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Transcription profiling of rat testis cell cultures grown on Sertoli cell and fibroblast feeder cell cultures

ABSTRACT: Rat germ cells

ORGANISM(S): Rattus norvegicus

SUBMITTER: Nikolaus Schultz

PROVIDER: E-GEOD-830 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Defining the spermatogonial stem cell.

Hamra F Kent FK Schultz Nikolaus N Chapman Karen M KM Grellhesl Dana M DM Cronkhite Jennifer T JT Hammer Robert E RE Garbers David L DL

Developmental biology 20040501 2

Through the use of donor cells from transgenic rats expressing GFP exclusively in the germline, we have defined culture conditions where male germ cells lose (on STO cells) or maintain (on MSC-1 cells) stem cell activity. A cadre of germ cell transcripts strikingly decrease in relative abundance as a function of testis age or culture time on STO cells, but only a subset of these transcripts (approximately 248) remain elevated when cultured on MSC-1 cells. If specific gene expression regulates st ...[more]

PMID: 15110708

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Characterization of rodent Sertoli cell primary cultures.

Project description:Sertoli cells play a vital role in spermatogenesis by offering physical and nutritional support to the differentiating male germ cells. They form the blood-testis barrier and secrete growth factors essential for germ cell differentiation. Sertoli cell primary cultures are critical for understanding the regulation of spermatogenesis; however, obtaining pure cultures has been a challenge. Rodent Sertoli cell isolation protocols do not rule out contamination by the interstitial or connective tissue cells. Sertoli cell-specific markers could be helpful, but there is no consensus. Vimentin, the most commonly used marker, is not specific for Sertoli cells since its expression has been reported in peritubular myoid cells, mesenchymal stem cells, fibroblasts, macrophages, and endothelial cells, which contaminate Sertoli cell preparations. Markers based on transcription and growth factors also have limitations. Thus, the impediment to obtaining pure Sertoli cell cultures pertains to both the method of isolation and marker usage. The aim of this review is to discuss improvements to current methods of rodent Sertoli cell primary cultures, assess the properties of prepubertal versus mature Sertoli cell cultures, and propose steps to improve cellular characterization. Potential benefits of using contemporary approaches, including lineage tracing, specific cell ablation, and RNA-seq for obtaining Sertoli-specific transcript markers are discussed. Evaluating the specificity and applicability of these markers at the protein level to characterize Sertoli cells in culture would be critical. This review is expected to positively impact future work using primary cultures of rodent Sertoli cells.

| S-EPMC7685524 | biostudies-literature

Alterations in hepatic gene expression induced by chromic exposure to high levels of the SV40 LT/st oncoproteins (4P)

Project description:Hepatic gene expression induced by chromic exposure to high levels of the SV40 LT/st oncoproteins (4P) Affymetrix mouse genome U74Av2 platform was used, duplicate measurements for wild-type and transgenic animals

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Alterations in hepatic gene expression induced by chronic exposure to low levels of the SV40 LT/st oncoproteins (4C)

Project description:Hepatic gene expression induced by chromic exposure to low levels of the SV40 LT/st oncoproteins (4P) Affymetrix mouse genome U74Av2 platform was used, duplicate measurements for wild-type and transgenic animals

2014-12-22 | E-GEOD-6232 | biostudies-arrayexpress

Transcription profiling of porcine olfactory bulb neuroblasts and PK15 non-neuronal, epithelial origin cell cultures

Project description:The phenotypically characterized hTERT immortalized porcine olfactory bulb neuroblast cell line (OBGF400) was subjected to an extensive whole genome-scaled expression profile for establishing their use as an in vitro neuronal disease model system. Microarrays were used to provide a comprehensive knowledge underlying the genomic complexity and overall gene expression capacity of the immortalized OBGF400 cells. The analysis revealed the elaborate signaling mechanisms of this unique subpopulation of porcine neuronally committed progenitors that mirrors the intricate organization of postnatal neurongenic zones. SUBMITTER_CITATION: Transcriptome Profile and Cytogenetic Analysis of Immortalized Neuronally Restricted Progenitor Cells Derived from the Porcine Olfactory Bulb. Animal biotechnology 2009 vol:20 iss:4 page:186-215 Experiment Overall Design: Total cellular RNA extracts from independent OBGF400 (neuroblasts) and PK15 (non-neuronal, epithelial origin) cell cultures (9 each) were pooled into a total of 3 biological replicates per cell line. The concentrations and purity of the pooled RNA preparations were determined using the BioAgilent RNA assay prior to hybridization on Affymetrix GeneChipÂ® Porcine Genome Expression Arrays. To ascertain the genes that were preferentially expressed by the OBGF400 neuroblasts, we used the PK15 cellular array in an effort to exclude somatic cell background.

2009-11-10 | E-GEOD-16855 | biostudies-arrayexpress

The effect of trypsin-2 on human tongue squamous cell carcinoma cell line gene expression

Project description:The level of trypsin-2 has been shown to correlate with the malignancy and metastatic potential of many cancer. We used microarrays to detail how tryspin-2 overexpression changes the gene profile of human tongue squamous cell carcinoma cell line (HSC-3). We compared trypsin-2 overexpressing HSC-3 line to HSC-3 which was transfected with control vector and because of that did not expressed trypsin-2 at all.

2012-10-02 | E-GEOD-32056 | biostudies-arrayexpress

DDB1 Regulates Sertoli Cell Proliferation and Testis Cord Remodeling by TGF? Pathway.

Project description:Testis cords are the embryonic precursors of the seminiferous tubules. Development of testis cords is a key event during embryonic testicular morphogenesis and is regulated by multiple signaling molecules produced by Sertoli cells. However, the exact nature and the cascade of molecular events underlying testis cord development remain to be uncovered. In the current study, we explored the role of DNA damage binding protein 1 (DDB1) in Sertoli cells during mouse testis cord development. The genetic ablation of Ddb1 specifically in Sertoli cells resulted in the compromised Sertoli cell proliferation and disruption of testis cord remodeling in neonatal mice. This testicular dysgenesis persisted through adulthood, resulting in smaller testis and low sperm production. Mechanistically, we observed that the DDB1 degradation can stabilize SET domain-containing lysine methyltransferase 8 (SET8), which subsequently decreases the phosphorylation of SMAD2, an essential intracellular component of transforming growth factor beta (TGF?) signaling. Taken together, our results suggest an essential role of Ddb1 in Sertoli cell proliferation and normal remodeling of testis cords via TGF? pathway. To our knowledge, this is the first upstream regulators of TGF? pathway in Sertoli cells, and therefore it furthers our understanding of testis cord development.

| S-EPMC6947845 | biostudies-literature

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2014-10-27 | E-MTAB-3032 | biostudies-arrayexpress

Sertoli Cell Wt1 Regulates Peritubular Myoid Cell and Fetal Leydig Cell Differentiation during Fetal Testis Development.

Project description:Sertoli cells play a significant role in regulating fetal testis compartmentalization to generate testis cords and interstitium during development. The Sertoli cell Wilms' tumor 1 (Wt1) gene, which encodes ~24 zinc finger-containing transcription factors, is known to play a crucial role in fetal testis cord assembly and maintenance. However, whether Wt1 regulates fetal testis compartmentalization by modulating the development of peritubular myoid cells (PMCs) and/or fetal Leydig cells (FLCs) remains unknown. Using a Wt1-/flox; Amh-Cre mouse model by deleting Wt1 in Sertoli cells (Wt1SC-cKO) at embryonic day 14.5 (E14.5), Wt1 was found to regulate PMC and FLC development. Wt1 deletion in fetal testis Sertoli cells caused aberrant differentiation and proliferation of PMCs, FLCs and interstitial progenitor cells from embryo to newborn, leading to abnormal fetal testis interstitial development. Specifically, the expression of PMC marker genes ?-Sma, Myh11 and Des, and interstitial progenitor cell marker gene Vcam1 were down-regulated, whereas FLC marker genes StAR, Cyp11a1, Cyp17a1 and Hsd3b1 were up-regulated, in neonatal Wt1SC-cKO testes. The ratio of PMC:FLC were also reduced in Wt1SC-cKO testes, concomitant with a down-regulation of Notch signaling molecules Jag 1, Notch 2, Notch 3, and Hes1 in neonatal Wt1SC-cKO testes, illustrating changes in the differentiation status of FLC from their interstitial progenitor cells during fetal testis development. In summary, Wt1 regulates the development of FLC and interstitial progenitor cell lineages through Notch signaling, and it also plays a role in PMC development. Collectively, these effects confer fetal testis compartmentalization.

| S-EPMC5201236 | biostudies-literature

Identification of Sertoli cell-specific transcripts in the mouse testis and the role of FSH and androgen in the control of Sertoli cell activity.

Project description:The Sertoli cells act to induce testis differentiation and subsequent development in fetal and post-natal life which makes them key to an understanding of testis biology. As a major step towards characterisation of factors involved in Sertoli cell function we have identified Sertoli cell-specific transcripts in the mouse testis and have used the data to identify Sertoli cell-specific transcripts altered in mice lacking follicle-stimulating hormone receptors (FSHRKO) and/or androgen receptors (AR) in the Sertoli cells (SCARKO).Adult iDTR mice were injected with busulfan to ablate the germ cells and 50 days later they were treated with diphtheria toxin (DTX) to ablate the Sertoli cells. RNAseq carried out on testes from control, busulfan-treated and busulfan + DTX-treated mice identified 701 Sertoli-specific transcripts and 4302 germ cell-specific transcripts. This data was mapped against results from microarrays using testicular mRNA from 20 day-old FSHRKO, SCARKO and FSHRKO.SCARKO mice. Results show that of the 534 Sertoli cell-specific transcripts present on the gene chips, 85% were altered in the FSHRKO mice and 94% in the SCARKO mice (mostly reduced in both cases). In the FSHRKO.SCARKO mice additive or synergistic effects were seen for most transcripts. Age-dependent studies on a selected number of Sertoli cell-specific transcripts, showed that the marked effects in the FSHRKO at 20 days had largely disappeared by adulthood although synergistic effects of FSHR and AR knockout were seen.These studies have identified the Sertoli cell-specific transcriptome in the mouse testis and have shown that most genes in the transcriptome are FSH- and androgen-dependent at puberty although the importance of FSH diminishes towards adulthood.

| S-EPMC5731206 | biostudies-literature

Sertoli cells maintain Leydig cell number and peritubular myoid cell activity in the adult mouse testis.

Project description:The Sertoli cells are critical regulators of testis differentiation and development. In the adult, however, their known function is restricted largely to maintenance of spermatogenesis. To determine whether the Sertoli cells regulate other aspects of adult testis biology we have used a novel transgenic mouse model in which Amh-Cre induces expression of the receptor for Diphtheria toxin (iDTR) specifically within Sertoli cells. This causes controlled, cell-specific and acute ablation of the Sertoli cell population in the adult animal following Diphtheria toxin injection. Results show that Sertoli cell ablation leads to rapid loss of all germ cell populations. In addition, adult Leydig cell numbers decline by 75% with the remaining cells concentrated around the rete and in the sub-capsular region. In the absence of Sertoli cells, peritubular myoid cell activity is reduced but the cells retain an ability to exclude immune cells from the seminiferous tubules. These data demonstrate that, in addition to support of spermatogenesis, Sertoli cells are required in the adult testis both for retention of the normal adult Leydig cell population and for support of normal peritubular myoid cell function. This has implications for our understanding of male reproductive disorders and wider androgen-related conditions affecting male health.

| S-EPMC4140823 | biostudies-literature

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