Sertoli Cell Lines Established fromH-2Kb-tsA58 Transgenic Mice Differentially Regulate the Expression of Cell-Specific Genes

Walther, Norbert; Jansen, Martina; Ergün, Süleyman; Kascheike, Bernd; Ivell, Richard

doi:10.1006/excr.1996.0192

Cited by 56 publications

(43 citation statements)

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“…RNA was extracted from mouse testis and mouse-derived SK49 (Walther et al 1996) and TM4 (Mather 1980) Sertoli cell lines using the FastRNA Pro Green kit (Qbiogene, Illkirch Cedex, France), according to manufacturer's instructions. Two micrograms of each total RNA fraction were reverse transcribed in a 20 ml volume, using random hexamers and the Superscript II pre-amplification system (Invitrogen), according to manufacturer's instructions.…”

Section: Rt-pcrmentioning

confidence: 99%

Phosphoprotein enriched in astrocytes-15 is expressed in mouse testis and protects spermatocytes from apoptosis

Mizrak

Raffi²,

Párraga³

et al. 2007

Reproduction

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Phosphoprotein enriched in astrocytes (PEA-15) is a 15 kDa acidic serine-phosphorylated protein expressed in different cell types, especially in the CN. We initially detected the expression of PEA-15 in primary cultures of Sertoli cells. To assess the presence and localization of PEA-15 in the mouse testis, we studied the expression pattern of the PEA-15 protein by immunohistochemistry and mRNA by in situ hybridization. Both the protein and the mRNA of PEA-15 were localized in the cytoplasm of Sertoli cells, all types of spermatogonia, and spermatocytes up till zygotene phase of the meiotic prophase. Subsequently, with ongoing development of the spermatocytes, the expression decreased and was very low in the cytoplasm of diplotene spermatocytes. To analyze the possible role of PEA-15 in the developing testis, null mutants for PEA-15 were examined. As the PEA-15 C terminus contains residues for ERK binding, we studied possible differences between the localization of the ERK2 protein in wild type (WT) and PEA-15 K/K mice. In the WT testis, ERK2 was localized in the cytoplasm of Sertoli cells, B spermatogonia, preleptotene, leptotene, and zygotene spermatocytes, whereas in the KO testis, ERK2 was primarily localized in the nuclei of these cells and only little staining remained in the cytoplasm. Moreover, in PEA-15-deficient mice, significantly increased numbers of apoptotic spermatocytes were found, indicating an anti-apoptotic role of PEA-15 during the meiotic prophase. The increased numbers of apoptotic spermatocytes were not found at a specific step in the meiotic prophase.

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Section: Rt-pcrmentioning

confidence: 99%

Phosphoprotein enriched in astrocytes-15 is expressed in mouse testis and protects spermatocytes from apoptosis

Mizrak

Raffi²,

Párraga³

et al. 2007

Reproduction

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“…Upon transfer to the non-permissive temperature of 39.5C, the temperature-sensitive SV40 T-antigen is inactivated, leading to a more differentiated state of the cells characterised by the expression of many important markers of Sertoli cells and the cessation of growth (Walther et al 1996(Walther et al , 1997.…”

Section: Sertoli-cell-derived Cell Linesmentioning

confidence: 99%

Gene expression profiling of mouse Sertoli cell lines

Strothmann¹,

Simoni²,

Mathur

et al. 2004

Cell and Tissue Research

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The proliferation and differentiation of Sertoli cells is regulated by follicle-stimulating hormone (FSH). The molecular events following FSH stimulation are only partially known. To investigate FSH action in Sertoli cells, we established two novel FSH-responsive mouse Sertoli-cell-derived lines expressing human wild-type (WT) FSH receptor (FSHR) or overexpressing mutated (Asp567Gly) constitutively active FSHR (MUT). Gene expression profiling with commercially available cDNA arrays, including 588 mouse genes, revealed 146 genes expressed in both cell lines. Compared with the expression pattern of WT cells, 20 genes were identified as being either up- or down-regulated (>two-fold) in the MUT cells. We observed a strong differential expression of factors involved in cellular proliferation, e.g. cyclin D2 (repressed to nearly undetectable levels), proliferating cell nuclear antigen (2.5-fold repression) and Eps-8 (six-fold repression), and in genes involved in cellular differentiation, e.g. cytokeratin-18 (13-fold induction). The cDNA array results for six representative genes were confirmed by Northern blotting, which also included the parental SK-11 cell line devoid of FSHR expression. We found no further acute FSH- or forskolin-induced change in expression levels after 3-h stimulations, suggesting that the observed differences between the two cell lines is a consequence of mild, chronically increased, cAMP production in MUT cells. These results provide a platform for the further investigation of selected candidate genes in primary cultures and/or in vivo.

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“…Sertoli cell lines from adult mouse testis designed using published sequences from mouse or other mammalian species (Walther et al 1996, Schrans-Stassen et al 1999, Beverdam et al 2003, Hofmann et al 2003 or Primer BLAST (NCBI). For the RT-PCR of Fshr, after the first-round PCR, the amplified products (50 times diluted) were used for a second-round nested PCR.…”

Section: Detection Of Sertoli Cell Transcripts By Rt-pcr For Final CLmentioning

confidence: 99%

Establishment of adult mouse Sertoli cell lines by using the starvation method

Sato¹,

Yoshida²,

Nozawa³

et al. 2013

Reproduction

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Sertoli cells were isolated from the testes of 6-week-old mice and stable Sertoli cell lines with higher proliferation rates were subcloned after starvation of primary cultured cells. After two rounds of this subcloning, 33 subcloned lines were selected on the basis of their proliferation rates. In addition, these subclones were screened according to their phagocytic activity and the characteristics of mature Sertoli cells, such as the expression of androgen receptors (ARs) and progesterone receptors, by using western blotting and immunocytochemical analysis, in addition to their morphology and proliferation rates. After the third round of subcloning, 12 subclones were selected for the final selection using RT-PCR for identification of genes specifically expressed by various testicular cells. Three clones were selected that expressed Sertoli-cell-specific genes, i.e. stem cell factor, clusterin, AR, a-inhibin, transferrin, Wilms' tumour-1, Mü llerian inhibitory substance, sex-determining region Y-box 9, FSH receptor (Fshr) and occludin; however, these clones did not express globulin transcription factor 1, steroidogenic factor or androgen-binding protein. These clones also expressed growth and differentiation factors that act on germ cells, such as leukaemia inhibitory factor, transforming growth factor b1 and basic fibroblast growth factor 2, but did not express c-kit (specific for germ cells), LH receptor and 3b-hydroxyl-dehydrogenase (specific for Leydig cells). Immunocytochemical data confirmed the expression of clusterin in these clones. Furthermore, the Bromodeoxyuridine incorporation assay confirmed the proliferation activity of these clones through Fshr after treatment with FSH. These clones are considered to be valuable tools for the study of Sertoli cell-specific gene expression and function. Reproduction (2013) 145 505-516

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Sertoli Cell Lines Established fromH-2Kb-tsA58 Transgenic Mice Differentially Regulate the Expression of Cell-Specific Genes

Cited by 56 publications

References 0 publications

Phosphoprotein enriched in astrocytes-15 is expressed in mouse testis and protects spermatocytes from apoptosis

Phosphoprotein enriched in astrocytes-15 is expressed in mouse testis and protects spermatocytes from apoptosis

Gene expression profiling of mouse Sertoli cell lines

Establishment of adult mouse Sertoli cell lines by using the starvation method

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