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At least 19 recordsLinked to original sources
PubMed1979

Characterization of Sertoli cell-germ cell junctional specializations in dissociated testicular cells.

To further characterize Sertoli cell-germ cell junctional specializations seminiferous tubules from sexually mature Sprague-Dawley rats were dissociated by enzymatic and mechanical methods. Ultrastructural analysis of cell suspensions prepared by incubation in collagenase alone or by mechanical methods revealed that spermatids remained attached to Sertoli cells or Sertoli cell fragments. Such cellular associations were found only between Sertoli cell fragments and spematids in which the developing acrosome had made contact with the plasma membrane (step 8 and subsequent steps of spermiogenesis). Furthermore, the fragments were confined to that region of the plasma membrane over the acrosome. The Sertoli cell half of this adhesive site displayed the typical elements of Sertoli cell junctions, filamentous bundles and associated cisterna of endoplasmic reticulum, in apposition to the spermatids. The spermatids demonstrated no surface specializations at the attachment sites. In contrast, in cell suspensions prepared with trypsin, spermatids were free of attachments to Sertoli cells or their fragments. These results demonstrate that: (1) the junctions act to bind cells together, (2) adhesive type contact is established between Sertoli cells and spermatids at step 8 and subsequent steps of spermiogenesis, (3) contact is restricted to the spermatid plasma membrane over the acrosome, and (4) spermatids can be freed from the junctional specializations by treatment with trypsin.

Animals↗
PubMed1992

[Studies on the aggregation of rat Sertoli cells, germ cells and peritubular cells in vitro].

Dissociated testicular cell suspensions from 5, 10, 15, 20 and 25 day postnatal rats were cultured either on solid agar as a substratum or in a flask under slow rotation. Spherical or cord-like aggregations were formed from cells under both conditions. Sections from the aggregations showed that they were comprised mainly of Sertoli cells, with peritubular cells located around the aggregations. A few germ cells were present within those aggregations from cultures of rat testicular cells younger than ten days, while no germ cell could be found within aggregations from rats older than fifteen days. Based on these observations, we conclude that testicular cells in post-natal rats still maintain a limited capacity for cell adhesion and recognition and are thus able to form cellular aggregations.

Animals↗
PubMed1992

Quantitation of Sertoli cell-germ cell desmosome gap junctions in relation to meiotic divisions in the male rat.

Desmosome-gap (D-G) junctions were quantified in relation to germ cell meiosis in the male, specifically to test the hypothesis that the loss of these junctions is related to successful passage of cells through diplotene phase of Meiosis I and the two cytokineses that follow. Such a hypothesis has been proposed as the cause for the resumption of meiosis that occurs prior to ovulation in the female. D-G junctions were quantified in pachytene spermatocytes (stage XII), diplotene spermatocytes (stage XII), secondary spermatocytes (stage XIV) and step 1 spermatids (stage I). These were referred to as the cells of interest as compared with spermatocytes (zygotene spermatocytes, zygotene spermatocytes, pachytene spermatocytes, pachytene spermatocytes) in the same stages, respectively, that served as controls termed control cells. Since gap junctions are not easily recognized in the average sectioned profile of a desmosome-gap junction, only the desmosomal component was quantified. The data were expressed as both numbers and length of junctions per tubule, per cell profile and per unit lineal membrane length to overcome errors inherent in the methodologies utilized. There was no indication that numbers of junctions changed specifically in the cells of interest after passage through diplotene suggesting that these junctions do not have a comparable role in meiotic continuance in the male as proposed for the female. Interestingly, the control cells always showed greater numbers and length of junctions than the cells of interest suggesting that junction may relate more to the period of initiation of meiosis than to its continuance.

Animals↗
PubMed1978

Occurrence of nuage in fetal human germ cells.

Germ cells of human fetuses aged 7-13 weeks were studied. Dense fibrous material called nuage was found in the cytoplasm of human fetal germ cells. The nuage increased in frequency in the germ cells during the interval studied. In oogonia the nuage occurred much more frequently than in spermatogonia and the frequency of nuage in oogonia increased earlier than in spermatogonia. The number of nuage was continuously at a higher level in oogonia than in spermatogonia. It is suggested that these findings support the theory that the nuage may act as a germ cell determinant.

Female↗
PubMed1979

Interrelationships between Sertoli cells and germ cells in the Syrian hamster.

The interrelationships of the Sertoli cells and germ cells in the Syrian hamster were examined using the electron microscope. Demosome-like junctions were observed attaching Sertoli cells to spermatogonia and spermatocytes. In the region of the junctions dense plaques lay on the cytoplasmic surfaces of the plasmalemma of the opposing cells. Sertoli cell cytoplasmic filaments converged in the area of the junctions and inserted into the subsurface densities. Filaments were not observed associated with the subsurface densities of the germ cells. In the region of the junctions a 15...20 nm gap, filled with an attenuate amorphous substance, separated the plasmalemmata. Another attachment device termed "junctional specialization" occurred between Sertoli cells, and preleptotene spermatocytes and all successive developmental steps in the germ cell line in the hamster. The junctional specializations consisted of a mantel of Sertoli cell cytoplasmic filament lying subjacent to the Sertoli cell plasmalemma and an opposed cisterna of the endoplasmic reticulum. In stages VII-VIII preleptotene supermatocytes were observed in transit from the basal compartment to the adluminal compartment. While Sertoli-Sertoli junctions adluminal to the spermatocytes remained intact, typical Sertoli-Sertoli junctions formed between opposed Sertoli cell processes basal to the spermatocytes. It is proposed that, during the passage of spermatocytes in to the adluminal compartment, junctional specializations associated with preleptotene spermatocytes in the basal compartment migrate basal to the spermatocytes and contribute to formation of Sertoli-Sertoli junctions. Treatment of seminiferous tubules with hypertonic media was used to demonstrate that the junctional specializations function in cell-to-cell adhesion. Data indicated that these junctions function to retain the developing spermatids within the seminiferous epithelijm until the time of spermiation. At spermination the junctional specializations disappear and the spermatids drift off into the tubule lumen.

Animals↗
PubMed2026

Overlapping genetic etiology of pediatric and adult germ cell tumors.

BACKGROUND: Germ cell tumors are heterogeneous neoplasms arising from primordial germ cells. Although genome-wide association studies have identified numerous susceptibility loci for adult testicular germ cell tumors, the heritable basis of pediatric testicular germ cell tumors and germ cell tumors that arise outside the testes remain poorly understood. METHODS: We conducted a multi-ancestry genome-wide association study of pediatric germ cell tumors, including 1927 cases from the Germ Cell Tumor Epidemiology Study and 10 601 controls. Cases were diagnosed with testicular (n = 678), ovarian (n = 441), intracranial (n = 435), and extragonadal (n = 373) germ cell tumor between the ages of 0 and 19 years. RESULTS: We identified 4 loci reaching genome-wide significance, including variants near BAK1 (chr 6: rs3831846), SPRY4 (chr 5: rs12515244), DMRT1 (chromosome [chr] 9: rs10815910), and DEPTOR (chr 8: rs13277786). Additional genome-wide statistically significant associations were identified in subgroup analyses, including 6 loci for intracranial germ cell tumors (rs2758612 [PMF1/BGLAP], rs9854760 [PLCL2], rs6851498 [KIT], rs11816992 on chromosome 10, rs3830273 [TFAM], and rs13054014 [LZTR1]), 1 locus for testicular germ cell tumor (rs1907702 [KITLG]), and 1 locus for males (rs4610628 [MAD1L1]). After Bonferroni correction, 18 of 78 previously reported testicular germ cell tumor loci were significantly associated with germ cell tumor overall or in at least 1 subgroup with a particularly strong correlation between testicular germ cell tumor and intracranial germ cell tumor effect estimates (rho = 0.63, P = 5.5 × 10-10). Expression quantitative trait locus (QTL) analyses identified candidate genes in the regions identified on chromosome 6 (BAK1, LINC003366, and ITPR3) and chromosome 8 (DEPTOR and RP11-760H22.2). CONCLUSIONS: Our data support a role for germline genetic variation in the development of germ cell tumors in locations outside the testes and highlight shared genetic architecture across age group and tumor location.

Humans↗
PubMed1992

Chemotherapy of male and female germ cell tumors.

Germ cell tumors of testicular origin are virtually always curable with modern therapy that appropriately integrates chemotherapy and surgery. Clinical trials of chemotherapy in patients with disseminated disease now separate those with limited metastatic disease and an excellent prognosis from those with bulky tumor and a less favorable prognosis. In the former group, it has been shown that three courses of BEP (bleomycin/etoposide/cisplatin) are therapeutically comparable to four courses. The next generation study compared BEP with a similar regimen with the bleomycin omitted. Early results of this study suggest that deletion of bleomycin worsens outcome. Other studies have addressed the same question but have used four courses of this agent. Ovarian germ cell tumors have been less well studied but are largely curable with surgery and chemotherapy. The specific regimens used are similar to their testis tumor counterparts. All patients with completely resected tumor should receive adjuvant chemotherapy; virtually all will remain cancer free. Of course, patients with advanced disease should also receive chemotherapy. Results are generally good in these patients, but probably less so than in testis tumor patients with similar tumor volume.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1975

Chemical induction of presumed dominant-lethal mutations in postcopulation germ cells of mice. I. Relative sensitivity between pre- and postcopulation germ cells to isopropyl methanesulfonate.

Females from (C3H X 101)F1 and a mixed stock were injected intraperitoneally with either 25 or 50 mg/kg isopropyl methanesulfonate (IMS) or Hanks' solution 4-5 h after the midpoint of the dark period during which mating occurred. It was determined that at the time of treatment the great majority of oocytes were undergoing second meiotic division. For comparison, the same doses of IMS were given to females treated within 3-5 days prior to mating (predominantly dictyate oocytes) or to males treated within 4-5 days prior to mating (sperm in vas and epididymis). The frequencies of presumed dominant lethals induced by 50 mg/kg IMS in sperm treated in vas and epididymis, dictyate oocytes, and germ cells in mated females are 22%, 19%, and 79%, respectively, for (C3H X 101)F1 and 26%, 30%, and 76% for the other stock. Clearly, in both stocks, effects in mated females, when both female and male germ cells were treated, are relatively much higher than the added effects on dyctyate oocytes and spermatozoa. This is also true for the 25 mg/kg dose.

Animals↗
PubMed1992

Sterol carrier protein 2 (non-specific lipid transfer protein) is localized in membranous fractions of Leydig cells and Sertoli cells but not in germ cells.

The cellular and subcellular distribution of sterol carrier protein 2 (SCP2; nsL-TP) was reinvestigated in rat testicular cells by Western blotting and immunocytochemistry, using the affinity purified antibody against rat liver SCP2. Western blot analysis revealed high levels of the protein in the somatic cells of the testis, e.g., Leydig and Sertoli cells whereas it could not be detected in germ cells. This cellular localization of SCP2 was confirmed by Northern blotting. Immunocytochemical techniques revealed that in Leydig cells, immunoreactive proteins were concentrated in peroxisomes. Although SCP2 was also detected in Sertoli cells, a specific subcellular localization could not be shown. SCP2 was absent from germ cells. Analysis of subcellular fractions of Leydig cells showed that SCP2 is membrane bound without detectable amounts in the cytosolic fraction. These results are at variance with data published previously which suggested that in Leydig cells a substantial amount of SCP2 was present in the cytosol and that the distribution between membranes and cytosol was regulated by luteinizing hormone. The present data raise the question in what way SCP2 is involved in cholesterol transport between membranes in steroidogenic cells but also in non-steroidogenic cells.

Animals↗
PubMed1992

Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture.

Steel factor (SF) and LIF (leukemia inhibitory factor) synergistically promote the proliferation and survival of mouse primordial germ cells (PGCs), but only for a limited time period in culture. We show here that addition of bFGF to cultures in the presence of membrane-associated SF and LIF enhances the growth of PGCs and allows their continued proliferation beyond the time when they normally stop dividing in vivo. They form colonies of densely packed, alkaline phosphatase-positive, SSEA-1-positive cells resembling undifferentiated embryonic stem (ES) cells in morphology. These cultures can be maintained on feeder layers for at least 20 passages, and under appropriate conditions give rise to embryoid bodies and to multiple differentiated cell phenotypes in monolayer culture and in tumors in nude mice. PGC-derived ES cells can also contribute to chimeras when injected into host blastocysts. The long-term culture of PGCs and their reprogramming to pluripotential ES cells has important implications for germ cell biology and the induction of teratocarcinomas.

Alkaline Phosphatase↗
PubMed1992

Mediastinal germ cell tumors.

Mediastinal germ cell tumors are among the most common tumors in the anterior mediastinum. This tumor should be considered strongly in the differential diagnosis of anterior mediastinal masses. Benign teratomas are common in this site, and complete surgical resection results in cure of nearly all patients. Malignant mediastinal seminomas are highly curable and either radiotherapy (mediastinal disease) or cisplatin-based chemotherapy (metastatic disease) will result in long-term survival in 80% or more of patients. Mediastinal nonseminomatous germ cell malignancies represent a clinically and biologically distinct subset of mediastinal germ cell neoplasms. Relative to their testicular counterparts, these tumors carry a poor prognosis, but, with newer refinements in cisplatin-based chemotherapy, approximately half of these patients will survive their illness. The history of success with chemotherapy should prompt thorough pathologic and serologic evaluation of all patients with mediastinal malignancies in hopes of defining a curable process. In particular, poorly differentiated carcinomas at this site should be treated as germ cell tumors, and so long-term survival will be attainable. Mediastinal nonseminomatous germ cell tumors are associated with the development of nongerm cell malignancies such as embryonal rhabdomyosarcomas and hematologic malignancies such as acute megakaryocytic leukemia and malignant histiocytosis. The development of these associated malignancies is not related to therapy and represents a true biological link between these malignancies. It is likely that the development of these malignancies is an expression of the multipotential nature of primitive germ cells. Careful clinical and biologic investigations of these rare transformations may lead to greater understanding of the regulation of events in the malignant process.

Humans↗
PubMed1992

A method to obtain avian germ-line chimaeras using isolated primordial germ cells.

Primordial germ cells (PGCs), collected from the blood of 2-day-old chick embryos, were concentrated by Ficoll density centrifugation. The blood contained 0.048% PGCs and the concentrated fraction contained 3.9% PGCs in blood cells. The PGCs were picked up with a fine glass pipette, and one hundred were then injected into the terminal sinuses of 2-day-old Japanese quail embryos (24 somites); bubbles were then inserted to prevent haemorrhage. The embryos were further incubated at 38 degrees C for 24 h, and then fixed. Serial sections were stained with the periodic acid-Schiff reagent (PAS) to demonstrate chicken PGCs and with Feulgen stain to identify quail cells. On the basis of the differences in staining properties, 63.6 +/- 5.3 chick PGCs were detected in the quail embryo in the area where the gonads develop. Furthermore, 39.3 +/- 4.5 chick PGCs were incorporated into the quail germinal epithelium within 24 h of the injection. A similar percentage of the host (quail) PGCs had also migrated to the germinal epithelium at the same stage of development. This technique for obtaining germ-line chimaeras will facilitate research on avian germ-line differentiation.

Animals↗
PubMed1992

Analysis of chromosome 12 aneuploidy in interphase cells from human male germ cell tumors by fluorescence in situ hybridization.

The i(12p) marker chromosome has been found to be a highly nonrandom chromosome abnormality associated with germ cell tumors (GCTs). We have previously shown that a chromosome 12 centromere specific alpha-satellite DNA probe detects the i(12p) by virtue of differences in the size of the signal originating from the i(12p) and normal chromosome 12 centromeres after fluorescence in situ hybridization (FISH) in metaphase and interphase cells of cultured GCT cell lines. We have now extended this analysis to 72 fresh GCT tumor biopsy specimens. Banded cytogenetic analysis was attempted on each of these tumors, 45 of which were found to be clonally abnormal. Data on i(12p) and chromosome 12 copy number obtained by FISH agreed well with those obtained by cytogenetic analysis. In addition, the FISH method made possible the detection and determination of i(12p) and the chromosome 12 copy number in cases in which conventional cytogenetic analysis was unsuccessful. We found the incidence of i(12p) in seminomas to be low (7%) compared to that in nonseminomas (75%) when tumor biopsy specimens were studied by FISH. Our results show that the FISH technique can be used reliably for detection of the diagnostically and prognostically useful i(12p) marker in GCT tumor biopsy specimens.

Aneuploidy↗
PubMed1977

Desmosome-like junctions between Sertoli and germ cells in the rat testis.

Desosome-like junctions between Sertoli cells and germ cells (spermatogonia, spermatocytes and non-elongate spermatids) were observed in the adult rat testis. At all levels, certain features were characteristic of this relationship, the most prominent being a densification of the subsurface aspect of the plasma membranes of each cell. The Sertoli cell counterpart demonstrated an abundance of cytoplasmic fibrils which converged on the density, but no such feature was evident in the germ cell counterpart. The width of the intercellular space was variable, ranging in some regions from approximately 3-5 nm, to other regions of approximately 14-18 nm, the latter being representative of most areas of contact. A poorly represented, and often discontinuous, intermediate dense line was observed in the intercellular space. Hypertonic fixative solutions containg dextrose were utilized to provoke tissue shrinkage and exaggeration of the intercellular space within the basal compartment of the testis. Cell separation was evident over most regions of the cells, except where desmosome-like contacts were present. Forces transmitted from one cell to another at desmosome-like contacts resulted in tearing of cellular fragments, while junctional regions of both cells remained intact. These observations indicate that desmosome-like junctions are strong adhesive sites between germ cells and Sertoli cells.

Animals↗
PubMed1992

Immunohistochemical and biochemical characterisation of the expression of a human embryonal carcinoma cell proteoglycan antigen in human germ cell tumours and other tissues.

In the embryonal carcinoma (EC) cell line GCT 27, monoclonal antibody GCTM-2 recognises an epitope on a 200 kD pericellular matrix keratan sulphate proteoglycan. Immunohistochemical analyses demonstrated staining of tissue sections from 21 out of 22 human non-seminomatous germ cell tumours, and from 22 out of 28 sections of seminomas. In normal human fetal tissues gut epithelium and muscle stained strongly, and certain other epithelia stained moderately. In adult tissues, the distribution of the epitope was similar, but staining intensity was weaker. Neoplastic tissues showed reactivity with embryonal rhabdomyosarcoma and colorectal carcinoma, but no other non-germ cell tumours. Immunofluorescence microscopy showed that GCTM-2 also stained cell lines from human colorectal carcinoma, embryonal rhabdomyosarcoma and choriocarcinoma. In contrast to EC cells the epitope in these other cell types required permeabilization of the cells to be visualised, and the protein bands in immunoblots lacked extensive modification with keratan sulphate and were smaller. Thus, GCTM-2 reacts with an epitope which has a previously unrecognised tissue distribution; its expression as a pericellular matrix proteoglycan is predominantly a characteristic of human EC cells.

Antibodies, Monoclonal↗