scholarly journals In vitro Spermatogenesis – Optimal Culture Conditions for Testicular Cell Survival, Germ Cell Differentiation, and Steroidogenesis in Rats

2014 ◽  
Vol 5 ◽  
Author(s):  
Ahmed Reda ◽  
Mi Hou ◽  
Luise Landreh ◽  
Kristín Rós Kjartansdóttir ◽  
Konstantin Svechnikov ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Germ Cell ◽  
Cell Survival ◽  
Culture Conditions ◽  
Testicular Cell ◽  
In Vitro Spermatogenesis ◽  
Germ Cell Differentiation

scholarly journals In Vitro Germ Cell Differentiation from Cynomolgus Monkey Embryonic Stem Cells

PLoS ONE ◽  
2009 ◽  
Vol 4 (4) ◽  
pp. e5338 ◽  
Author(s):  
Kaori Yamauchi ◽  
Kouichi Hasegawa ◽  
Shinichiro Chuma ◽  
Norio Nakatsuji ◽  
Hirofumi Suemori
Keyword(s):  
Stem Cells ◽  
Cell Differentiation ◽  
Embryonic Stem Cells ◽  
Germ Cell ◽  
Cynomolgus Monkey ◽  
Embryonic Stem ◽  
Germ Cell Differentiation

Role of Mael in early oogenesis and during germ-cell differentiation from embryonic stem cells in mice in vitro

Zygote ◽  
2013 ◽  
Vol 22 (4) ◽  
pp. 513-520 ◽  
Author(s):  
I. Bahena ◽  
E. Xu ◽  
M. Betancourt ◽  
E. Casas ◽  
Y. Ducolomb ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Differentiation ◽  
Embryonic Stem Cells ◽  
Germ Cell ◽  
Embryonic Stem ◽  
Oocyte Growth ◽  
Germ Cell Differentiation ◽  
Rnai Technology

SummaryIn a previous study, we have identified a set of conserved spermatogenic genes whose expression is restricted to testis and ovary and that are developmentally regulated. One of these genes, the transcription factor Mael, has been reported to play an essential role in mouse spermatogenesis. Nevertheless, the role of Mael in mouse oogenesis has not been defined. In order to analyse the role of Mael in mouse oogenesis, the expression of this gene was blocked during early oogenesis in mouse in vitro using RNAi technology. In addition, the role of Mael during differentiation of embryonic stem cells (ESC) into germ cells in vitro was analysed. Results show that downregulation of Mael by a specific short interfering RNA disrupted fetal oocyte growth and differentiation in fetal ovary explants in culture and the expression of several germ-cell markers in ESC during their differentiation. These results suggest that there is an important role for Mael in early oogenesis and during germ-cell differentiation from embryonic stem cells in mouse in vitro.

167. MUSASHI FAMILY OF RNA BINDING PROTEINS: CELL CYCLE REGULATORS IN SPERMATOGENESIS

2010 ◽  
Vol 22 (9) ◽  
pp. 85
Author(s):  
E. A. McLaughlin ◽  
B. A. Fraser ◽  
V. Pye ◽  
M. Bigland ◽  
N. A. Siddall ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Differentiation ◽  
Germ Cell ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Translational Repression ◽  
Germ Cell Differentiation ◽  
Pachytene Spermatocytes

Mammalian meiosis is a tightly regulated process involving specialized cell cycle progression and morphogenetic changes. We have demonstrated that the Musashi family of RNA binding proteins is implicated in the regulation of spermatogonial stem self renewal and germ cell differentiation. Here we describe the novel mechanism by which the Musashi family proteins, Msi1 and Msi2, act to control exit from spermatogonial mitotic amplification and normal entry into meiosis. Gene and protein analysis indicated overlapping Msi1 and Msi2 profiles in enriched populations of isolated germ cells and reciprocal subcellular expression patterns in spermatogonia and pachytene spermatocytes/ round spermatids in testes sections. Recombinant Msi1 protein-RNA pulldown and microarray analysis coupled with in vitro shRNA knockdown studies in spermatogonial culture and subsequent immunoprecipitation and qPCR established that Msi1 targeted Msi2 mRNA for post transcriptional translational repression. Immunoprecipitation of Msi2 target mRNA and subsequent qPCR together with in vitro shRNA knockdown studies inround spermatidculture identified a cell cycle inhibitor protein CDKN1C (p57kip2) as the principal target of Msi2 translational inhibition. Immunolocalisation of CDKN1C protein indicated that expression of this cell cycle regulator coincided with the nuclear import of Msi1 and the appearance of cytoplasmic Msi2 expression in early pachytene spermatocytes. Using a transgenic Msi1 overexpression mouse model in conjunction with quantitative gene and protein expression, we confirmed Msi1 targeting of Msi2 and subsequent Msi2 targeting of CDKN1C for translational repression in vivo. Ectopic overexpression of Msi1 in germ cellsinduces substantial Msi2 downregulation and aberrant CDKN1C expression, resulting in abnormal spermatogenic differentiation, germ cell apoptosis/arrest and sterility. In conclusion, our results indicate a sophisticated molecular switch encompassing cell cycle protein regulation by Musashi family proteins, is required for normal exit from mitotic division, entry into meiosis and post meiotic germ cell differentiation.

Hormones and the differentiation of type A spermatogonia in mouse cryptorchid testes incubated in vitro

1982 ◽  
Vol 94 (1) ◽  
pp. 43-NP ◽  
Author(s):  
Tatsuji Haneji ◽  
Yoshitake Nishimune
Keyword(s):  
Cell Differentiation ◽  
Germ Cell ◽  
Cyclic Adenosine Monophosphate ◽  
Type A ◽  
Adenosine Monophosphate ◽  
Testicular Germ Cell ◽  
Adult Mice ◽  
Germ Cell Differentiation ◽  
Type A Spermatogonia

In order to study hormonal effects on testicular germ cell differentiation, especially on type A spermatogonia, artificially induced cryptorchid testes of adult mice were cultured in a medium containing testosterone, dihydrotestosterone, tri-iodothyronine, dibutyryl 3′: 5′ cyclic adenosine monophosphate, human chorionic gonadotrophin, LH, FSH, insulin and transferrin. These substances, with the exception of FSH, showed no stimulatory effect on the differentiation of type A spermatogonia. However, FSH activated cell division in type A spermatogonia and stimulated them to differentiate, while LH showed neither the promotion of differentiation nor a synergistic effect on FSH-mediated germ cell differentiation.

scholarly journals Critical role of Emx2 in the pluripotency – differentiation transition in male gonocytes via regulation of FGF9/NODAL pathway

Reproduction ◽  
2016 ◽  
Vol 151 (6) ◽  
pp. 673-681 ◽  
Author(s):  
Ma Tian-Zhong ◽  
Chen Bi ◽  
Zhang Ying ◽  
Jing Xia ◽  
Peng Cai-Ling ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Mouse Model ◽  
Germ Cell ◽  
Germ Cells ◽  
Critical Role ◽  
Genital Ridge ◽  
Germ Cell Differentiation ◽  
Pluripotency Markers

Abstract Emx2 deletion impairs the growth and maintenance of the genital ridge. However, its role in subsequent germ cell differentiation during embryonic stages is unknown. Using a tamoxifen-inducible Cre-loxP mouse model (Emx2flox/flox, Cre-ERTM, hereafter called as Emx2 knockdown), we showed that germ cell differentiation was impaired in Emx2-knockdown testes. Representative characteristics of male germ cell differentiation, including a reduced ability to form embryonic germ (EG) cell colonies in vitro, down-regulation of pluripotency markers and G1/G0 arrest, did not occur in Emx2-knockdown testes. Furthermore, FGF9 and NODAL signalling occurred at abnormally high levels in Emx2-knockdown testes. Both blocking FGF9 signalling with SU5402 and inhibiting NODAL signalling with SB431542 allowed germ cells from Emx2-knockdown testes to differentiate in vitro. Therefore, EMX2 in somatic cells is required to trigger germ cell differentiation in XY foetuses, posterior to its previously reported role in the growth and maintenance of the genital ridge.

scholarly journals Hedgehog signalling promotes germ cell survival in the rat testis

Reproduction ◽  
2011 ◽  
Vol 142 (5) ◽  
pp. 711-721 ◽  
Author(s):  
Juho-Antti Mäkelä ◽  
Vuokko Saario ◽  
Sonia Bourguiba-Hachemi ◽  
Mirja Nurmio ◽  
Kirsi Jahnukainen ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Germ Cell ◽  
Germ Cells ◽  
Tyrosine Kinases ◽  
Negative Regulator ◽  
Mrna Levels ◽  
Paracrine Signalling ◽  
Rat Testis ◽  
Germ Cell Differentiation

Hedgehog (Hh) signalling has a crucial role in testis development. Sertoli cell-derived desert hedgehog (DHH) guides the formation of testis cords and differentiation of foetal-type Leydig cells. Dhh mutant mice are infertile due to a block in germ cell differentiation, hypogonadism and hypoandrogenism. Hh signalling pathway components are also expressed in postnatal testis. In the rat testis the transcription factor of the Hh pathway, glioma-associated oncogene homologue (GLI1), is expressed by a wide variety of germ cells. This suggests that Hh signalling is involved in spermatogenesis at many different levels. Our data show that canonical Hh signalling is turned off in early condensing spermatids that strongly express the negative regulator of the pathway, suppressor of fused (SUFU). Most of the Hh pathway specific mRNAs display the highest values in stages II–VI of the rat seminiferous epithelial cycle. The key endocrine regulator of germ cell differentiation, FSH, down-regulates Dhh mRNA levels in vitro. Hh signalling inhibition in vitro leads to massive apoptosis of germ cells. In prepubertal rat testis imatinib mesylate-induced inhibition of tyrosine kinases impinges on Dhh transcript levels and Hh signalling. Our data indicate that Hh signalling is part of the paracrine signalling network in the rat testis. It promotes the survival of germ cells and is suppressed by FSH.

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