Identification of Developmentally Regulated Genes in the Somatic Cells of the Mouse Testis Using Serial Analysis of Gene Expression1

O’Shaughnessy, Peter J.; Fleming, Lynne; Baker, Paul; Jackson, Gary L.; Johnston, Heather

doi:10.1095/biolreprod.103.016899

Cited by 44 publications

(29 citation statements)

References 68 publications

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“…Best1 transcripts were first evident at mouse postnatal day (PN) 19, with a strong increase until PN30, where it reached steady-state adulthood levels. This pattern is similar to that of the known sperm-specific Ca 2+ channel CatSper1 but not Leydig/Sertoli cell markers 17βHsdIII and Ptgds (27) (Fig. 1E).…”

Section: Expression Profiling Suggests a Role For Best1 In Mouse Spersupporting

confidence: 78%

Bestrophin 1 is indispensable for volume regulation in human retinal pigment epithelium cells

Milenkovic¹,

Brandl

Milenkovic³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

115

142

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In response to cell swelling, volume-regulated anion channels (VRACs) participate in a process known as regulatory volume decrease (RVD). Only recently, first insight into the molecular identity of mammalian VRACs was obtained by the discovery of the leucine-rich repeats containing 8A (LRRC8A) gene. Here, we show that bestrophin 1 (BEST1) but not LRRC8A is crucial for volume regulation in human retinal pigment epithelium (RPE) cells. Whole-cell patch-clamp recordings in RPE derived from human-induced pluripotent stem cells (hiPSC) exhibit an outwardly rectifying chloride current with characteristic functional properties of VRACs. This current is severely reduced in hiPSC-RPE cells derived from macular dystrophy patients with pathologic BEST1 mutations. Disruption of the orthologous mouse gene (Best1 −/− ) does not result in obvious retinal pathology but leads to a severe subfertility phenotype in agreement with minor endogenous expression of Best1 in murine RPE but highly abundant expression in mouse testis. Sperm from Best1 −/− mice showed reduced motility and abnormal sperm morphology, indicating an inability in RVD. Together, our data suggest that the molecular identity of VRACs is more complex-that is, instead of a single ubiquitous channel, VRACs could be formed by cell type-or tissue-specific subunit composition. Our findings provide the basis to further examine VRAC diversity in normal and diseased cell physiology, which is key to exploring novel therapeutic approaches in VRAC-associated pathologies.bestrophin 1 | volume-regulated anion channel | induced pluripotent stem cell | retinal pigment epithelium | mouse sperm

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Section: Expression Profiling Suggests a Role For Best1 In Mouse Spersupporting

confidence: 78%

Bestrophin 1 is indispensable for volume regulation in human retinal pigment epithelium cells

Milenkovic¹,

Brandl

Milenkovic³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

115

142

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“…AMH causes the Müllerian duct to degenerate in XY embryos and promotes testicle differentiation. It has been reported that Amh and Krt18 are expressed in immature testes, but are absent in adult testes [13,67,68]. Unexpectedly, we detected both of them in adult male mice.…”

Section: Differential Expression Of Biomarkers In Testisspecific Cellssupporting

confidence: 53%

“…The advent of serial analysis of gene expression (SAGE) has made these technologies more efficient and O'Shaughnessy et al have used SAGE to generate gene expression profiles from the somatic cells of fetal and adult mouse testes [13]. They identified 12976 unique transcripts and found that most of the Sertoli or Leydig cell-specific genes are up-regulated in adult mice.…”

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confidence: 99%

Transcriptome profiling of the developing postnatal mouse testis using next-generation sequencing

Gong

Pan

Lin

et al. 2012

Sci. China Life Sci.

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Mammalian testis development is a complex and highly sophisticated process. To study the dynamic change of normal testis development at the transcriptional level, we investigated mouse testes at three postnatal ages: 6 days postnatal, 4 weeks old, and 10 weeks old, representing infant (PN1), juvenile (PN2), and adult (PN3) stages, respectively. Using ultra high-throughput RNA sequencing (RNA-seq) technology, we obtained 211 million reads with a length of 35 bp. We identified 18837 genes that were expressed in mouse testes, and found that genes expressed at the highest level were involved in spermatogenesis. The gene expression pattern in PN1 was distinct from that in PN2 and PN3, which indicates that spermatogenesis has commenced in PN2. We analyzed a large number of genes related to spermatogenesis and somatic development of the testis, which play important roles at different developmental stages. We also found that the MAPK, Hedgehog, and Wnt signaling pathways were significantly involved at different developmental stages. These findings further our understanding of the molecular mechanisms that regulate testis development. Our study also demonstrates significant advantages of RNA-seq technology for studying transcriptome during development. next-generation sequencing, transcriptome, mouse testis, development Citation:Gong W, Pan L L, Lin Q, et al. Transcriptome profiling of the developing postnatal mouse testis using next-generation sequencing.

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“…Poly (A) C RNA was isolated from equal amounts of RNA pooled from four individual follicles at the COH, SF and DF stages using oligo dT cellulose (Invitrogen) or the Oligotex mRNA Mini Kit (Qiagen Ltd). The SAGE libraries were constructed from poly (A) C RNA as described previously (Velculescu et al 1995, O'Shaughnessy et al 2003, McRae et al 2005. The ditags were amplified in two different stages of 23 cycles and 10 or 11 cycles using nested biotinylated primers.…”

Section: Construction Of Sage Librariesmentioning

confidence: 99%

Differentiation of the bovine dominant follicle from the cohort upregulates mRNA expression for new tissue development genes

Mihm¹,

Baker²,

Fleming³

et al. 2008

Reproduction

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This study was designed to identify genes that regulate the transition from FSH-to LH-dependent development in the bovine dominant follicle (DF). Serial analysis of gene expression (SAGE) was used to compare the transcriptome of granulosa cells isolated from the most oestrogenic growing cohort follicle (COH), the newly selected DF and its largest subordinate follicle (SF) which is destined for atresia. Follicle diameter, follicular fluid oestradiol (E) and E:progesterone ratio confirmed follicle identity. Results show that there are 93 transcript species differentially expressed in DF granulosa cells, but only 8 of these encode proteins known to be involved in DF development. Most characterised transcripts upregulated in the DF are from tissue development genes that regulate cell differentiation, proliferation, apoptosis, signalling and tissue remodelling. Semiquantitative real-time PCR analysis confirmed seven genes with upregulated (P%0.05) mRNA expression in DF compared with both COH and SF granulosa cells. Thus, the new genes identified by SAGE and real-time PCR, which show enhanced mRNA expression in the DF, may regulate proliferation (cyclin D2; CCND2), prevention of apoptosis or DNA damage (growth arrest and DNA damage-inducible, b; GADD45B), RNA synthesis (splicing factor, arginine/serine rich 9; SFRS9) and unknown processes associated with enhanced steroidogenesis (ovary-specific acidic protein; DQ004742) in granulosa cells of DF at the onset of LH-dependent development. Further studies are required to show whether the expression of identified genes is dysregulated when abnormalities occur during DF selection or subsequent development.

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Identification of Developmentally Regulated Genes in the Somatic Cells of the Mouse Testis Using Serial Analysis of Gene Expression1

Cited by 44 publications

References 68 publications

Bestrophin 1 is indispensable for volume regulation in human retinal pigment epithelium cells

Bestrophin 1 is indispensable for volume regulation in human retinal pigment epithelium cells

Transcriptome profiling of the developing postnatal mouse testis using next-generation sequencing

Differentiation of the bovine dominant follicle from the cohort upregulates mRNA expression for new tissue development genes

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