Differential expression of acidic cytokeratins 18 and 19 during sexual differentiation of the rat gonad

Development ◽  
1992 ◽  
Vol 115 (2) ◽  
pp. 503-517
Author(s):  
V. Fridmacher ◽  
O. Locquet ◽  
S. Magre
Keyword(s):  
Monoclonal Antibodies ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Sexual Differentiation ◽  
Somatic Cells ◽  
Positive Staining ◽  
Female Rat ◽  
Male And Female ◽  
Future Studies ◽  
Testicular Differentiation

The expression of cytokeratins (CKs) 8, 18 and 19 was analyzed in male and female rat gonads from the undifferentiated stage (12.5 days of gestation) until two weeks after birth by indirect immunofluorescence, using specific monoclonal antibodies anti-CK 8 (LE41), anti-CK 19 (LP2K) and anti-CK 18 (LE65 and RGE53). In the undifferentiated blastema, the somatic cells were stained for CK 8 and CK 19, whereas no detectable immunoreactivity for CK 18 was obtained. The same staining CK pattern was observed in ovaries, in the somatic cells of ovigerous cords and in primary follicles. The staining was progressively decreasing in growing follicles after one week after birth. At the onset of testicular differentiation, when the first Sertoli cells differentiate in the gonad of 13.5-day old male fetuses, positive staining for CK 18 became evident, in addition to CK 8 and CK 19 expression. In the following days, CK 8, CK 18 and CK 19 were detected in Sertoli cells in the differentiating seminiferous cords, but progressively the reactivity for CK 19 decreased and was no longer observed after 18.5-19.5 days of gestation. In all cases, CKs were found to be coexpressed with vimentin, and germ cells were negative for both vimentin and CKs. The results reported here show first, that CKs are expressed before sexual differentiation in gonadal blastema in which no epithelial organization is observed, and second, that there is a CK 18/CK 19 shift in expression during morphogenesis of the testis which is not observed in the differentiating ovary. Future studies will have to determine whether these differences in CK expression are due to epitope-masking phenomena or to the regulation of CK synthesis.

scholarly journals Abnormal Meiosis Initiation in Germ Cell Caused by Aberrant Differentiation of Gonad Somatic Cell

2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Min Chen ◽  
Min Chen ◽  
Suren Chen ◽  
Jingjing Zhou ◽  
Fangfang Dong ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Germ Cell ◽  
Somatic Cell ◽  
Germ Cells ◽  
Somatic Cells ◽  
Ectopic Expression ◽  
Genital Ridge ◽  
Male And Female ◽  
Germ Cell Differentiation ◽  
Exact Function

The interaction between germ cell and somatic cell plays important roles in germ cell development. However, the exact function of gonad somatic cell in germ cell differentiation is unclear. In the present study, the function of gonad somatic cell in germ cell meiosis was examined by using mouse models with aberrant somatic cell differentiation. In Wt1R394W/R394W mice, the genital ridge is absent due to the apoptosis of coelomic epithelial cells. Interestingly, in both male and female Wt1R394W/R394W germ cells, STRA8 was detected at E12.5 and the scattered SYCP3 foci were observed at E13.5 which was consistent with control females. In Wt1-/flox; Cre-ERTM mice, Wt1 was inactivated by the injection of tamoxifen at E9.5 and the differentiation of Sertoli and granulosa cells was completely blocked. We found that most germ cells were located outside of genital ridge after Wt1 inactivation. STRA8, SYCP3, and γH2AX proteins were detected in germ cells of both male and female Wt1-/flox; Cre-ERTM gonads, whereas no thread-like SYCP3 signal was observed. Our study demonstrates that aberrant development of gonad somatic cells leads to ectopic expression of meiosis-associated genes in germ cells, but meiosis was arrested before prophase I. These results suggest that the proper differentiation of gonad somatic cells is essential for germ cell meiosis.

scholarly journals IGSF11 is required for pericentric heterochromatin dissociation during meiotic diplotene

PLoS Genetics ◽  
2021 ◽  
Vol 17 (9) ◽  
pp. e1009778
Author(s):  
Bo Chen ◽  
Gengzhen Zhu ◽  
An Yan ◽  
Jing He ◽  
Yang Liu ◽  
...  
Keyword(s):  
Germ Cells ◽  
Sertoli Cells ◽  
Somatic Cells ◽  
Primary Spermatocyte ◽  
Pericentric Heterochromatin ◽  
Spermatogenic Cells ◽  
Testicular Cell ◽  
Reporter System ◽  
Fluorescent Reporter ◽  
Late Pachytene

Meiosis initiation and progression are regulated by both germ cells and gonadal somatic cells. However, little is known about what genes or proteins connecting somatic and germ cells are required for this regulation. Our results show that deficiency for adhesion molecule IGSF11, which is expressed in both Sertoli cells and germ cells, leads to male infertility in mice. Combining a new meiotic fluorescent reporter system with testicular cell transplantation, we demonstrated that IGSF11 is required in both somatic cells and spermatogenic cells for primary spermatocyte development. In the absence of IGSF11, spermatocytes proceed through pachytene, but the pericentric heterochromatin of nonhomologous chromosomes remains inappropriately clustered from late pachytene onward, resulting in undissolved interchromosomal interactions. Hi-C analysis reveals elevated levels of interchromosomal interactions occurring mostly at the chromosome ends. Collectively, our data elucidates that IGSF11 in somatic cells and germ cells is required for pericentric heterochromatin dissociation during diplotene in mouse primary spermatocytes.

scholarly journals Sertoli–germ cell junctions in the testis: a review of recent data

2010 ◽  
Vol 365 (1546) ◽  
pp. 1593-1605 ◽  
Author(s):  
Ilona A. Kopera ◽  
Barbara Bilinska ◽  
C. Yan Cheng ◽  
Dolores D. Mruk
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Seminiferous Epithelium ◽  
Cell Junctions ◽  
Molecular Architecture ◽  
Future Studies ◽  
Junction Type ◽  
Stage Viii ◽  
Immunological Barrier

Spermatogenesis is a process that involves an array of cellular and biochemical events, collectively culminating in the formation of haploid spermatids from diploid precursor cells known as spermatogonia. As germ cells differentiate from spermatogonia into elongated spermatids, they also progressively migrate across the entire length of the seminiferous epithelium until they reach the luminal edge in anticipation of spermiation at late stage VIII of spermatogenesis. At the same time, these germ cells must maintain stable attachment with Sertoli cells via testis-unique intermediate filament- (i.e. desmosome-like junctions) and actin- (i.e. ectoplasmic specializations, ESs) based cell junctions to prevent sloughing of immature germ cells from the seminiferous epithelium, which may result in infertility. In essence, both desmosome-like junctions and basal ESs are known to coexist between Sertoli cells at the level of the blood–testis barrier where they cofunction with the well-studied tight junction in maintaining the immunological barrier. However, the type of anchoring device that is present between Sertoli and germ cells depends on the developmental stage of the germ cell, i.e. desmosome-like junctions are present between Sertoli and germ cells up to, but not including, step 8 spermatids after which this junction type is replaced by the apical ES. While little is known about the biology of the desmosome-like junction in the testis, we have a relatively good understanding of the molecular architecture and the regulation of the ES. Here, we discuss recent findings relating to these two junction types in the testis, highlighting prospective areas that should be investigated in future studies.

scholarly journals Transcriptomic analysis of dead end knockout testis reveals germ cell and gonadal somatic factors in Atlantic salmon

2019 ◽  
Author(s):  
Lene Kleppe ◽  
Rolf Brudvik Edvardsen ◽  
Tomasz Furmanek ◽  
Eva Andersson ◽  
Kai Ove Skaftnesmo ◽  
...  
Keyword(s):  
Atlantic Salmon ◽  
Germ Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Somatic Cells ◽  
The Body ◽  
Genetic Introgression ◽  
Transcriptome Data ◽  
Dead End ◽  
Testis Tissue

Abstract Background Sustainability challenges are currently hampering an increase in salmon production. Using sterile salmon can solve problems with precocious puberty and genetic introgression from farmed escapees to wild populations. Recently sterile salmon was produced by knocking out the germ cell-specific dead end (dnd). Several approaches may be applied to inhibit Dnd function, including gene knockout, knockdown or immunization. Since it is challenging to develop a successful treatment against a gene product already existing in the body, alternative targets are being explored. Germ cells are surrounded by, and dependent on, gonadal somatic cells. Targeting genes essential for the survival of gonadal somatic cells may be good alternative targets for sterility treatments. Our aim was to identify and characterize novel germ cell and gonadal somatic factors in Atlantic salmon. Results We have for the first time analysed RNA-sequencing data from germ cell-free (GCF)/dnd knockout and wild type (WT) salmon testis and searched for genes preferentially expressed in either germ cells or gonadal somatic cells. To exclude genes with extra-gonadal expression, our dataset was merged with available multi-tissue transcriptome data. We identified 389 gonad specific genes, of which 194 were preferentially expressed within germ cells, and 11 were confined to gonadal somatic cells. Interestingly, 5 of the 11 gonadal somatic transcripts represented genes encoding secreted TGF-β factors; gsdf, inha, nodal and two bmp6-like genes, all representative vaccine targets. Of these, gsdf and inha had the highest transcript levels. Expression of gsdf and inha was further confirmed to be gonad specific, and their spatial expression was restricted to granulosa and Sertoli cells of the ovary and testis, respectively. Finally, we show that inha expression increases with puberty in both ovary and testis tissue, while gsdf expression does not change or decreases during puberty in ovary and testis tissue, respectively. Conclusions This study contributes with transcriptome data on salmon testis tissue with and without germ cells. We provide a list of novel and known germ cell- and gonad somatic specific transcripts, and show that the expression of two highly active gonadal somatic secreted TGF-β factors, gsdf and inha, are located within granulosa and Sertoli cells.

Regulation of meiosis in the foetal mouse gonad

Development ◽  
1982 ◽  
Vol 68 (1) ◽  
pp. 59-67
Author(s):  
Clive W. Evans ◽  
Diana I. Robb ◽  
Fiona Tuckett ◽  
Susan Challoner
Keyword(s):  
In Vitro Culture ◽  
Cross Section ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Inhibitory Effects ◽  
Culture Methods ◽  
Male And Female ◽  
Mixed Age ◽  
Gonadal Structure

In vitro culture of male and female gonads was found to have significant effects on gonadal structure and development. Culture resulted in a reduction of testicular cord diameter and a reduction in the number of Sertoli cells lining each cord in cross section. In the female, culture increased the percentage of pyknotic oocytes and fewer germ cells per unit of ovary volume reached diplotene. Mixed sex co-culture using different culture methods showed that day 14 p.c. testes inhibited meiosis in day 14 p.c. ovaries when the cultures were continued until the equivalent of day 21 p.c. Day 15 p.c. and mixed age co-cultures of mixed sex provided more equivocal data since meiosis was inhibited in some preparations but not in others. The possibility is suggested that prophase I may proceed irrevocably to diplotene after about day 15 p.c. and thus the inhibitory effects of foetal testes may be a function of female gonadal age. No evidence was found to support the hypothesis that mixed sex co-culture may stimulate meiosis precociously in foetal testes.

scholarly journals Spermatogenesis in sheep supplemented with detoxified castor bean (Ricinus communis L.) as a replacement for soybean meal

2020 ◽  
Vol 21 ◽  
Author(s):  
Laiara Fernandes ROCHA ◽  
Márcio de Oliveira RIBEIRO ◽  
Ana Lúcia Almeida SANTANA ◽  
Ronival Dias Lima de JESUS ◽  
Rosiléia Silva SOUZA ◽  
...  
Keyword(s):  
Germ Cells ◽  
Soybean Meal ◽  
Sertoli Cells ◽  
Castor Bean ◽  
Ricinus Communis ◽  
Panicum Maximum ◽  
Future Studies ◽  
Ricinus Communis L ◽  
The Mean ◽  
Level Of Significance

ABSTRACT The objective was to evaluate the effect of replacing soybean meal with the detoxified castor bean cake on testicular morphometry and spermatogenesis of sheep. Were used 24 uncastrated, 9-month old sheep weighing 29±0.8 kg they were randomly distributed among three treatments: T1 = 0%, T2 = 50%, and T3 = 100% substitution of soybean meal with detoxified castor bean cake. The animals were fed with Aruana grass pastage (Panicum maximum ‘Aruana’) and a ration for 90 days. After slaughtering, the testicles were collected and histological slides were prepared with tissue fragments. The data were evaluated for normality using the Shapiro-Wilk test, and analysis of variance was carried out at 5% level of significance. Substitution of soybean meal with detoxified castor bean cake had no effect on any of the assessed variables at the tested levels (P >0.05). The mean yield of spermatogenesis was 72.91 rounded spermatids per spermatogonium; the mean of total number of germ cells held by a Sertoli cell was 12.09; the mean of the testicular spermatic reserve was 31.82×109 and that per testicular gram was 238.28×106; the mean of daily spermatic production was 3.03×109 and that per testicular gram was 22.69×106; and the total number of Sertoli cells was 4.15×109 and that per testicular gram was 34.51×106. The results show that it is possible to replace 100% of the soybean meal with detoxified castor bean cake in sheep diet without any effects on spermatogenesis; however, it is important to perform seminal evaluations in future studies.

scholarly journals Transcriptomic analysis of dead end knockout testis reveals germ cell and gonadal somatic factors in Atlantic salmon

2020 ◽  
Author(s):  
Lene Kleppe ◽  
Rolf Brudvik Edvardsen ◽  
Tomasz Furmanek ◽  
Eva Andersson ◽  
Kai Ove Skaftnesmo ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Gene Knockout ◽  
Somatic Cells ◽  
The Body ◽  
Genetic Introgression ◽  
Transcriptome Data ◽  
Dead End ◽  
Testis Tissue

Abstract Background Sustainability challenges are currently hampering an increase in salmon production. Using sterile salmon can solve problems with precocious puberty and genetic introgression from farmed escapees to wild populations. Recently sterile salmon was produced by knocking out the germ cell-specific dead end (dnd). Several approaches may be applied to inhibit Dnd function, including gene knockout, knockdown or immunization. Since it is challenging to develop a successful treatment against a gene product already existing in the body, alternative targets are being explored. Germ cells are surrounded by, and dependent on, gonadal somatic cells. Targeting genes essential for the survival of gonadal somatic cells may be good alternative targets for sterility treatments. Our aim was to identify and characterize novel germ cell and gonadal somatic factors in Atlantic salmon.Results We have for the first time analysed RNA-sequencing data from germ cell-free (GCF)/dnd knockout and wild type (WT) salmon testis and searched for genes preferentially expressed in either germ cells or gonadal somatic cells. To exclude genes with extra-gonadal expression, our dataset was merged with available multi-tissue transcriptome data. We identified 389 gonad specific genes, of which 194 were preferentially expressed within germ cells, and 11 were confined to gonadal somatic cells. Interestingly, 5 of the 11 gonadal somatic transcripts represented genes encoding secreted TGF-β factors; gsdf, inha, nodal and two bmp6-like genes, all representative vaccine targets. Of these, gsdf and inha had the highest transcript levels. Expression of gsdf and inha was further confirmed to be gonad specific, and their spatial expression was restricted to granulosa and Sertoli cells of the ovary and testis, respectively. Finally, we show that inha expression increases with puberty in both ovary and testis tissue, while gsdf expression does not change or decreases during puberty in ovary and testis tissue, respectively.Conclusions This study contributes with transcriptome data on salmon testis tissue with and without germ cells. We provide a list of novel and known germ cell- and gonad somatic specific transcripts, and show that the expression of two highly active gonadal somatic secreted TGF-β factors, gsdf and inha, are located within granulosa and Sertoli cells.

scholarly journals Transgenic and knockout analyses of Masculinizer and doublesex illuminated the unique functions of doublesex in germ cell sexual development of the silkworm, Bombyxmori

2020 ◽  
Author(s):  
Tomohisa Yuzawa ◽  
Misato Matsuoka ◽  
Megumi Sumitani ◽  
Fugaku Aoki ◽  
Hideki Sezutsu ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sexual Development ◽  
Sexual Differentiation ◽  
Somatic Cells ◽  
Insect Species ◽  
Specific Factor ◽  
Homozygous Mutation ◽  
Rna Immunoprecipitation ◽  
Male Specific

Abstract Background Masculinizer (Masc) plays a pivotal role in male sex determination in the silkworm, Bombyx mori. Masc is required for male-specific splicing of B. mori doublesex (Bmdsx) transcripts. The male isoform of Bmdsx (BmdsxM) induces male differentiation in somatic cells, while females express the female isoform of Bmdsx (BmdsxF), which promotes female differentiation in somatic cells. However, the importance of Bmdsx in sexual differentiation in germ cells remains unclear. In Drosophila melanogaster, mechanisms regulating sexual differentiation differ between germ cells and somatic cells. dsx is not required within female or male germ cells for sexual development. However, it remains unclear whether this is also the case in other insect species. Results In this study, we performed genetic analyses using a transgenic line that expressed Masc, as well as various Bmdsx knockout lines. Masc-expressing females express both BmdsxF and BmdsxM and have degenerated ovaries combined with testis-like tissues, which produce sperm. We found that Masc-expressing females with a homozygous mutation in BmdsxM showed normal development in ovaries. The formation of testis-like tissues was abolished in these females. In comparison, Masc-expressing females carrying a homozygous mutation in BmdsxF exhibited almost complete male-specific development in gonads and germ cells. These results suggest that BmdsxM can induce male development in germ cells and internal genital organs, while BmdsxF inhibits BmdsxM activity and represses male differentiation. To investigate whether MASC directly controls male-specific splicing of Bmdsx and identify RNAs that form complexes with MASC in testes, we performed RNA immunoprecipitation (RIP) using an anti-MASC antibody. We found that MASC formed a complex with Bmdsx-AS1 lncRNA, which is a testis-specific factor involved in the male-specific splicing of Bmdsx pre-mRNA. Conclusions Combined, our findings suggest that Masc induces male differentiation in gonads and germ cells by interacting with Bmdsx-AS1 lncRNA and enhancing the production of BmdsxM. Unlike Drosophila dsx, BmdsxM was able to induce spermatogenesis in genetically female (ZW) germ cells. To our knowledge, this is the first report indicating the role of dsx in germ cell sexual development differs among insect species.

scholarly journals Immunocytochemical observation of the 90 KD heat shock protein (HSP90): high expression in primordial and pre-meiotic germ cells of male and female rat gonads.

1995 ◽  
Vol 43 (1) ◽  
pp. 67-76 ◽  
Author(s):  
S Ohsako ◽  
D Bunick ◽  
Y Hayashi
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Gonad Development ◽  
Testicular Development ◽  
Female Rat ◽  
Male And Female ◽  
Primordial Follicles ◽  
Pachytene Spermatocytes

We used immunocytochemistry to detect the 90 KD major heat shock protein (HSP90), a potential regulator of gene expression, during male and female rat gonad development. In the Day 13.0 post-coital (dpc) fetal gonad, strong immunoreactivity to anti-HSP90 antibody was shown in the cytoplasm of primordial germ cells (PGCs). Other somatic cells in the gonad showed only faint reactivity. During testicular development, strong immunostaining was observed in the cytoplasm of embryonic germ cells and in spermatogonia and spermatocytes of the pre-pubertal testis. In adult testis reactivity of spermatogonia and pachytene spermatocytes was strong but reactivity of post-meiotic spermatogenic cells, i.e., secondary spermatocytes and spermatids, was extremely reduced. During ovarian development, immunostaining was also observed in the oogonia and the oocytes of pre-pubertal ovary. However, the staining of oocytes was reduced with the development of primordial follicles during the first week after birth. This study revealed that HSP90 is highly expressed in PGCs and continues to be expressed in both male and female pre-meiotic germ cells. The HSP90 accumulation may be essential for both male and female mammalian pre-meiotic germ cells.

scholarly journals Tight junctions in the testis: new perspectives

2010 ◽  
Vol 365 (1546) ◽  
pp. 1621-1635 ◽  
Author(s):  
Dolores D. Mruk ◽  
C. Y. Cheng
Keyword(s):  
Epithelial Cells ◽  
Cancer Cells ◽  
Tight Junctions ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Systemic Circulation ◽  
Signalling Pathways ◽  
Future Studies ◽  
Cellular Event ◽  
Stage Viii

In the testis, tight junctions (TJs) are found between adjacent Sertoli cells at the level of the blood–testis barrier (BTB) where they coexist with basal ectoplasmic specializations and desmosome-gap junctions. The BTB physically divides the seminiferous epithelium into two distinct compartments: a basal compartment where spermatogonia and early spermatocytes are found, and an adluminal compartment where more developed germ cells are sequestered from the systemic circulation. In order for germ cells (i.e. preleptotene spermatocytes) to enter the adluminal compartment, they must cross the BTB, a cellular event requiring the participation of several molecules and signalling pathways. Still, it is not completely understood how preleptotene spermatocytes traverse the BTB at stage VIII of the seminiferous epithelial cycle. In this review, we discuss largely how TJ proteins are exploited by viruses and cancer cells to cross endothelial and epithelial cells. We also discuss how this information may apply to future studies investigating the movement of preleptotene spermatocytes across the BTB.

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