Quantitative study of the cell population of the seminiferous tubules in immature rats

Clermont, Y.; Perey, Bernard

doi:10.1002/aja.1001000205

Cited by 702 publications

(399 citation statements)

References 19 publications

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“…The 4C to vimentin‐positive cell ratio obtained in this study from the adult rat testis was 7.6, indicating that a portion of the adult Sertoli cells are lost during the sample preparation because of incomplete digestion as expected. A similar phenomenon was observed in the 5‐ and 10‐day‐old testes when compared to reference values from the literature (Clermont & Perey, 1957). Age, species, and testicular fibrosis can contribute to an incomplete digestion and care should be taken when applying this method to experimental groups with very different cell composition.…”

Section: Discussionsupporting

confidence: 88%

“…The ratios of the different cell populations during development mirrored the dynamics assessed previously using histomorphometric methods (Clermont & Perey, 1957; Roosen‐Runge & Leik, 1968). With even PI alone, the emergence of the meiotic cells from post‐natal day (PND)16 onwards and appearance of the first haploid germ cells at PND24 in the rat testis could be assessed.…”

Section: Discussionsupporting

confidence: 70%

“…In the 16‐day‐old rat testis, there are no spermatids and as expected there were no γH2AX‐positive 1C cells either. A larger proportion of the 2C cells were positive for γH2AX in the 16‐day‐old than in the adult testis which corresponded to the amplification of the intermediate and B spermatogonia at this time‐point (Clermont & Perey, 1957). These results show that the γH2AX assay functions in the rat testis and it could be applied in the future to detect the different testicular germ cell populations.…”

Section: Discussionmentioning

confidence: 99%

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A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

et al. 2015

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SummaryAccurate analysis and quantification of different testicular cell populations are of central importance in studies of male reproductive biology. The traditional histomorphometric and immunohistochemical methods remain the gold standard in studying the complex dynamics of the testicular tissue. Through past years advances have been made in the application of flow cytometry for the rapid analysis of testicular cell populations. Detection of DNA content and of surface antigens and fluorescent reporters have been widely used to analyze and sort cells. Detection of intracellular antigens can broaden the possibilities of applying flow cytometry in studies of male reproduction. Here, we report a detailed protocol for the preparation of rat testicular tissue for detection of intracellular antigens by flow cytometry, and a pipeline for subsequent data analysis and troubleshooting. Rat testicular ontogenesis was chosen as the experimental model to validate the performance of the assay using vimentin and γH2AX as intracellular markers for the somatic and spermatogenic cells, respectively. The results show that the assay is reproducible and recapitulates the rat testis ontogenesis.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionsupporting

confidence: 70%

Section: Discussionmentioning

confidence: 99%

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A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

et al. 2015

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“…Their neonatal differentiation is an essential process for the development and maturation of mammalian testes and requires their proliferation, relocation to the basement membrane of seminiferous epithelium and apoptosis (Clermont and Perey, 1957;Magre and Jost, 1980;McGuinness and Orth, 1992a,b;Brennan and Capel, 2004;Sakai et al, 2004). Two phases of Gn proliferation occur in some rodents such as rat, the first in the embryonic period and the second at 4 days of age (Miething, 1992;Wang et al, 1998;Boulogne et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

“…Two phases of Gn proliferation occur in some rodents such as rat, the first in the embryonic period and the second at 4 days of age (Miething, 1992;Wang et al, 1998;Boulogne et al, 1999). Between these mitotic phases Gn remains quiescent (Clermont and Perey, 1957;Magre and Jost, 1980;McGuinness and Orth, 1992b). In rats and hamsters, the periods of Gn mitotic activity coincide with the highest apoptotic rate (Miething, 1992;Boulogne et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Neonatal Gonocyte Differentiation in Mongolian GerbilMeriones unguiculatusInvolves Asynchronous Maturation of Seminiferous Cords and Rapid Formation of Transitional Cell Stage

Pinto

Botta

Taboga

et al. 2010

The Anatomical Record

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This study describes the neonatal differentiation of the Mongolian gerbil gonocytes, focusing on the relationship between its relocation to the basement membrane, apoptosis and postrelocation changes and also the distribution of androgen receptors (AR). Testes of gerbils from 1 to 35 days of age (d) were examined by high resolution light microscopy and immunocytochemistry for proteins PCNA, VASA, and AR as well as by the TUNEL method. Gonocytes were quantified according to degree of relocation into nonrelocated, relocating and relocated. Most of them were found in the center of seminiferous cords at 1 d but a small number of relocating and relocated gonocytes were already visible in the first postnatal day. After relocation, gonocytes change phenotypically to a transitional stage designated herein prospermatogonia. Both gonocyte relocation and transformation into spermatogonial lineage occur asynchronously in the seminiferous cords, mainly after 7 d. Gonocyte proliferation began before but peak after their relocation to basement membrane at the prospermatogonia stage. Higher levels of gonocyte apoptosis were found at 7 d and 21 d. From this time onward gonocytes were not found. Gonocytes and prospermatogonia showed high amounts of AR in their cytoplasm contrary to spermatogonial subtypes, indicating a possible AR inactivation in these cells. In conclusion, the process of gonocyte relocation in the gerbil extends until the second postnatal week, leads to their rapid differentiation into prospermatogonia and occurs simultaneously with the loss of androgen sensitivity. Differently from other laboratory rodents, the events regarding gonocyte maturation in the gerbil last longer and occur asynchronously in seminiferous cords. Anat Rec,

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Some remarks on the female and male Keimbahn in the light of evolution and history

Hilscher

1999

J. Exp. Zool.

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From the existence of two types of cells for reproduction—the female and male germ cells (GCs)—and by recombination of the genome, evolution proceeded dramatically. Unicellular and multicellular plants frequently are characterized by a sequence of haploid and diploid phases, or generations, with gametes and spores as reproductive cells. Isogamy, anisogamy, and oogamy can be distinguished depending on the GCs that correspond, differ in size, or impose as egg cell and sperm cell. In protozoans, too, species are found in which GCs differ clearly from each other. In the female lineage of angiosperms, a “Keimbahn chain” consisting of five successive germ line cells can be observed. Oogenesis and spermatogenesis are complete in coelenterates and similar in mammals. However, the controlling mechanisms are by far more complex in the latter. This means that the balance of hormonal and vegetative nervous influences (stimulation, inhibition) on gametogenesis is not primarily orientated on the germ line cells themselves, but mostly on the structural and functional situation of the gonads and the individual carriers. This becomes particularly evident in insects, where gametogenesis, on the one side, depends on the development of the rest of the organism but on the other side represents an independent developmental process. The point at which germ line cells and somatic cells separate correlates more or less with the degree of phylogenetic development. In worms, insects, and up to the anurans, a part of the cytoplasm, the so‐called germ plasma, is separated for the development of GCs during oogenesis (preformistic development). However, in urodeles, reptiles, birds, and mammals, GCs and somatic cells cannot be distinguished before gastrulation (epigenetic development). In various species (e.g., in some oligochaetes and snails), there exist “double spermatogenic lines.” In mammals (probably in other vertebrates and perhaps in various phyla of animals, too), the female Keimbahn is provided with only one proliferation system. The male gametogenesis is equipped with two systems: the first corresponds to the female germ line, the second is responsible for the immense number of gametes produced in the mature testes. In mammals the message to become male lies on the Y‐chromosome (on its short arm in man and mouse) and was identified as the gene SRY in human and Sry in mouse. The fertility genes that are responsible for an uninterrupted spermatogenesis, up to fertilizing spermatozoa, are sitting on the long arm of the human Y‐chromosome. J. Exp. Zool. (Mol. Dev. Evol.) 285:197–214, 1999. © 1999 Wiley‐Liss, Inc.

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Quantitative study of the cell population of the seminiferous tubules in immature rats

Cited by 702 publications

References 19 publications

A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

Neonatal Gonocyte Differentiation in Mongolian GerbilMeriones unguiculatusInvolves Asynchronous Maturation of Seminiferous Cords and Rapid Formation of Transitional Cell Stage

Some remarks on the female and male Keimbahn in the light of evolution and history

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