The In Vivo Response of Stem and Other Undifferentiated Spermatogonia to the Reversible Inhibition of Glial Cell Line-Derived Neurotrophic Factor Signaling in the Adult

Savitt, Joseph M.; Singh, Dolly; Zhang, Chao; Chen, Liang Chin; Folmer, Janet; Shokat, Kevan M.; Wright, William W.

doi:10.1002/stem.1028

Cited by 36 publications

(53 citation statements)

References 34 publications

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“…Additionally, 1NA-PP1 and 3MB-PP1 have been used to selectively target AS -alleles of Ret, EphB2, EphB3 and EphB4 in cells and mice despite weak inhibition of their WT -alleles in vitro 25, 26 . Thus, many of the WT -kinases identified as targets of 1NA-PP1, 1NM-PP1 or 3MB-PP1 in our screen and in the work of others are not likely to be bona fide targets in a cellular context.…”

Section: Resultsmentioning

confidence: 99%

Structure-Guided Inhibitor Design Expands the Scope of Analog-Sensitive Kinase Technology

et al. 2013

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Engineered analog-sensitive (AS) protein kinases have emerged as powerful tools for dissecting phospho-signaling pathways, for elucidating the cellular function of individual kinases, and for deciphering unanticipated effects of clinical therapeutics. A crucial and necessary feature of this technology is a bioorthogonal small molecule that is innocuous towards native cellular systems but can potently inhibit the engineered kinase. In order to generalize this method we sought a molecule capable of targeting divergent AS-kinases. Here we employ X-ray crystallography and medicinal chemistry to unravel the mechanism of current inhibitors and use these insights to design the most potent, selective and general AS-kinase inhibitors reported to date. We use large-scale kinase inhibitor profiling to characterize the selectivity of these molecules as well as examine the consequences of potential off-target effects in chemical genetic experiments. The molecules reported here will serve as powerful tools in efforts to extend AS-kinase technology to the entire kinome and the principles discovered may help in the design of other engineered enzyme/ligand pairs.

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Section: Resultsmentioning

confidence: 99%

Structure-Guided Inhibitor Design Expands the Scope of Analog-Sensitive Kinase Technology

et al. 2013

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“…3(D), EDN1 [cardiovascular and craniofacial development; (Chan et al, 2008)], GDNF (control of spermatogonial stem cell number; (Savitt et al, 2012); see Fig. 3(B), and RBP4 [maintenance of testis function during vitamin A starvation; (Ghyselinck et al, 2006)], as potentially relevant for the phenotype observed in the Sox8/Sox9 mutant.…”

Section: Identification Of Potential Direct Targets For Sox8 and Sox9mentioning

confidence: 99%

Genome‐wide identification of Sox8‐, and Sox9‐dependent genes during early post‐natal testis development in the mouse

et al. 2013

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Genome-wide identification of Sox8-, and Sox9-dependent genes during early post-natal testis development in the mouse F. Chalmel,*,a A. Lardenois,*,a I. Georg, †, ‡,k F. Barrionuevo, †,** P. Demougin, § B. J egou,*, ¶ G. Scherer † and M. Primig* *Inserm, U1085-Irset, University of Rennes 1, Rennes, France, †Institute of Human Genetics, University of Freiburg, Freiburg, Germany, ‡Faculty for Biology, University of Freiburg, Freiburg, Germany, §Biozentrum, University of Basel, Basel, Switzerland, ¶EHESP School of Public Health (Rennes, Paris), Rennes Cedex, France, k Present address: Area of Human DNA Variability, Centro de Genomica e Investigacio Oncologica (GENYO), Pfizer-Universidad de Granada-Junta de Andalucia, Granada, Spain, and **Departamento de Genetica e Instituto de Biotecnologia, Universidad de Granada, Armilla, Spain SUMMARYThe SOX8 and SOX9 transcription factors are involved in, among others, sex differentiation, male gonad development and adult maintenance of spermatogenesis. Sox8 À/À mice lacking Sox9 in Sertoli cells fail to form testis cords and cannot establish spermatogenesis. Although genetic and histological data show an important role for these transcription factors in regulating spermatogenesis, it is not clear which genes depend upon them at a genome-wide level. To identify transcripts that respond to the absence of Sox8 in all cells and Sox9 in Sertoli cells we measured mRNA concentrations in testicular samples from mice at 0, 6 and 18 days post-partum. In total, 621 and 629 transcripts were found at decreased or increased levels, respectively, at different time points in the mutant as compared to the control samples. These mRNAs were categorized as preferentially expressed in Sertoli cells or germ cells using data obtained with male and female gonad samples and enriched testicular cell populations. Five candidate genes were validated at the protein level. Furthermore, we identified putative direct SOX8 and SOX9 target genes by integrating predicted SOX-binding sites present in potential regulatory regions upstream of the transcription start site. Finally, we used protein network data to gain insight into the effects on regulatory interactions that occur when Sox8 and Sox9 are absent in developing Sertoli cells. The integration of testicular samples with enriched Sertoli cells, germ cells and female gonads enabled us to broadly distinguish transcripts directly affected in Sertoli cells from others that respond to secondary events in testicular cell types. Thus, combined RNA profiling signals, motif predictions and network data identified putative SOX8/SOX9 target genes in Sertoli cells and yielded insight into regulatory interactions that depend upon these transcription factors. In addition, our results will facilitate the interpretation of genome-wide in vivo SOX8 and SOX9 DNA binding data.

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“…line-derived neurotrophic factor, GDNF, (5)(6)(7); activin A and bone morphogenetic protein 4, BMP4, (8,9)], Leydig or peritubular myoid cells [colony-stimulating factor (CSF) 1, CSF1 (10 -11)], contribute to the niche homeostasis, and influence the balance between SSC self-renewal and differentiation (4). It is possible that yet unknown factors participate in regulating germ cell proliferation/ differentiation or that known factors exert yet unknown functions in the testis.…”

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confidence: 99%

Fsh Stimulates Spermatogonial Proliferation and Differentiation in Zebrafish via Igf3

et al. 2015

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Growth factors modulate germ line stem cell self-renewal and differentiation behavior. We investigate the effects of Igf3, a fish-specific member of the igf family. Fsh increased in a steroid-independent manner the number and mitotic index of single type A undifferentiated spermatogonia and of clones of type A differentiating spermatogonia in adult zebrafish testis. All 4 igf gene family members in zebrafish are expressed in the testis but in tissue culture only igf3 transcript levels increased in response to recombinant zebrafish Fsh. This occurred in a cAMP/protein kinase A-dependent manner, in line with the results of studies on the igf3 gene promoter. Igf3 protein was detected in Sertoli cells. Recombinant zebrafish Igf3 increased the mitotic index of type A undifferentiated and type A differentiating spermatogonia and up-regulated the expression of genes related to spermatogonial differentiation and entry into meiosis, but Igf3 did not modulate testicular androgen release. An Igf receptor inhibitor blocked these effects of Igf3. Importantly, the Igf receptor inhibitor also blocked Fsh-induced spermatogonial proliferation. We conclude that Fsh stimulated Sertoli cell production of Igf3, which promoted via Igf receptor signaling spermatogonial proliferation and differentiation and their entry into meiosis. Because previous work showed that Fsh also released spermatogonia from an inhibitory signal by down-regulating anti-Müllerian hormone and by stimulating androgen production, we can now present a model, in which Fsh orchestrates the activity of stimulatory (Igf3, androgens) and inhibitory (anti-Müllerian hormone) signals to promote spermatogenesis.

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The In Vivo Response of Stem and Other Undifferentiated Spermatogonia to the Reversible Inhibition of Glial Cell Line-Derived Neurotrophic Factor Signaling in the Adult

Cited by 36 publications

References 34 publications

Structure-Guided Inhibitor Design Expands the Scope of Analog-Sensitive Kinase Technology

Structure-Guided Inhibitor Design Expands the Scope of Analog-Sensitive Kinase Technology

Genome‐wide identification of Sox8‐, and Sox9‐dependent genes during early post‐natal testis development in the mouse

Fsh Stimulates Spermatogonial Proliferation and Differentiation in Zebrafish via Igf3

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