Localization and quantitative expression of mRNAs encoding the testis-specific histone TH2B, the phosphoprotein p19, the transition proteins 1 and 2 during pubertal development and throughout the spermatogenic cycle of the rat

Marret, Cécile; Avallet, O.; Perrard-Sapori, Marie-Hélène; Durand, Philippe

doi:10.1002/(sici)1098-2795(199809)51:1<22::aid-mrd3>3.0.co;2-y

Cited by 33 publications

(31 citation statements)

References 39 publications

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“…8). As expected [13], very low quantities of both TP1 and TP2 mRNAs could be measured during the first week of the experiments, when no postmeiotic cell was present. However, an exponential-like increase in the amounts of the mRNAs encoding for TP1 (r ϭ 0.762, P Ͻ 0.01) or TP2 (r ϭ 0.693, P Ͻ 0.01) occurred from the time when 1C cells appeared in the culture until the end of the experiment.…”

Section: Resultssupporting

confidence: 77%

“…However, it seems unlikely that the increase in TP1 and TP2 mRNA levels can be explained solely by the differentiation of a small number of young spermatids that express these mRNA, at a high level. We have shown previously [13] that TP1 and TP2 mRNA levels increase greatly only from steps 6 or 7 of spermiogenesis and under our culture conditions, round spermatids beyond stages III-IV were never observed (unpublished results).…”

Section: Discussionsupporting

confidence: 68%

“…Very importantly, these spermatids formed in vitro were able to transcribe postmeiotic-specific genes [13]. Moreover, it must be noted that the levels of the mRNA encoding for TP1 or TP2 increased until the end of the culture even if the number of 1C cells in the culture remained constant or even decreased.…”

Section: Discussionmentioning

confidence: 98%

“…Moreover, it must be noted that the levels of the mRNA encoding for TP1 or TP2 increased until the end of the culture even if the number of 1C cells in the culture remained constant or even decreased. This can be easily explained by the fact that the transcription of TP1 and TP2 increases when round spermatids mature [13] and that the very early steps of spermiogenesis can occur in vitro. However, it seems unlikely that the increase in TP1 and TP2 mRNA levels can be explained solely by the differentiation of a small number of young spermatids that express these mRNA, at a high level.…”

Section: Discussionmentioning

confidence: 99%

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The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

Staub

Hue

Nicolle

et al. 2000

Experimental Cell Research

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The aim of the present study was to assess whether the whole meiotic process of spermatogenic cells is able to take place in vitro. Fragments of seminiferous tubules from 20-to 22-or 28-day-old rats were seeded in medium containing 0.2% fetal calf serum in bicameral chambers and then cultured for 4 weeks in a chemically defined medium. The differentiation of meiotic germinal cells was followed by four criteria: (i) ultramicroscopic examination of the different types of germ cells present in the cell layer throughout the culture period; (ii) determination of the changes in DNA content per nucleus of the cell population seeded with time in culture; (iii) assessment of the ability of germinal cells to transcribe genes expressed after completion of meiosis; and (iv) monitoring the fate of BrdU-labeled leptotene spermatocytes. The ultrastructural study showed that the overall organization of the cells in the culture well recalls that of the seminiferous epithelium throughout the culture period. Moreover the identification of young round spermatids 21 days after seeding suggested that these spermatids had been formed very recently in culture. Determination of DNA content per nucleus showed that a 1C cell population could be observed after several days of cultures reaching 6 to 10% of total cells. An exponential-like increase in the amounts of the mRNAs encoding for TP1 or TP2 occurred from the time when 1C cells appeared in the culture until the end of the experiment. Finally, BrdU-labeled leptotene spermatocytes differentiated into pachytene spermatocytes and then into secondary spermatocytes, and BdrU-labeled round spermatids were observed from Day 21 of culture onward. Taken together these results indicate that the whole meiotic process from leptotene spermatocyte to round spermatid can indeed occur in vitro under the present culture conditions.

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Section: Resultssupporting

confidence: 77%

Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

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The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

Staub

Hue

Nicolle

et al. 2000

Experimental Cell Research

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“…During post-natal development, the timing of the appearance of the germ cells was concordant with the description given by Clermont and Perrey (1957) and Marret et al (1998). Briefly, the most advanced germ cells were spermatogonia at 10 days, spermatocytes at 20 days, round spermatids at 30 days, elongated spermatids at 40 days and spermatozoa at 60 days.…”

Section: Validation Of Clusterin Mrna Level As a Stable Marker Of Thesupporting

confidence: 76%

Quantification of stem cell factor mRNA levels in the rat testis: usefulness of clusterin mRNA as a marker of the amount of mRNA of sertoli cell origin in post pubertal rats

Plotton¹,

Sanchez²,

Perrard³

et al. 2005

Journal of Endocrinology

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Spermatogenesis is a complex cellular process regulated by gonadotrophins and local cell-cell interactions. Stem cell factor (SCF) is one of the paracrine factors, produced by the Sertoli cells, involved in the local regulation of spermatogenesis. Measurement of its testicular level is important for addressing its role in testis physiopathology. However, the relative cell composition of experimental and pathological testis samples may lead to misinterpretation in relating SCF mRNA levels to the amount of RNA extracted from the whole tissue sample. Taking into account the relative RNA content of Sertoli cell origin should provide more significant data. In the present study, three sets of experiments were intended for modifying the proportion of RNA of Sertoli cell origin in RNA extracted from whole testis tissue samples: during postnatal development; following methoxy-acetic acid (MAA) administration; and after injecting a long-acting gonadotrophin-releasing hormone agonist (GnRHa). In a first step, we demonstrated clusterin mRNA level stability in purified Sertoli cell preparations between 20 days and adulthood, and following MAA or GnRHa treatment. In a second step, we used a competitive RT-PCR assay to measure SCF and clusterin mRNA levels and expressed the amount of SCF mRNA relative to the amount of clusterin mRNA under the above experimental conditions. The SCF/clusterin mRNA level ratio was found to remain roughly stable from 20 days post-partum to adulthood; i.e. during the development of spermatogenesis. MAA administration led to an overall increase in the SCF/clusterin mRNA level ratio between 7 and 14 days after administration, consistent with the replenishment of the testis with pachytene spermatocytes and round spermatids. Conversely, after long-acting GnRHa injection, the SCF/clusterin mRNA level ratio decreased only slightly from day 21 onward. Hence, the present studies indicate that, under physiopathological conditions, the amount of clusterin mRNA is a good marker of the amount of RNA of Sertoli cell origin in testis samples at day 20 or later; different experimental alterations of spermatogenesis are associated with different patterns of SCF mRNA levels; the relationship between FSH and SCF in vivo is not as simple as that described in vitro.

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Cellular and subcellular localization of stathmin during oocyte and preimplantation embryo development

et al. 1999

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Stathmin is a 19 kDa cytosolic phosphoprotein, proposed to act as a relay integrating diverse intracellular signaling pathways involved in regulation of cell proliferation, differentiation, and function. To gain further information about its significance during early development, we analyzed stathmin expression and subcellular localization in mouse oocytes and preimplantation embryos. RT‐PCR analysis revealed a low expression of stathmin mRNA in unfertilized oocytes and a higher expression at the blastocyst stage. A fine cytoplasmic punctuate fluorescent immunoreactive stathmin pattern was detected in the oocyte, while it evolved toward an increasingly speckled pattern in the two‐cell and later four‐ to eight‐cell embryo, with even larger speckles at the morula stage. In blastocysts, stathmin immunoreactivity was fine and intense in inner cell mass cells, whereas it was low and variable in trophectodermal cells. Electron microscopic analysis allowed visualization with more detail of two types of stathmin immunolocalization: small clusters in the cytoplasm of oocytes and blastocyst cells, together with loosely arranged clusters around the outer membrane of cytoplasmic vesicles, corresponding to the immunofluorescent speckles in embryos until the morula stage. In conclusion, it appears from our results that maternal stathmin is accumulated in the oocyte and is relocalized within the oocyte and early preimplantation embryonic cell cytoplasm to interact with specific cytoplasmic membrane formations. Probably newly synthesized, embryonic stathmin is expressed in the blastocyst, where it is localized more uniformly in the cytoplasm mostly of inner cell mass (ICM) cells. These expression and localization patterns are probably related to the particular roles of stathmin at the successive steps of oocyte maturation and early embryonic development. They further support the proposed physiologic importance of stathmin in essential biologic regulation. Mol. Reprod. Dev. 53:306–317, 1999. © 1999 Wiley‐Liss, Inc.

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Localization and quantitative expression of mRNAs encoding the testis-specific histone TH2B, the phosphoprotein p19, the transition proteins 1 and 2 during pubertal development and throughout the spermatogenic cycle of the rat

Cited by 33 publications

References 39 publications

The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

Quantification of stem cell factor mRNA levels in the rat testis: usefulness of clusterin mRNA as a marker of the amount of mRNA of sertoli cell origin in post pubertal rats

Cellular and subcellular localization of stathmin during oocyte and preimplantation embryo development

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