Analysis of mouse germ-cell transcriptome at different stages of spermatogenesis by SAGE: Biological significance

Wu, Shao-Ming; Baxendale, Vanessa; Chen, Yali; Pang, Alan Lap‐Yin; Stitely, Timothy; Munson, Peter J.; Leung, Mason C.P.; Ravindranath, Neelakanta; Dym, Martin; Rennert, Owen M.; Chan, Wai‐Yee

doi:10.1016/j.ygeno.2004.08.018

Cited by 53 publications

(46 citation statements)

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“…In addition, the testis is a very complicated structure with duct cells, germ cells, juvenile sperm, and support cells for the spermatogonia and endocrine cells, and the high percentage of genes expressed in testis may partially reflect the complexity of the organ. Our finding of a high percentage of genes expressed in the testis is consistent with the report by Wu et al (2004). The LacZ screen may contribute to identifying those genes involved in stem-cell maintenance and unique to the development of germ plasm since ∼8% of all genes surveyed were exclusively expressed in the testis.…”

Section: Discussionsupporting

confidence: 90%

A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

et al. 2015

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Expression of the bacterial beta-galactosidase reporter gene (lacZ) in the vector used for the Knockout Mouse Project (KOMP) is driven by the endogenous promoter of the target gene. In tissues from KOMP mice, histochemical staining for LacZ enzyme activity can be used to determine gene expression patterns. With this technique, we have produced a comprehensive resource of gene expression using both whole mount (WM) and frozen section (FS) LacZ staining in 313 unique KOMP mutant mouse lines. Of these, ∼80% of mutants showed specific staining in one or more tissues, while ∼20% showed no specific staining, ∼13% had staining in only one tissue, and ∼25% had staining in >6 tissues. The highest frequency of specific staining occurred in the brain (∼50%), male gonads (42%), and kidney (39%). The WM method was useful for rapidly identifying whole organ and some substructure staining, while the FS method often revealed substructure and cellular staining specificity. Both staining methods had >90% repeatability in biological replicates. Nonspecific LacZ staining occurs in some tissues due to the presence of bacteria or endogenous enzyme activity. However, this can be effectively distinguished from reporter gene activity by the combination of the WM and FS methods. After careful annotation, LacZ staining patterns in a high percentage of mutants revealed a unique structure-function not previously reported for many of these genes. The validation of methods for LacZ staining, annotation, and expression analysis reported here provides unique insights into the function of genes for which little is currently known.[Supplemental material is available for this article.]

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Section: Discussionsupporting

confidence: 90%

A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

et al. 2015

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“…We demonstrated, using microarray analysis, that Col1a1 and Col1a2 were abundantly expressed in the seminiferous tubules of immature mice; however, their mRNA expression was apparently at lower levels in the seminiferous tubules of adult mice. The microarray data were corroborated by a semi-quantitative RT-PCR assay and are also in agreement with our group's recent findings that Col1a2 was among the most abundant type A spermatogonial-specific transcripts as seen by the use of serial analysis of gene expression (SAGE) (Wu et al 2004). In the testis of 6-day-old mice, seminiferous tubules consist of only one type of germ cell, namely primitive type A spermatogonia, and the differentiation of spermatogonia is barely initiated (Dettin et al 2003).…”

Section: Discussionsupporting

confidence: 90%

Expression of Col1a1, Col1a2 and procollagen I in germ cells of immature and adult mouse testis

He¹,

Feng²,

Xiao-dong³

et al. 2005

Reproduction

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The objective of this study was to compare the expression of Col1a1, Col1a2, and procollagen I in the seminiferous tubules of immature and adult mice and to characterize the cellular expression pattern of procollagen I in germ cells during spermatogenesis in order to provide necessary groundwork for further functional studies in the process of spermatogenesis. Microarray analysis demonstrated that Col1a1 and Col1a2 were abundantly expressed in the seminiferous tubules of 6-day-old mice compared with 60-day-old mice, and the expression levels of Col1a1 and Col1a2 mRNA were validated using a semi-quantitative RT-PCR assay. Western blot analysis further confirmed that procollagen I was expressed at a higher level in the seminiferous tubules of 6-day-old mice compared with 60-day-old mice. Immunohistochemical analysis revealed that type A spermatogonia were positive for procollagen I in the testis of 6-day-old mice, whereas Sertoli cells were negative for this protein. The in vivo procollagen I staining in type A spermatogonia was corroborated in spermatogonia exhibiting a high potential for proliferation and the ability to form germ cell colonies in in vitro culture. Moreover, procollagen I was also detected in type A spermatogonia, intermediate spermatogonia, type B spermatogonia, and preleptotene spermatocytes in the adult mouse testes, but positive staining disappeared in more differentiated germ cell lineages detaching from the basement membrane, including leptotene spermatocytes, pachytene spermatocytes, round spermatids and elongated spermatids. These data suggest that Col1a1, Col1a2 and procollagen I are associated with type A spermatogonia and play a potential role in mediating the detachment and migration of germ cells during spermatogenesis.

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“…They identified 12976 unique transcripts and found that most of the Sertoli or Leydig cell-specific genes are up-regulated in adult mice. Wu et al [14] sequenced the transcriptomes of mouse type A spermatogonia, pachytene spermatocytes, and round spermatids and, again using SAGE, estimated that the germ-cell transcriptome was comprised of more than 30000 transcripts. Yao et al [15] identified 19323 unique tags from both young adult and aged mouse testes.…”

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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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Analysis of mouse germ-cell transcriptome at different stages of spermatogenesis by SAGE: Biological significance

Cited by 53 publications

References 50 publications

A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

A lacZ reporter gene expression atlas for 313 adult KOMP mutant mouse lines

Expression of Col1a1, Col1a2 and procollagen I in germ cells of immature and adult mouse testis

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

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