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EGR4 is a master gene responsible for fertility in cryptorchidism

ABSTRACT: RNA was isolated from testicular biopsies of cryptorchid boys. Gene expression profiles for these samples were investigated using Affymetrix arrays

ORGANISM(S): Homo sapiens

PROVIDER: GSE16191 | GEO | 2009/09/01

SECONDARY ACCESSION(S): PRJNA117199

REPOSITORIES: GEO

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EGR4 is a master gene responsible for fertility in cryptorchidism

Project description:RNA was isolated from testicular biopsies of cryptorchid boys. Gene expression profiles for these samples were investigated using Affymetrix arrays Whole testicular tissue was gained from cryptorchid boys aged 10 Months to 4.5 years as well as from controls of identical age. Each biopsy sample was collected for RNA isolation and GeneChip hybridization and was stored in cold 1x phosphate buffered saline (PBS). The tissue was then filtered through 100-nm nylon gauze. Fractions were collected, washed three times in 1x PBS, and snap-frozen in liquid nitrogen prior to storage at -80Â°C. An aliquot of each fraction was fixed in 1x PBS containing 1% formaldehyde (Polio, France) and 1% fetal calf serum (Eurabbie, France) and analyzed by DNA flow cytometry to determine its relative DNA content. Cell pellets and tissues were sheared in RLT buffer (Qiagen) supplemented with 1% Î²-Mercaptoethanol and further processed using a Qiashredder (Qiagen). The suspension was centrifuged for 2 min at maximum speed and the clarified lysate was stored at -80Â°C. Total RNA isolation was performed using RNeasy Mini-Spin columns (Qiagen) according to standard protocols. RNA quality was monitored using RNA Nano 6000 Chips and the 2100 BioAnalyzer (Agilent). cRNA synthesis, human U133 plus 2.0 GeneChip hybridization.

2009-09-01 | E-GEOD-16191 | biostudies-arrayexpress

Differential Gene Expression in Cryptorchid Testes

Project description:<p>Differential Gene Expression in Cryptorchid Testes Study uses whole-genome RNA profiling of testicular biopsies to determine a potential causative role of isolated congenital cryptorchidism in azoospermia and/or infertility in the context of our previously published GeneChip data. Cryptorchid patients, aged 7 months to 5 years and otherwise healthy, were enrolled in this prospective study. During surgery, testicular tissue biopsies were obtained for histological examination and RNA-sequencing (RNA-seq). Fifteen patients were selected based on the histological results and were divided into two groups. Seven were classified as belonging to the high infertility risk (HIR) and eight to the low infertility risk (LIR) group. Cryptorchid boys in the high infertility risk group lacked transformation of gonocytes into Ad spermatogonia due to impaired mini puberty. This group of patients will be infertile despite successful surgery.</p>

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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

In Vitro Propagation of XXY Human Klinefelter Spermatogonia Stem Cells; a step towards new fertility opportunities

Project description:Human testicular cells of XY and XXY were isolated from cryopreserved tissue in an experimental testicular tissue banking program and propagated in long-term culture for 110 days.

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Effect of miR-106b~25 in mouse spermatogenesis and male fertility

Project description:We investigated the miRNA cluster miR-106b~25 as potential candidate causing male infertility. We analized single and double KO mice. Single KO show derregulation of multiple molecular pathways and disrupted early spermatogenesys, but retain fertility. Double KO show severely disrupted testicular histology and significantly reduced fertility.

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Microarray analysis of human spermatogenic dysfunction

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Effect of ectopic expression of sFRP1 on MDA-MB-231 human breast cancer cells in vitro and in vivo as xenograft

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Transcription profiling of human peripheral blood mononuclear cells after a brief bout of exercise alters gene expression PBMCs from early- and late-pubertal males

Project description:We compared PBMC genomic response to exercise in both early (EB) and late-pubertal boys (LB) Experiment Overall Design: Twenty healthy males (age range 8-18 y/o) participated in this study. A baseline blood sample was taken before the onset of exercise and immediately after 30-min exercise bout. PBMCs were isolated using OptiPrepï Density Gradient Medium (SIGMA). Total RNA was extracted using TRIzolÂ®. cRNA was hybridized onto Affymetrix U133+2 arrays (total of 40 chips).

2008-11-13 | E-GEOD-11761 | biostudies-arrayexpress

Intratumor Heterogeneity and Precision of Microarray-Based Predictors of Breast Cancer Biology and Clinical Outcome

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DNA methylation patterns in perpheral blood samples from pre- and post-pubertal boys and girls

Project description:The COPENHAGEN Puberty Study is a combined cross sectional and longitudinal population based cohort study of healthy Danish children and adolescents. The clinical evaluations were performed by trained physicians and included pubertal staging of breast development according to TannerÂ´s classification evaluated by palpation. As a measure of pubertal onset a testicular volume of 4 ml or a breast tanner stage of B2 or more was used for boys and girls, respectively. The mean age between two examinations where this threshold was reached was used as their age of pubertal onset. Pre- and post-pubertal samples from 20 girls and 31 boys, in total 102 samples, were normalised using a Subset quantile Within-Array Normalization (SWAN) procedure and probes containing SNPs in the CpG or extension sites were removed.

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