A reassessment of Y chromosomal behaviour in germ cells and Sertoli cells of the mouse as revealed by in situ hybridisation

Hypothalamic Gonadotropin Releasing Hormone (GnRH),viaGnRH receptor (GnRHR), is the main actor in the control of reproduction, in that it induces the biosynthesis and the release of pituitary gonadotropins, which in turn promote steroidogenesis and gametogenesis in both sexes. Extrabrain functions of GnRH have been extensively described in the past decades and, in males, local GnRH activity promotes the progression of spermatogenesis and sperm functions at several levels. The canonical localization ofGnrh1andGnrhr1mRNA is Sertoli and Leydig cells, respectively, but ligand and receptor are also expressed in germ cells. Here, we analysed the expression rate ofGnrh1andGnrhr1in rat testis (180 days old) by quantitative real-time PCR (qPCR) and byin situhybridization we localizedGnrh1andGnrhr1mRNA in different spermatogenic cells of adult animals. Our data confirm the testicular expression ofGnrh1and ofGnrhr1in somatic cells and provide evidence that their expression in the germinal compartment is restricted to haploid cells. In addition, not only Sertoli cells connected to spermatids in the last steps of maturation but also Leydig and peritubular myoid cells expressGnrh1.

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XY follicle cells in the ovaries of XO/XY and XO/XY/XYY mosaic mice

Development ◽

10.1242/dev.111.4.1017 ◽

1991 ◽

Vol 111 (4) ◽

pp. 1017-1019 ◽

Cited By ~ 2

Author(s):

S.J. Palmer ◽

P.S. Burgoyne

Keyword(s):

Sertoli Cells ◽

Ovarian Development ◽

In Situ Hybridisation ◽

Cell Lineage ◽

Supporting Cell ◽

Follicle Cells ◽

Cell Lineages ◽

Determination Process ◽

Testis Determination

XO/XY and XO/XY/XYY mosaic hermaphrodites were generated from crosses involving BALB/cWt males. The distribution of Y-bearing cells in the gonads of these mice was studied by in situ hybridisation using the Y-specific probe pY353B. XY cells were found to contribute to all cell lineages of the ovary including follicle cells. The proportion of XY follicle cells was not significantly different from the XY contribution to other gonadal or non-gonadal cell lineages. However, this proportion was consistently low, all the hermaphrodites having a low XY contribution to the animal as a whole. Because the XO- and Y-bearing cell lineages are developmentally balanced, the XY follicle cells cannot have formed as a result of a ‘mismatch’ in which the Y-directed testis determination process is pre-empted by an early acting programme of ovarian development. These results are discussed with respect to the hypothesis that Tdy acts in the supporting cell lineage, the lineage from which Sertoli cells and follicle cells are believed to be derived.

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218. Expression of Wsb2 in the mouse testis

Reproduction Fertility and Development ◽

10.1071/srb05abs218 ◽

2005 ◽

Vol 17 (9) ◽

pp. 84

Author(s):

M. Sarraj ◽

P. J. McClive ◽

K. L. Loveland ◽

A. H. Sinclair

Keyword(s):

Sertoli Cells ◽

Leydig Cells ◽

Adult Mouse ◽

In Situ Hybridisation ◽

Mouse Testis ◽

Male Gonad ◽

Whole Mount ◽

Mantle Layer ◽

Pachytene Spermatocytes

We present a detailed study on the expression pattern of Wsb2 in the mouse foetal and adult gonad. Wsb2 expression was analysed during mouse embryogenesis by whole-mount, section in situ hybridisation and immunohistochemistry. Wsb2 was found to be expressed in the developing mouse gonads from 11.5 dpc to 16.5 dpc. Expression is initially equal in both sexes from 10.5 dpc until 12.0 dpc, then it persists in the male gonad. Wsb2 expression was confined to the cords in both Sertoli cell and germ cells. Other sites of Wsb2 embryonic expression were the somites, dorsal root ganglia and the lateral mantle layer of the neural tube. mRNA encoding Wsb2 and Wsb2 protein has been detected in the newborn testis in both gonocytes and Sertoli cells. Wsb2 mRNA in the adult mouse testis was observed in Sertoli cells, spermatogonia, spermatocytes and the corresponding Wsb2 protein expression was in pachytene spermatocytes, round and elongated spermatids, Sertoli cells and Leydig cells. The differential expression of Wsb2 in male versus female embryonic gonads suggests it may play a role in mammalian sex determination during embryonic development and its expression in the first wave of spermatogenesis and in the adult suggests a later role in spermatogenesis.

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Investigation of the potential role of the germ cell complement in control of the expression of transferrin mRNA in the prepubertal and adult rat testis

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0190067 ◽

1997 ◽

Vol 19 (1) ◽

pp. 67-77 ◽

Cited By ~ 11

Author(s):

S M Maguire ◽

M R Millar ◽

R M Sharpe ◽

J Gaughan ◽

P T K Saunders

Keyword(s):

Germ Cell ◽

Mrna Expression ◽

Germ Cells ◽

Sertoli Cells ◽

Direct Access ◽

Adult Rats ◽

Rat Testis ◽

Adult Rat

ABSTRACT Iron is required for the normal development of germ cells during spermatogenesis. Because these cells have no direct access to systemic iron, there exists a shuttle system involving production and secretion of the iron-transporting protein transferrin by the Sertoli cells. Previous reports using cultures of immature Sertoli cells exposed to adult germ cells, or in vivo studies involving germ cell-depleted adult rat testes, concluded that production of transferrin by Sertoli cells is modulated by germ cell complement. In the present study we have used in situ hybridisation with cRNA probes directed against the 5′ and 3′ ends of transferrin mRNA to examine the pattern of expression of transferrin in the immature and adult rat testis. Adult rats were treated with ethane dimethane sulphonate or methoxyacetic acid (MAA) to manipulate their testosterone levels or germ cell complement respectively. Initial findings obtained using the 3′ probe showed a decrease in transferrin mRNA associated with round spermatid depletion. However, these data were not confirmed by in situ hybridisation when the 5′ probe was used. The specificity of the probes was examined using Northern blotting and the 3′ probe was found to hybridise to the germ cell transcript for hemiferrin even under conditions of high stringency. Examination of immature and pubertal rat testes by in situ hybridisation using the 5′ transferrin-specific probe found that as early as 14 days of age the level of expression of transferrin mRNA was clearly different between tubules, and the mRNA appeared to be expressed in Leydig cells on and after day 31. In the adult rat testis, maximal expression of transferrin mRNA was found at stages VIII-XIV, calling into question the interpretation of the results of some previous studies showing expression of transferrin mRNA at all stages of the spermatogenic cycle. This stage-specific pattern of expression was not altered by acute germ cell depletion using MAA. However, Northern blot analysis showed a statistically significant increase in transferrin mRNA expression at 7 days after MAA treatment when pachytene spermatocytes were depleted from tubules at all stages of the spermatogenic cycle at which transferrin is normally expressed. In conclusion, we found that transferrin mRNA expression was not modulated by round spermatids as has been reported previously but that meiotic germ cells may influence expression of transferrin at specific stages of the spermatogenic cycle.

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Combined use of proacrosion immunocytochemistry and autosomal DNA in situ hybridisation for evaluvation of human ejaculated germ cells

Zygote ◽

10.1017/s0967199400003233 ◽

1996 ◽

Vol 4 (04) ◽

pp. 279-283 ◽

Cited By ~ 9

Author(s):

Carmen Mendoza ◽

Moncef Benkhalifa ◽

Paul Cohen-Bacrie ◽

André Hazout ◽

Yves Ménézo ◽

...

Keyword(s):

Germ Cells ◽

In Situ Hybridisation ◽

Round Spermatid ◽

Testicular Tissue ◽

Obstructive Azoospermia ◽

Structural Pattern ◽

Combined Use ◽

Cytoplasmic Maturation ◽

Pre Treatment

SummaryThe recently reported human pregnancies and births after fertilising oocytes with round spermatids recovered from the ejaculate of men with non-obstructive azoospermia have underscored the need for a more accurate evaluation of the nuclear and cytoplasmic maturation status of ejaculated germ cells. In this study we describe our first experience with a method combining the immunocytochemical visualisation of proacrosin with autosomal DNA fluorescencein situhybridisation (FISH) to assess ejaculated germ cells from patients with a spermiogenesis defect. The proacrosin immunoreactivity, analysed with the use of the monoclonal antibody 4D4, has been detected in cells of round spermatid size presenting a haploid FISH figure as well as in larger cells whose ploidy corresponds to primary and secondary spermatocytes. These observations are in agreement with previously published results obtained, with the use of the same antibody, by immunocytochemical analysis of histological sections of testicular tissue. All the cells of round spermatid size possessing proacrosin immunoreactivity were found to be haploid by FISH. On the other hand, some of the haploid cells of round spermatid size did not possess proacrosin immunoreactivity. The structural pattern of proacrosin immunoreactivity was highly variable both in spermatids and in younger spermatogenic cells. These data show that cell size is the main criterion to be used for the identification of ejaculated round spermatids, whereas the presence of the developing acrosome represents only an auxiliary criterion. The scoring of acrosomal development in ejaculated spermatids may be useful as part of pre-treatment diagnosis before the inclusion of infertile couples in a spermatid conception programme.

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In situ analysis of fetal, prepuberal and adult XX—XY chimaeric mouse testes: Sertoli cells are predominantly, but not exclusively, XY

Development ◽

10.1242/dev.112.1.265 ◽

1991 ◽

Vol 112 (1) ◽

pp. 265-268 ◽

Cited By ~ 25

Author(s):

S.J. Palmer ◽

P.S. Burgoyne

Keyword(s):

Somatic Cell ◽

Sertoli Cell ◽

Sertoli Cells ◽

In Situ Hybridisation ◽

Age Groups ◽

Cell Types ◽

Beta Globin ◽

Cell Type ◽

Cell Lineages

The testes of fetal, prepuberal and adult XX—XY chimaeras were examined using in situ hybridisation to identify the beta-globin transgenic marker contained in one component of each chimaera. This enabled the proportion of XX and XY cells contributing to the major cell lineages of the testis to be estimated from sectioned and air-dried material. A few XX Sertoli cells were found in all three age groups, but the XX contribution was always much lower than in other somatic cell types. Significantly, in fetal XX—XY testes, Sertoli cells were the only cell type to show a bias in favour of the XY component. This strengthens the view that Tdy acts solely in the lineage that gives rise to Sertoli cells. However, the finding of some fetal XX Sertoli cells means that one of the steps in the Tdy-initiated process of Sertoli cell determination is capable of locally recruiting XX cells.

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Freeze-etch observations on the tight junctions of sertoli cells grown in organ culture with FSH

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010008256x ◽

1978 ◽

Vol 36 (2) ◽

pp. 340-341

Author(s):

Rita Meyer ◽

Zoltan Posalaky ◽

Dennis Mcginley

Keyword(s):

Organ Culture ◽

Tight Junctions ◽

Sertoli Cell ◽

Germ Cells ◽

Sertoli Cells ◽

Barrier Function ◽

Cell Junction ◽

Intramembranous Particles ◽

Junctional Complexes ◽

Oriented Parallel

The Sertoli cell tight junctional complexes have been shown to be the most important structural counterpart of the physiological blood-testis barrier. In freeze etch replicas they consist of extensive rows of intramembranous particles which are not only oriented parallel to one another, but to the myoid layer as well. Thus the occluding complex has both an internal and an overall orientation. However, this overall orientation to the myoid layer does not seem to be necessary to its barrier function. The 20 day old rat has extensive parallel tight junctions which are not oriented with respect to the myoid layer, and yet they are inpenetrable by lanthanum. The mechanism(s) for the control of Sertoli cell junction development and orientation has not been established, although such factors as the presence or absence of germ cells, and/or hormones, especially FSH have been implicated.

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