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Microarrays analysis of RNA isolated from 7days-Sertoli cell culture of 14 day-old AMH-Cre/WT and WT/WT C57BL/6J mouse testes

ABSTRACT: In order to understand whether sole Cre recombinase expression has an effect on stress signalling pathways and the peroxisomal or other subcellular compartments, we have analyzed total RNA isolated from Sertoli cell cultures of anti-Müllerian-hormone (AMH)-Cre/wild-type and wild-type/wild-type C57BL/6J mouse testes using microarrays. The Sertoli cells were isolated from individual 14 day-old mice and were cultivated for 7 days. Total RNA was isolated from the Sertoli cell culture (for AMH-Cre/WT 5 mice and WT/WT 3 mice) and the same genotypes were pooled together. For the microarray analysis two technical replicates for each genotype were generated.

ORGANISM(S): Mus musculus

SUBMITTER: Eveline Baumgart-Vogt

PROVIDER: E-GEOD-28307 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Microarrays analysis of RNA isolated from 7days-Sertoli cell culture of 14 day-old AMH-Cre/WT and WT/WT C57BL/6J mouse testes

Project description:In order to understand whether sole Cre recombinase expression has an effect on stress signalling pathways and the peroxisomal or other subcellular compartments, we have analyzed total RNA isolated from Sertoli cell cultures of anti-Müllerian-hormone (AMH)-Cre/wild-type and wild-type/wild-type C57BL/6J mouse testes using microarrays.

2012-01-01 | GSE28307 | GEO

Specific deletion of protein phosphatase 6 catalytic subunit in Sertoli cells leads to disruption of spermatogenesis

Project description:Protein phosphatase 6 (PP6) is a member of the PP2A-like subfamily, which plays significant roles in numerous fundamental biological activities. We found that PPP6C plays important roles in male germ cells recently. Spermatogenesis is supported by the Sertoli cells in seminiferous epithelium. In this study, we crossed Ppp6cF/F mice with AMH-Cre mice to gain mutant mice with specific depletion of the Ppp6c gene in the Sertoli cells. We discovered that the PPP6C cKO male mice were absolutely infertile and germ cells were largely lost during spermatogenesis. By combing phosphoproteome with bioinformatics analysis, we showed that the phosphorylation status of β-catenin at S552 (a marker of adherens junctions) was significantly upregulated in mutant mice. Abnormal β-catenin accumulation resulted in impaired testicular junction integrity, thus led to abnormal structure and functions of BTB. Taken together, our study reveals a novel function for PPP6C in male germ cell survival and differentiation by regulating the cell-cell communication through dephosphorylating β-catenin at S552.

2022-02-15 | PXD028051 | Pride

Constitutive expression of activated NOTCH1 in fetal Sertoli cells dysregulates factors required for proper gonocyte maintenance and survival.

Project description:Genome-wide analysis of whole embryonic day 14.5 mouse testes constitutively expressing the activated intracellular domain of NOTCH1 in Sertoli cells. The goal of the study was to identify Sertoli cell expressed genes regulated by Notch signaling, which are critical to gonocyte maintenance and survival. The mouse model was generated by intercrossing anti-Müllerian hormone (Amh)-cre transgenic mice and RosaNICD transgenic mice. Testes were dissected free from mesonephroi and testis pairs from a single fetus were pooled to generate a single sample. Littermates not expressing Amh-cre were used as controls. Testes from one control and one overexpressor fetus per dam from three separate dams were used for analysis.

2013-02-08 | E-GEOD-37073 | biostudies-arrayexpress

Constitutive expression of activated NOTCH1 in fetal Sertoli cells dysregulates factors required for proper gonocyte maintenance and survival.

2013-02-08 | GSE37073 | GEO

Seminiferous tubule degeneration and infertility in mice with sustained activation of WNT/CTNNB1 signaling in sertoli cells

Project description:To elucidate the role(s) of WNT/CTNNB1 signaling in the testis, transgenic Ctnnb1 tm1Mmt/+;Amhr2 tm3(cre)Bhr/+ mice were generated to obtain sustained activation of the WNT/CTNNB1 pathway in Sertoli cells. Male Ctnnb1 tm1Mmt/+;Amhr2 tm3(cre)Bhr/+ mice were sterile because of testicular atrophy starting at 5 wk of age, associated with degeneration of seminiferous tubules and the progressive loss of germ cells. Sustained WNT/CTNNB1 pathway activation was obtained in Ctnnb1 tm1Mmt/+;Amhr2 tm3(cre)Bhr/+ Sertoli cells. Sertoli cells often exhibited morphological characteristics suggestive of incomplete differentiation that appeared in a manner coincident with germ cell loss, accompanied by an increase in the expression of the immature Sertoli cell marker AMH. Microarray analyses performed on cultured Sertoli cells showed that CTNNB1 induces the expression of genes associated with the female sex determination pathway as well as cell-cycle associated genes . Together, these data suggest that the WNT/CTNNB1 pathway regulates Sertoli cell functions critical to their capacity to support spermatogenesis and to proliferate in the postnatal testis. Sertoli cells were obtained from 3wks-old Ctnnb1(tm1Mmt/tm1Mmt) mice. Cells were infected with adenoviruses to express eGFP or Cre in serum-free medium, and subsequently harvested for RNA extraction. Preliminary experiments demonstrated that an infection efficiency of nearly 100% could be obtained at an MOI of 50 (as determined by analysis of fluorescent signal in Ad-eGFP-infected cells) and that the Ad-Cre virus produced complete recombination of the floxed Ctnnb1 allele after 12 hours. Microaaray analyses were done using triplicate RNA samples from each adenoviral treatment using MouseRef-6 v.2.0 expression BeadChips technology (Illumina, San Diego, CA).

2011-04-05 | E-GEOD-28402 | biostudies-arrayexpress

IGR_U693_AR_GE_STUDY

Project description:The androgen receptor is a steroid receptor belonging to the superfamily of hormone-activated transcriptional factors, displaying distinct expression profiles in Sertoli cells during testis development, tightly correlated to the stages of spermatogenesis.The aim of the project was to better understand the AR signaling pathways, and identify androgen regulated genes in a mature Sertoli cell line (ST38c). In order to identify androgen regulated candidate genes we compared the gene expression of mature Sertoli cells (ST38c) in the absence (condition B) and in the presence of a androgenic ligand, dihydrotestosterone (condition ST). In order to understand the AMH gene repression in mature Sertoli cells, which occurs at puberty, soon after the augmentation of the AR expression, we also compared the gene profile of an immature, not expressing the androgen receptor, prepubertal Sertoli cell line (SMAT1-condition A) with the mature Sertoli cell line (ST38c- condition C), expressing the androgen receptor. The Sertoli cell line SMAT1 has been obtaiend from a 6-day old transgenic male mouse (using the targeted oncogenesis SV40). This cell line has been kindly provided to us by Dr Jean Yves Picard (UMR 782), Universite Paris Sud. (Dutertre M, et al., Mol Cell Endocrinol. 1997 Dec 31;136(1):57-65. PMID 9510068).The Sertoli cell line ST38c was obtained in the INSERM U-693 from 8-wk-old male murine testis; from a transgenic mouse carrying the construct in which the SV40 large T Antigen (TAg) was placed under the control of the human vimentin promoter. The development and description of this new cell line have not yet been published.

2014-07-01 | E-MTAB-1732 | biostudies-arrayexpress

Effects of deletion of Arhgef15 gene in mouse sertoli cells on spermatogenesis

Project description:Sertoli cells, which are the only somatic cells in the seminiferous tubules, facilitate the maintenance of testicular immune privilege through the formation of the blood-testis barrier (BTB) and the expression of immunoregulatory factors. Rho guanosine exchange factor 15 (ARHGEF15) is a member of guanosine exchange factors, which are involved in cell migration, cell polarity, and cell cycle progression via activation of Rho GTPases. This study investigated the functions of ARHGEF15 in Sertoli cells during spermatogenesis using Sertoli cell–specific Arhgef15 knockout mice. The results showed that Arhgef15 deficiency in Sertoli cells affected the localization of Sertoli cell nuclei, disrupted BTB integrity, and led to premature shedding of germ cells. In Arhgef15flox/flox/Amh-Cre+ mice, the ultrastructure of the round spermatids was impaired, accompanied by acrosome degeneration, acrosomal vesicle shedding, and atrophic nuclei. Consequently, the percentage of abnormal sperm in the Arhgef15flox/flox/Amh-Cre+ epididymis was markedly elevated. RNA-sequencing analysis revealed that most of the differentially expressed genes in Sertoli cells of Arhgef15flox/flox/Amh-Cre+ mice were related to immunity. Further study revealed that the sera of Arhgef15flox/flox/Amh-Cre+ mice showed immunoreactivity against testicular lysate of wild-type mice, indicating the production of antibodies against testicular autoantigens in Arhgef15flox/flox/Amh-Cre+ mice. In conclusion, the specific deletion of Arhgef15 in Sertoli cells leads to sperm abnormality, probably by disrupting the testicular immune homeostasis, in mice.

2022-06-16 | GSE206018 | GEO

Transcription profiling of mice mutant for Sox8 and Sox9 has early testicular sterility

Project description:To study a possible redundant role of mouse Sox8 and Sox9 after testis induction we generated double mutants by AMH-Cre conditional inactivation of Sox9 in a Sox8-/- background. Double mutants arrest spermatogenesis in early pubescence. We performed expression profiling to explore causes. AMH-Cre/+;Sox9flox/flox;Sox8-/- is a conditional homozygous knockout of SOX9 in a sox8-/- background, using an AMH-CRE line where the CRE recombinase is expressed under the promoter region of the human AMH gene in Sertoli cells starting at E13.5. The specific cross was between males of the genotype +/+;Sox9flox/flox;Sox8-/- and females of the genotype AMH-Cre/+;Sox9flox/flox;Sox8+/-.

2013-01-03 | E-TABM-531 | biostudies-arrayexpress

Riding the spermatogenic wave: Profiling gene expression within neonatal germ and Sertoli cells during a synchronized initial wave of spermatogenesis

Project description:WIN 18,446/RA treatment of neonatal mice was used to synchronize the initial wave of spermatogenesis and identify novel messages expressed within either germ or Sertoli cells as spermatogonia enter meiosis. germ cell-specific (Stra8-cre: RiboTag; or Ngn3-cre:RiboTag) and Sertoli cell-specific (Amh-Cre: RiboTag)

2014-05-01 | E-GEOD-54408 | biostudies-arrayexpress

Sertoli cell specific testicular gene expression of Connexin 43 knockout mice

Project description:Infertility observed in adult Sertoli cell (SC)-specific Connexin 43 Knockout-mice rather seems to be an effect of the disturbed SC-Germ cell (GC) crosstalk than a direct consequence of the loss of Cx43 protein in SC with important GC specific genes being mostly affected by this deletion. Identification of differentially expressed genes in testis of cx43 KO-mice at developmental stage 8 days post partum when compared with testis of WT-mice

2011-01-06 | E-GEOD-23431 | biostudies-arrayexpress

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