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