Proteomic analysis of the spermatogonial stem cell compartment in dogfish Scyliorhinus canicula L

Loppion, Géraldine; Lavigne, Régis; Pineau, Charles; Auvray, Pierrı̈ck; Sourdaine, Pascal

doi:10.1016/j.cbd.2010.03.004

Cited by 7 publications

(6 citation statements)

References 29 publications

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“…Therefore, undifferentiated spermatogonia were found to be distributed from stage I (germinative zone) to stage II, which could correspond to the successive types of A s , A p , and A al described for rodents. Moreover, the phylogenetic position of Chondrichthyans, the progress in sequence database production for several elasmobranchs (Coolen et al 2007, Venkatesh et al 2007, King et al 2011, Tan et al 2012, Quan et al 2013, and the possibility to perform transcriptome (Redon et al 2010) and proteome (Loppion et al 2010) analyses makes the dogfish an excellent reference species to study and understand the evolution of the SSC niche.…”

Section: Introductionmentioning

confidence: 99%

Characterization of spermatogonial markers in the mature testis of the dogfish (Scyliorhinus canicula L.)

Bosseboeuf¹,

Gautier²,

Auvray³

et al. 2014

Reproduction

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In dogfish, spermatogenesis progresses from a restricted germinative zone, which lines the dorsal testicular vessel. Single spermatogonia (A s ), including the spermatogonial stem cells (SSCs), produce successively paired (A p ), undifferentiated (A u4 to A u512 ), and differentiated (A d1 to A d8 ) spermatogonia and preleptotene (PL) spermatocytes through 13 mitoses. Dogfish spermatogonial subpopulations present classical morphological characteristics but cannot be distinguished on the basis of molecular markers. This characterization has been initiated in mammals despite the difficulty to separate each spermatogonial subpopulation. For instance, both glial cell-derived neurotrophic factor family receptor alpha 1 (GFRa1) and promyelocytic leukemia zinc finger protein (PLZF) are markers of undifferentiated spermatogonia, whereas receptor tyrosine kinase C-kit is a marker of differentiated spermatogonia. The aim of this study is to characterize spermatogonial markers and to differentiate several spermatogonial subpopulations. Dogfish cDNA sequences have been identified and validated by phylogenetic analyses for gfra1, plzf, pou2, as well as for high-mobility group box proteins 2 and 3 (hmgb2 and 3) and for mini-chromosome maintenance protein 6 (mcm6). We have used the anatomical advantage of the polarized dogfish testis to analyze the expression of those markers by RT-PCR and in situ hybridization. gfra1, pou2, and plzf have been detected in the testicular germinative zone, suggesting that spermatogonial markers are relatively well conserved among vertebrates but with a less restricted expression for plzf. Moreover, hmgb3 and mcm6 have been identified as new markers of differentiated spermatogonia. Finally, this first molecular characterization of spermatogonial subpopulations in a chondrichthyan model will be useful for further studies on the SSC niche evolution.

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Section: Introductionmentioning

confidence: 99%

Characterization of spermatogonial markers in the mature testis of the dogfish (Scyliorhinus canicula L.)

Bosseboeuf¹,

Gautier²,

Auvray³

et al. 2014

Reproduction

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“…2D-gels have been the proteomic tool of choice for studies of germline stem cells in chickens [21], in germline and embryonic SCs in mice [18] and of rat spermatogonia [41] revealing 56, 166 and 102 proteins respectively. More recently, the spermatogonial stem cell niche of dogfish has been examined yielding 16 protein identifications [22]. In this study, we report the comprehensive proteomic profiling of bovine testis cell populations which enabled the identification of 1,387 proteins, of which 1,150 were identified in the population enriched for undifferentiated spermatogonia.…”

Section: Discussionmentioning

confidence: 99%

“…Improvements in sensitivity, accuracy and speed of mass spectrometric analysis when combined with automated methods for data acquisition and processing have enabled protein profiling experiments in a high throughput manner. Over recent years, proteomic studies have been applied to the study of SSC from rats [16], mice [17][18][19], humans [20], chickens [21] and dogfish [22]. Proteomics can reveal information about protein expression (presence and quantity) and post-translational modification which facilitates our understanding of protein interactions and function in complex systems.…”

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confidence: 99%

Global proteomic profiling of the membrane compartment of bovine testis cell populations

Colgrave

Stockwell²,

Grace³

et al. 2013

JIOMICS

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1. Introduction Spermatogonial stem cells hold enormous potential in mammalian reproductive medicine through the preservation of gametes, the restoration of fertility, enhancement of germ-lineage genetic manipulation and the improvement in our understanding of stem cell biology. Here we describe the protein profiles of the membrane compartment of bovine testicular cell isolates which were enriched for germ cells using differential plating. The isolated cells were characterised with antibodies to UCHL1 (previously known as PGP9.5) for type A spermatogonia; DDX4 (previously known as VASA) for germ cells and vimentin for Sertoli cells. Ultracentrifugation techniques were used to specifically isolate cell membranes, with membrane protein identifications significantly increased when compared to whole cell lysates. We utilised the filter-aided sample preparation protocol for improved digestion efficiency of membrane proteins. Using ESI-LC-MS/MS, we compared the proteins present in two cell populations. A total of 1,387 proteins were identified in bovine testis cell isolates, of which 39% were membraneassociated. A total of 64 proteins were differentially expressed in the non-adhered fraction (enriched for undifferentiated germ cells) compared to the adhered fraction, of which 16 were unique to this cell population and the remaining 48 showed a twofold change (increase when compared to the adhered cell population) as judged by spectral counting. This analysis revealed a number of candidate germ cell markers including the known markers, DDX4 and UCHL1. The proteomic profiles generated in this study support and complement transcription data on gene expression and histological levels, and reinforce the potential of proteomics in identifying and characterising the protein effectors of self-renewal and/or differentiation in stem cells.

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“…22 A different approach to characterize the proteome of the spermatogonial stem cell exploited the peculiarities of the testis developmental biology in the dogfish Scyliorhinus canicula. 54 These authors isolated, under stereomicroscope and dissection, the testicular germinative zone, highly enriched in spermatogonial stem cells, and used 2D and MALDI-TOF/TOF to identify 16 proteins and to also demonstrate the feasibility of this model to study the stem cell niche. 54 Still, a different set of studies has used testicular cell sorting to obtain enriched cellular fractions with which proteomic analysis is performed.…”

Section: Testicular Proteomics: Spermatogonial Stem Cells Spermatocymentioning

confidence: 99%

“…54 These authors isolated, under stereomicroscope and dissection, the testicular germinative zone, highly enriched in spermatogonial stem cells, and used 2D and MALDI-TOF/TOF to identify 16 proteins and to also demonstrate the feasibility of this model to study the stem cell niche. 54 Still, a different set of studies has used testicular cell sorting to obtain enriched cellular fractions with which proteomic analysis is performed. This approach was applied to separate spermatogonia from 9-day-old rats followed by protein 2D analysis and identification of the proteins using MALDI-TOF.…”

Section: Testicular Proteomics: Spermatogonial Stem Cells Spermatocymentioning

confidence: 99%

Proteomics and the genetics of sperm chromatin condensation

Oliva

Castillo²

2010

Asian J Androl

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Spermatogenesis involves extremely marked cellular, genetic and chromatin changes resulting in the generation of the highly specialized sperm cell. Proteomics allows the identification of the proteins that compose the spermatogenic cells and the study of their function. The recent developments in mass spectrometry (MS) have markedly increased the throughput to identify and to study the sperm proteins. Catalogs of thousands of testis and spermatozoan proteins in human and different model species are becoming available, setting up the basis for subsequent research, diagnostic applications and possibly the future development of specific treatments. The present review intends to summarize the key genetic and chromatin changes at the different stages of spermatogenesis and in the mature sperm cell and to comment on the presently available proteomic studies.

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Proteomic analysis of the spermatogonial stem cell compartment in dogfish Scyliorhinus canicula L

Cited by 7 publications

References 29 publications

Characterization of spermatogonial markers in the mature testis of the dogfish (Scyliorhinus canicula L.)

Characterization of spermatogonial markers in the mature testis of the dogfish (Scyliorhinus canicula L.)

Global proteomic profiling of the membrane compartment of bovine testis cell populations

Proteomics and the genetics of sperm chromatin condensation

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