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Lili Ni, Hongchang Xie, Li Tan
The transcription factor FOXJ3 (Forkhead box J3) is highly expressed in spermatogonia and meiotic spermatocytes within mouse testes. Here, we addressed how FOXJ3 might participate in spermatogenesis using two conditional knockout mouse models in which the Foxj3 was deleted from either spermatogonia or meiotic spermatocytes. Both models exhibited complete male sterility, but distinct etiologies: Deleting FOXJ3 from spermatogonia using Foxj3(flox/flox) , Mvh-Cre mice caused Sertoli-cell-only syndrome in males...
October 14, 2016: Molecular Reproduction and Development
Neelam Pandey, Sarbani Giri, Samrat Das, Puja Upadhaya
Even though there are contradictory reports regarding the cellular and molecular changes induced by mobile phone emitted radiofrequency radiation (RFR), the possibility of any biological effect cannot be ruled out. In view of a widespread and extensive use of mobile phones, this study evaluates alterations in male germ cell transformation kinetics following RFR exposure and after recovery. Swiss albino mice were exposed to RFR (900 MHz) for 4 h and 8 h duration per day for 35 days. One group of animals was terminated after the exposure period, while others were kept for an additional 35 days post-exposure...
October 13, 2016: Toxicology and Industrial Health
F Kent Hamra
Knowledge gaps persist on signaling pathways and metabolic states in germ cells sufficient to support spermatogenesis independent of a somatic environment. Consequently, methods to culture mammalian stem cells through spermatogenesis in defined systems have not been established. Lack of success at culturing mammalian stem cells through spermatogenesis in defined systems reflects an inability to experimentally recapitulate biochemical events that develop in germ cells during a seminiferous epithelial cycle. Complex germ and somatic cell associations that develop each seminiferous epithelial cycle support such a hypothesis, conceivably explaining why highly pure mammalian spermatogonia have not developed into meiosis, much less through meiosis without somatic cells...
2017: Methods in Molecular Biology
Aileen R Helsel, Jon M Oatley
In mammals, the activities of spermatogonial stem cells (SSCs) provide the foundation for continual spermatogenesis throughout a male's reproductive lifetime. At present, the defining characteristics of SSCs and mechanisms controlling their fate decisions are not well understood. Transplantation is a definitive functional measure of stem cell capacity for male germ cells that can be used as an assay to provide an unequivocal quantification of the SSC content in an experimental cell population. Here, we discuss the procedure for mice and provide protocols for preparing donor germ cell suspensions from testes directly or primary cultures of spermatogonia for transplantation, enriching for SSCs, preparing recipient males, microinjection into recipient testes, and considerations for experimental design...
2017: Methods in Molecular Biology
Gina M Aloisio, Ileana Cuevas, Yuji Nakada, Christopher G Peña, Diego H Castrillon
The precise identity of spermatogonial stem cells-the germline stem cell of the adult testis-remains a controversial topic. Technical limitations have included the lack of specific markers and methods for lineage tracing of Asingle spermatogonia and their subsets. Immunolocalization of proteins in tissue sections has been a standard tool for the in situ identification and visualization of rare cellular subsets. However, these studies are limited by the need for faithful and reliable protein markers to define these cell types, as well as the availability of specific antibodies to these markers...
2017: Methods in Molecular Biology
Kazadi Mutoji, Anukriti Singh, Thu Nguyen, Heidi Gildersleeve, Amy V Kaucher, Melissa J Oatley, Jon M Oatley, Ellen K Velte, Chris B Geyer, Keren Cheng, John R McCarrey, Brian P Hermann
Precise separation of spermatogonial stem cells (SSCs) from progenitor spermatogonia that lack stem cell activity and are committed to differentiation remains a challenge. To distinguish between these spermatogonial subtypes, we identified genes that exhibited bimodal mRNA levels at the single cell level among undifferentiated spermatogonia from the Postnatal Day 6 (P6) mouse testis, including Tspan8, Epha2 and Pvr, each of which encode cell-surface proteins useful for cell selection. Transplantation studies provided definitive evidence that a TSPAN8-high subpopulation is enriched for SSCs...
October 12, 2016: Biology of Reproduction
Charles Teta, Yogeshkumar S Naik
Industrial and municipal effluents regularly pollute water bodies and cause various toxic effects to aquatic life. Because of the diverse nature of industrial processes and domestic products, urban effluents are often tainted with various anthropogenic endocrine disrupting chemicals that may interfere with the reproductive physiology of aquatic fauna. In this study, we tested effluents from the City of Bulawayo for the presence of estrogenic endocrine disrupting chemicals and their effects on fish gonads and fecundity...
October 8, 2016: Chemosphere
Kate A Redgrove, Ilana R Bernstein, Victoria J Pye, Bettina P Mihalas, Jessie M Sutherland, Brett Nixon, Adam McCluskey, Phillip J Robinson, Janet E Holt, Eileen A McLaughlin
The dynamin family of proteins play important regulatory roles in membrane remodelling and endocytosis, especially within brain and neuronal tissues. In the context of reproduction, dynamin 1 (DNM1) and dynamin 2 (DNM2) have recently been shown to act as key mediators of sperm acrosome formation and function. However, little is known about the roles that these proteins play in the developing testicular germ cells. In this study, we employed a DNM2 germ cell-specific knockout model to investigate the role of DNM2 in spermatogenesis...
October 11, 2016: Scientific Reports
Christiane Pleuger, Daniela Fietz, Katja Hartmann, Wolfgang Weidner, Sabine Kliesch, Moira K O'Bryan, Adriaan Dorresteijn, Martin Bergmann
OBJECTIVE: To define the stage-by-stage expression of KATNB1 during human spermatogenesis. DESIGN: Gene expression analysis, histologic and immunohistochemical evaluation. SETTING: University research laboratories and andrological clinic. PATIENT(S): Eighty human testicular biopsy samples: 43 showing normal spermatogenesis, 9 with maturation arrest at level of spermatocytes, 8 with maturation arrest at level of spermatogonia, and 20 with a Sertoli cell only syndrome...
October 4, 2016: Fertility and Sterility
Ieva Masliukaite, Julie M Hagen, Kirsi Jahnukainen, Jan-Bernd Stukenborg, Sjoerd Repping, Fulco van der Veen, Madelon van Wely, Ans M M van Pelt
OBJECTIVE: To collect published data on spermatogonial quantity in the testes of healthy children and calculate the reference values of spermatogonial quantities throughout prepuberty. DESIGN: Systematic literature search in PubMed and EMBASE focusing on the number of spermatogonia per transverse tubular cross section (S/T) and spermatogonial density per cubic centimeter (cm(3)) of testicular volume (S/V) throughout prepuberty. SETTING: None...
October 4, 2016: Fertility and Sterility
T G Borovskaya, A M Dygai, Yu A Shchemerova, S I Kamalova, V A Mashanova, A V Vychuzhanina, M E Poluektova, P G Madonov, D N Kinsht, V E Goldberg
Effectiveness of the granulocyte colony-stimulating factor immobilized by using electronbeam synthesis nanotechnology was investigated on the model of experimental testicular failure caused by the toxic effect on stem spermatogonia. Administration of the drug to experimental paclitaxel-treated animals increased the number of sources of the proliferative pool of spermatogenesis and its productivity. The effectiveness of immobilized granulocyte colony-stimulating factor was based on its ability to stimulate reparative regeneration of the spermatogenic tissue, which manifested in a decrease in spermatogenic layer maturity and increase in the number of microenvironment cells...
September 2016: Bulletin of Experimental Biology and Medicine
R Kumar Pramod, Bo Ram Lee, Young Min Kim, Hong Jo Lee, Young Hyun Park, Tamao Ono, Jeong Mook Lim, Jae Yong Han
We reported previously testis-mediated germline chimera production and characterization of germline stem cell-like cells from chicken testes. The present study aimed to establish an in vitro system for culture of quail spermatogonia stem cells (SSCs) for practical applications in germline preservation and transgenesis. Testicular cells from juvenile (4 weeks old) or adult (8 weeks old) quail testis were isolated using sequential enzymatic digestion. The percentages of viability of isolated testicular cells were 91...
October 5, 2016: Stem Cells and Development
Weiyi Zhang, Yuanyuan Shao, Yinghe Qin, Yingjie Wu
Heat shock factors (HSFs) are critical regulators of spermatogenesis. However, heat shock responses, the associated components and the underlying functional mechanisms remain to be elucidated. Here, we characterize the expression pattern of HSFY, a member of the HSF family in the testis and epididymis. Its expression in testis and epididymis was initially identified by western blots. Immunofluorescence staining demonstrated that HSFY was confined to the cytoplasm of late spermatocytes and spermatids in adult testes, gonocytes in newborn testes and undifferentiated spermatogonia in 7 days post-parturition testes...
September 30, 2016: Cell and Tissue Research
Diego Safian, Roberto D Morais, Jan Bogerd, Rüdiger W Schulz
Insulin-like growth factor (Igf) binding proteins (Igfbps) modulate the availability of Igfs for their cognate receptors. In zebrafish testes, Igf3 promotes the proliferation and differentiation of type A undifferentiated (Aund) spermatogonia, and igf3 expression is strongly elevated by Fsh, but also responds to T3. Here, we report the effects of Fsh and T3 on igfbp transcript levels in adult zebrafish testis. We then examined T3 and Fsh effects on zebrafish spermatogenesis and explored the relevance of Igfbps in modulating these T3 or Fsh effects, using a primary tissue culture system for adult zebrafish testis...
September 30, 2016: Endocrinology
T Mirzapour, T A B Tengku Ibrahim, M Movahedin, M R Nowroozi
Destruction of spermatogonial stem cells (SSCs) along the chemotherapy and radiotherapy is one of the side effects of cancer treatments that lead to infertility. In vitro propagation of hSSCs is necessary to obtain an adequate number of cells for successful transplantation. In this study, hSSCs were isolated from testis biopsies of the patients with maturation arrest and proliferated in DMEM in the presence of LIF and bFGF for 5 weeks. The various types of human spermatogonia were identified in culture system and compared with testis tissue using morphological criteria at the ultrastructural level...
September 29, 2016: Andrologia
Hye-Won Song, Anilkumar Bettegowda, Blue B Lake, Adrienne H Zhao, David Skarbrevik, Eric Babajanian, Meena Sukhwani, Eleen Y Shum, Mimi H Phan, Terra-Dawn M Plank, Marcy E Richardson, Madhuvanthi Ramaiah, Vaishnavi Sridhar, Dirk G de Rooij, Kyle E Orwig, Kun Zhang, Miles F Wilkinson
The developmental origins of most adult stem cells are poorly understood. Here, we report the identification of a transcription factor-RHOX10-critical for the initial establishment of spermatogonial stem cells (SSCs). Conditional loss of the entire 33-gene X-linked homeobox gene cluster that includes Rhox10 causes progressive spermatogenic decline, a phenotype indistinguishable from that caused by loss of only Rhox10. We demonstrate that this phenotype results from dramatically reduced SSC generation. By using a battery of approaches, including single-cell-RNA sequencing (scRNA-seq) analysis, we show that Rhox10 drives SSC generation by promoting pro-spermatogonia differentiation...
September 27, 2016: Cell Reports
L Dumont, A Oblette, C Rondanino, F Jumeau, A Bironneau, D Liot, V Duchesne, J Wils, N Rives
STUDY QUESTION: Does vitamin A (retinol, Rol) prevent round spermatid nuclear damage and increase the production of motile sperm during in vitro maturation of vitrified pre-pubertal mouse testicular tissue? SUMMARY ANSWER: The supplementation of an in vitro culture of ~0.75 mm(3) testicular explants from pre-pubertal mice with Rol enhances spermatogenesis progression during the first spermatogenic wave. WHAT IS KNOWN ALREADY: The production of functional spermatozoa in vitro has only been achieved in the mouse model and remains a rare event...
September 25, 2016: Molecular Human Reproduction
Ryusuke Nakajima, Hideyuki Okano, Toshiaki Noce
Jmjd1C is one of the Jmjd1 family genes that encode putative demethylases against histone H3K9 and non-histone proteins and has been proven to play an indispensable role in mouse spermatogenesis. Here, we analyzed a newly-bred transgenic mouse strain carrying a Jmjd1C loss-of-function allele in which a β-geo cassette was integrated into the intron of the Jmjd1C locus. Jmjd1C gene-trap homozygous testes exhibited malformations in postmeiotic processes and a deficiency in the long-term maintenance of undifferentiated spermatogonia...
2016: PloS One
Maryam Nazm Bojnordi, Hossein Azizi, Thomas Skutella, Mansoureh Movahedin, Fereshteh Pourabdolhossein, Amir Shojaei, Hatef Ghasemi Hamidabadi
Transplantation of embryonic stem cells (ESCs) is a promising therapeutic approach for the treatment of neurodegenerative diseases. However, ESCs are not usable clinically due to immunological and ethical limitations. The identification of an alternative safe cell source opens novel options via autologous transplantation in neuro-regeneration circumventing these problems. Here, we examined the neurogenic capacity of embryonic stem-like cells (ES-like cells) derived from the testis using neural growth factor inducers and utilized them to generate functional mature neurons...
September 19, 2016: Molecular Neurobiology
Dina Cortes, Rune Holt, Victoria Elizabeth de Knegt
A normal functioning hypothalamic-pituitary-testicular axis is required for normal testicular descent. The percentage of cases that result from a disturbance in this axis remains controversial. Much has yet to be learnt about cryptorchidism, but is seems that the existence of A dark spermatogonia (Ad spermatogonia) is essential for later fertility. Bilateral cryptorchid patients have a high risk of later infertility, even though they undergo early surgery for cryptorchidism. It is possible today to distinguish-to a certain extent-between three different groups of cryptorchid patients based on testicular histology, gonadotropins, and inhibin B at the time of early surgery: Group 1, patients suspected of prepubertal transient hypothalamic-pituitary-testicular hypofunction and a high risk of later infertility; Group 2, patients with hypergonadotropic hypogonadism and a primary testicular dysfunction; and Group 3, patients with normal histology and normal serum levels of inhibin B and gonadotropins at the time of early surgery and a low risk of later infertility...
October 2016: European Journal of Pediatric Surgery
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