216. c-kit Expression study: timing of onset in rodent testis and irradiated rat testis model

2005 ◽  
Vol 17 (9) ◽  
pp. 83
Author(s):  
S. Mithra Prabhu ◽  
M. L. Meistrich ◽  
S. Mendis ◽  
E. A. McLaughlin ◽  
K. L. Loveland
Keyword(s):  
Protein Expression ◽  
Germ Cell ◽  
Somatic Cell ◽  
Messenger Rna ◽  
Cell Function ◽  
Primordial Germ Cell ◽  
In Situ Hybridisation ◽  
Hormone Treatment ◽  
Normal Male ◽  
Rat Testis

Primordial germ cell and spermatogonial cell function is essential for normal male fertility. These cells require Sertoli–germ cell interactions, specifically somatic cell-derived stem cell factor (SCF) that acts through the c-kit receptor to govern primordial germ cell migration in the foetus, spermatogonial differentiation during puberty and adulthood, and Leydig cell steroidogenesis. We performed a comprehensive study of the c-kit mRNA expression profile in the pre- and post-pubertal mouse and rat testes by in situ hybridisation. Expression of c-kit mRNA was first visualised in germ cells after birth, with the levels concordant with the number and appearance of the differentiated spermatogonial subtypes in both the rat and the mouse. We also studied c-kit expression in the irradiated adult rat testis, in which only undifferentiated spermatogonia are present. After treatment with Cetrorelix, GnRH antagonist (3 days, 1, 2 and 4 weeks) germ cell maturation is re-initiated. Expression of c-kit messenger RNA was observed in the undifferentiated spermatogonia in both untreated and treated testes sections. In contrast, c-kit protein expression was undetectable until 4 weeks of hormone treatment. This suggests that c-kit mRNA and protein expression are differentially regulated and that protein expression relates to somatic cell function.

Sall4 Is Essential for Mouse Primordial Germ Cell Specification by Suppressing Somatic Cell Program Genes

Stem Cells ◽  
2014 ◽  
Vol 33 (1) ◽  
pp. 289-300 ◽  
Author(s):  
Yasuka L. Yamaguchi ◽  
Satomi S. Tanaka ◽  
Maho Kumagai ◽  
Yuka Fujimoto ◽  
Takeshi Terabayashi ◽  
...  
Keyword(s):  
Germ Cell ◽  
Somatic Cell ◽  
Primordial Germ Cell ◽  
Cell Specification ◽  
Germ Cell Specification ◽  
Mouse Primordial Germ Cell

234. Hedgehog signalling components in adult rat testis spermatogonial cells

2008 ◽  
Vol 20 (9) ◽  
pp. 34
Author(s):  
Z. Sahin ◽  
M. Meistrich ◽  
A. Szczepny ◽  
K. Loveland
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
In Situ Hybridisation ◽  
Cell Types ◽  
Spermatogonial Stem Cells ◽  
Male Rats ◽  
Brown Norway ◽  
Hedgehog Signalling ◽  
Rat Testis ◽  
Adult Rat

In normal tissues, Hedgehog-induced progenitor cell proliferation is transient and tightly regulated, preventing continuous regeneration. However, activation of constitutive Hedgehog signalling results in unregulated self-renewal of progenitor cells in association with several human cancers. Although the contribution of Hedgehog signalling to cancers is widely accepted, its impact on spermatogonial stem cells and impact on male fertility are unknown. In this study, we aimed to clarify the possible role of Hh signalling on normal spermatogenesis in the adult rat and in adult testicular stem cells in the irradiated model {1}. Adult male rats were obtained from Monash University Central Animal Service and killed by cervical dislocation before tissue removal and fixation in Bouins for routine histochemical procedures. For studies on irradiated testes, adult LBNF1 male rats (hybrids between Lewis and Brown–Norway) were purchased from Harlan Sprague–Dawley, Inc. (Indianapolis, IN, USA). Testes were irradiated with 6 Gy to deplete all maturing germ cell types. At 15 weeks after irradiation the animals were injected simultaneously with 1.5 mg each of Cetrorelix pamoate and Cetrorelix acetate. Testes were collected 1, 2 or 4 weeks after injection. In situ hybridisation combined with immunohistochemistry was performed using DIG-labelled cRNA probes to identify the cells in which Hedgehog signalling components are made {2}. Signals for mRNAs encoding t he transmembrane receptors Ptc2 and Smo are most intensely detected in spermatogonia and spermatocytes and are much less intense in the round spermatids. The mRNA for the cytoplamic regulator, Fused, is restricted to the earliest germ cell types, whereas expression of the negative cytoplasmic regulator, SuFu, only begins in the round spermatids and persists in elongating spermatids. Gli1 and Gli3 are expressed from spermatogonia through to round spermatids, whereas Gli2 is restricted to spermatogonia and spermatocytes. This pattern mimics what was reported for mouse {2}. Examination of the irradiated rat testis model revealed that Hedgehog signalling machinery is produced by resting spermatogonial stem cells but is turned off when they differentiate in response to hormones. This matches the emerging understanding of Hedgehog signals in cancer stem cells and provides the first demonstration that Hedgehog signalling may influence stem cells in the adult testis. (1) Shuttlesworth G.A. et al. 2000. Endocrinology. 141: 37–49 (2) Szczepny A. et al. 2006. Dev Dyn. 235:3063–3070.

Solea senegalensis vasa transcripts: molecular characterisation, tissue distribution and developmental expression profiles

2013 ◽  
Vol 25 (4) ◽  
pp. 646 ◽  
Author(s):  
Tiziana Pacchiarini ◽  
Ismael Cross ◽  
Ricardo B. Leite ◽  
Paulo Gavaia ◽  
Juan B. Ortiz-Delgado ◽  
...  
Keyword(s):  
Germ Cell ◽  
Expression Profiles ◽  
Primordial Germ Cell ◽  
In Situ Hybridisation ◽  
Developmental Expression ◽  
Solea Senegalensis ◽  
Senegalese Sole ◽  
Gonad Differentiation ◽  
Vasa Gene ◽  
Germinal Cells

The Vasa protein is an RNA helicase belonging the DEAD (Asp-Glu-Ala-Asp)-box family. The crucial role played by the vasa gene in the germ-cell lineage of both vertebrates and invertebrates has made this gene a useful molecular marker for germinal cells and a useful tool in surrogate broodstock production using primordial germ cell transplantation. With the aim of establishing a novel approach to improving Solea senegalensis broodstock management, the vasa gene in this species was characterised. Four S. senegalensis vasa transcripts were isolated: Ssvasa1, Ssvasa2, Ssvasa3 and Ssvasa4. Their phylogenetic relationship with other vasa homologues was determined confirming the high degree of conservation of this helicase throughout evolution. Our qPCR results showed that S. senegalensis vasa transcripts are prevalently expressed in gonads, with ovary-specific expression for Ssvasa3 and Ssvasa4. During embryonic and larval development, a switch between the longest and the shortest transcripts was observed. While Ssvasa1 and Ssvasa2 were maternally supplied, Ssvasa3 and Ssvasa4 depended on the de novo expression program of the growing juveniles, suggesting that vasa mRNA could be involved in Senegalese sole gonad differentiation. In situ hybridisation and immunohistochemical analysis performed in 150-days after hatching (DAH) larvae showed vasa product expression in the germinal region of early gonads. In our work we demonstrated the usefulness of Ssvasa mRNAs as molecular markers for primordial germ cells and germinal cells during embryonic development, larval ontogenesis and gonad differentiation. Furthermore, our results confirmed the potential of vasa to help investigate germinal cell biotechnology for Senegalese sole reproduction.

scholarly journals Induction of meiosis by embryonic gonadal somatic cells differentiated from pluripotent stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Haiying Wang ◽  
Linlin Liu ◽  
Chang Liu ◽  
Lingling Wang ◽  
Jiyu Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Germ Cell ◽  
Small Molecules ◽  
Somatic Cell ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cell ◽  
Somatic Cells ◽  
Embryonic Stem

Abstract Background Depletion of oocytes leads to ovarian aging-associated infertility, endocrine disruption and related diseases. Excitingly, unlimited oocytes can be generated by differentiation of primordial germ cell like cells (PGCLCs) from pluripotent stem cells. Nevertheless, development of oocytes and follicles from PGCLCs relies on developmentally matched gonadal somatic cells, only available from E12.5 embryos in mice. It is therefore imperative to achieve an in vitro source of E12.5 gonadal somatic cells. Methods We explored to identify small molecules, which can induce female embryonic stem cells (ESCs) into gonadal somatic cell like cells. Results Using RNA-sequencing, we identified signaling pathways highly upregulated in E12.5_gonadal somatic cells (E12.5_GSCs). Through searching for the activators of these pathways, we identified small-molecule compounds Vitamin C (Vc) and AM580 in combination (V580) for inducing differentiation of female embryonic stem cells (ESCs) into E12.5_GSC-like cells (E12.5_GSCLCs). After V580 treatment for 6 days and sorted by a surface marker CD63, the cell population yielded a transcriptome profile similar to that of E12.5_GSCs, which promoted meiosis progression and folliculogenesis of primordial germ cells. This approach will contribute to the study of germ cell and follicle development and oocyte production and have implications in potentially treating female infertility. Conclusion ESCs can be induced into embryonic gonadal somatic cell like cells by small molecules.

Germ-cell transfer and sex ratio in Xenopus laevis

Development ◽  
1965 ◽  
Vol 13 (1) ◽  
pp. 51-61
Author(s):  
A. W. Blackler
Keyword(s):  
Xenopus Laevis ◽  
Germ Cell ◽  
South African ◽  
Germ Cells ◽  
Sexual Differentiation ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Hormone Treatment ◽  
Sex Reversal ◽  
Cell Transfer

A Technique for the transfer of primordial germ cells between neurulae of the South African Clawed Toad Xenopus laevis has been described by Blackler & Fischberg (1961). This method was originally developed with the object in mind of eventually making a genetic analysis of abnormal embryos resulting from the transplantation of somatic nuclei. Such analysis involves two schemes which require the transfer of embryonic gonocytes from the defective transplant embryo to a normal recipient. Moreover, one of these two schemes requires that transferred germ cells be reversed in their sexual differentiation in the developing gonad of the host (see Fischberg, 1961; Fischberg & Blackler, 1963a, b). Since it has been known for some time, from experiments involving parabiosis, transplantation of the gonadal rudiment and hormone treatment (e.g. Burns, 1925, 1930; Witschi, 1937; Humphrey, 1929, 1933, 1948, 1957; Gallien, 1953, 1956), that the manifestation of the sex genotype of a primordial germ cell can be physiologically reversed by the hormonal characteristics of the gonad, there seemed no obstacle to obtaining sex-reversal of the transferred gonocytes in Xenopus.

Primordial germ cell-somatic cell partnership: A balancing cell signaling act

2001 ◽  
Vol 60 (3) ◽  
pp. 277-280 ◽  
Author(s):  
Abraham L. Kierszenbaum ◽  
Laura L. Tres
Keyword(s):  
Cell Signaling ◽  
Germ Cell ◽  
Somatic Cell ◽  
Primordial Germ Cell

scholarly journals Effect of germ cell depletion on levels of specific mRNA transcripts in mouse Sertoli cells and Leydig cells

Reproduction ◽  
2008 ◽  
Vol 135 (6) ◽  
pp. 839-850 ◽  
Author(s):  
P J O'Shaughnessy ◽  
L Hu ◽  
P J Baker
Keyword(s):  
Germ Cell ◽  
Somatic Cell ◽  
Sertoli Cell ◽  
Germ Cells ◽  
Leydig Cell ◽  
Cell Function ◽  
Cell Activity ◽  
Cell Depletion ◽  
Transcript Levels ◽  
Mrna Transcripts

It has been shown that testicular germ cell development is critically dependent upon somatic cell activity but, conversely, the extent to which germ cells normally regulate somatic cell function is less clear. This study was designed, therefore, to examine the effect of germ cell depletion on Sertoli cell and Leydig cell transcript levels. Mice were treated with busulphan to deplete the germ cell population and levels of mRNA transcripts encoding 26 Sertoli cell-specific proteins and 6 Leydig cell proteins were measured by real-time PCR up to 50 days after treatment. Spermatogonia were lost from the testis between 5 and 10 days after treatment, while spermatocytes were depleted after 10 days and spermatids after 20 days. By 30 days after treatment, most tubules were devoid of germ cells. Circulating FSH and intratesticular testosterone were not significantly affected by treatment. Of the 26 Sertoli cell markers tested, 13 showed no change in transcript levels after busulphan treatment, 2 showed decreased levels, 9 showed increased levels and 2 showed a biphasic response. In 60% of cases, changes in transcript levels occurred after the loss of the spermatids. Levels of mRNA transcripts encoding Leydig cell-specific products related to steroidogenesis were unaffected by treatment. Results indicate (1) that germ cells play a major and widespread role in the regulation of Sertoli cell activity, (2) most changes in transcript levels are associated with the loss of spermatids and (3) Leydig cell steroidogenesis is largely unaffected by germ cell ablation.

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