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A McLaren 《Canadian Metallurgical Quarterly》1998,42(7):855-860
The germ cell lineage in mice is established about a week after fertilization, in a group of cells that have left the epiblast and moved to an extraembryonic site. They migrate back into the embryo, along the hind gut and into the gonads. Germ cells in male and female embryos then pursue different pathways: in the testis the germ cells cease proliferating and enter mitotic arrest, while germ cells in the ovary, like those in male embryos that remain outside the gonads, enter meiotic prophase. Studies on explanted germ cells suggest that all germ cells may enter meiosis at a certain stage of their development, unless prevented from doing so by some inhibitory influence of the testis. Germ cells during the migratory stage can be cultured, but do not enter meiosis unless embedded in somatic tissue. Addition of certain growth factors and cytokines to the culture medium allows germ cells to proliferate indefinitely in vitro: Like embryonic stem cells, these immortalized EG (embryonic germ) cells will colonize all cell lineages if introduced into a blastocyst. After birth, germ cells undergo gametogenesis; oogenesis in the female, spermatogenesis in the male. Brinster and his colleagues have shown that spermatogonial stem cells injected into a germ-cell depleted testis will repopulate the seminiferous tubules and undergo spermatogenesis, giving rise to functional spermatozoa. Stem cells from frozen testicular tissue are still capable of giving rise to spermatogenesis in a host testis. Rat testicular tissue can undergo spermatogenesis in a mouse testis, to form morphologically normal rat spermatozoa, even though the Sertoli cells that support them are of endogenous mouse origin. These findings are of fundamental importance for our understanding of spermatogenesis and the interactions between germ cells and Sertoli cells; but they also have significant practical implications, in relation to both agricultural practice and clinical treatment of infertility. 相似文献
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Oocytes from many invertebrates initiate development within distinctive cysts of interconnected cells, which are formed through synchronous divisions of a progenitor cell. Recently, processes underlying cyst formation have been extensively characterized at the molecular level in Drosophila. Defects in this process cause sterility in female flies. Early female mouse germ cells are organized as cell clusters as well, but it is uncertain whether these groups are similar to the cysts of invertebrates. We find that mouse germ cells are connected by intercellular bridges in the ovaries of 11.5 to 17.5 days postcoitum embryos; microtubules and organelles have been observed within these bridges. Confocal microscopy shows that cells within mouse clusters divide synchronously and frequently correspond in number to powers of two. Thus, female mouse germ cell clusters exhibit key characteristics of invertebrate germline cysts indicating that the process of germline cyst formation is conserved in the mouse. 相似文献
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Quasi-homogeneous fractions of male mouse germ cells at definite stages of meiosis and spermiogenesis were obtained by using a separation method based on sedimentation velocity in an albumin gradient. In the various cell types, the total DNA-dependent DNA polymerase activity was determined, and the major enzymatic forms were characterized. The DNA polymerase species present in premeiotic, meiotic and post-meiotic cells were analyzed by glycerol gradient sedimentation. Two types of DNA polymerase were identified in fractions enriched in spermatogonia and preleptotene spermatocytes. One showed a sedimentation coefficient of about 7.5 S and was sensitive to N-ethylmaleimide (NEM); the other exhibited a sedimentation coefficient between 3 and 4 S and was resistant to NEM. On the basis of their sedimentation coefficients, their sensitivity to NEM and their template specificities, these 2 enzymes were identified respectively as alpha and beta DNA polymerases as reported in mammals. The gradient analysis performed on fractions enriched in meiotic and post-meiotic cells revealed the presence of DNA polymerase beta only. A quantitative analysis showed that the activity of the DNA polymerase beta reaches a maximum at middle-late pachytene stage and then drops gradually during spermiogenesis. Although any conclusion as to the biological role of this high level of DNA polymerase activity in pachytene spermatocytes is premature, it is tempting to suggest that this enzyme is involved in meiotic recombination. 相似文献
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C Sweeney M Murphy M Kubelka SE Ravnik CF Hawkins DJ Wolgemuth M Carrington 《Canadian Metallurgical Quarterly》1996,122(1):53-64
We have shown previously that a novel cell cycle-regulated histone H1 kinase activity, retinoblastoma kinase (RbK), associates with and phosphorylates the amino terminus of the Rb protein in G2-M. We have shown also that the amino terminus of p107, a Rb-related protein, does not associate with a similar kinase in vitro or in vivo. Here, we report that a RbK-like kinase associates with the amino terminus of p130, another Rb-related protein, only marginally. Moreover, the association of RbK with Rb in vitro is shown to require a discrete portion of the Rb amino terminus, amino acids 89-202. This region has been shown previously to be subject to inactivating mutations in retinoblastoma and to be required for Rb-mediated growth suppression in vitro. Taken together, these data indicate that the formation of Rb-RbK complexes may play an important role in Rb-mediated growth suppression. We have mapped two in vitro sites of Rb phosphorylation by RbK to sites that are phosphorylated in vivo and are targets of cyclin-dependent kinase phosphorylation in vitro. As such, at least some sites of RbK phosphorylation overlap with those of other proline-directed serine and threonine kinases. Consistent with this latter observation, we report that the trans-activation domain of c-myc is phosphorylated specifically by RbK in vitro at a site (serine 62) that is phosphorylated in vivo during G2-M, cell-cycle phases in which RbK activity is maximal. 相似文献
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OBJECTIVE: To determine the utility of CT-determined main pulmonary artery diameter (MPAD) for predicting pulmonary hypertension (PH) in patients with parenchymal lung disease. DESIGN: Retrospective review of right-heart hemodynamic data and chest CT scans in 45 patients. SETTING: Tertiary-referral teaching hospital and VA hospital. PATIENTS: Between October 1990 and December 1995, 36 patients referred for evaluation of parenchymal lung disease or possible pulmonary vascular disease were found to have PH, as defined by mean pulmonary artery pressure (mPAP) > or =20 mm Hg. Nine control patients (mPAP <20 mm Hg) were also identified (4 from hospital records search, 5 after evaluation for possible PH). RESULTS: CT-determined MPAD was 35+/-6 mm in patients with PH and 27+/-2 mm in control subjects. In our group of patients, MPAD > or =29 mm had a sensitivity of 87%, specificity of 89%, positive predictive value (PPV) of 0.97, and positive likelihood ratio (LR) of 7.91 for predicting PH; in the subgroup of patients with parenchymal lung disease (n=28, PH and control subjects), MPAD > or =29 mm had a sensitivity of 84%, specificity of 75%, PPV of 0.95, and positive LR of 3.36 for predicting PH. The most specific findings for the presence of PH were both MPAD > or =29 mm and segmental artery-to-bronchus ratio > 1:1 in three or four lobes (specificity, 100%). There was no linear correlation between the degree of PH and MPAD (r=0.124). CONCLUSIONS: CT-determined MPAD has excellent diagnostic value for detection of PH in patients with advanced lung disease. Therefore, standard chest CT scans can be used to screen for PH as a cause of exertional limitation in patients with parenchymal lung disease. Because CT is commonly used to evaluate parenchymal lung disease, this information is readily available. 相似文献
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BJ Gulyas 《Canadian Metallurgical Quarterly》1976,147(2):203-218
The fine structural changes were studied in the plasma membrane, cortical granules (CGs) and meiotic spindle of rabbit, hamster and mouse eggs in response to electrical stimulation. Eggs were collected 16 to 18 hours after HCG injection, and freed from the cumulus oophorus. They were stimulated in vitro by delivering a single monophasic square wave pulse of 150 V for 1 msec. Stimulated and unstimulated eggs were fixed for fine structural observations 1, 30 and 60 minutes after stimulation. Within one minute of stimulation the microvilli of rabbit eggs were long, branching and had bulbous ends. Umbonate protrusions were also present on their surface. By 30 minutes after stimulation the rabbit eggs lacked microvilli and the perivitelline space with detached vesicles. Degenerating changes were readily noticeable in the microvilli of hamster eggs by 60 minutes. Changes were not noted in the microvilli of stimulated mouse eggs. There were markedly fewer CGs in the hamster and mouse, but not rabbit, eggs by 30 minutes after stimulation, and nearly all of them disappeared in the mouse and hamster eggs by 60 minutes. The density of the CGs in the rabbit was not altered. The meiotic spindle of hamster eggs dissipated within one minute of stimulation, however, the microtubules reappeared by 60 minutes after stimulation. Rotation of the meiotic spindle occurred in the mouse and hamster eggs by 30 minutes after stimulation. In some of the mouse eggs the spindle migrated to the center and the chromosomes were in telophase. 相似文献
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E Archimbaud 《Canadian Metallurgical Quarterly》1997,39(5):264-268
We have evaluated whether low density lipoprotein (LDL) receptor activity of stimulated lymphocytes, as measured by an improved flow cytometric assay, may be used to diagnose familial hypercholesterolemia (FH). Cells were isolated from 75 children suspected from strict clinical criteria to be FH heterozygotes and from 29 normal children. DNA from the FH patients were also subjected to molecular genetic analysis of the LDL receptor gene in order to confirm the clinical diagnosis. A molecular genetic diagnosis of FH was obtained in 68 of the 75 patients; 67 of these had a low (below 70% of normal) receptor activity and 1 had a borderline (71%) activity. By contrast, 28 of the normal children showed a normal (above 80%) and 1 a borderline (78%) receptor activity. Of the 7 patients in whom no mutation in the LDL receptor gene was found, 4 showed a normal, 1 a borderline, and 2 showed a low activity. In summary, measurement of LDL receptor activity allowed us to separate between genetically diagnosed FH heterozygotes and healthy children. The combined use of LDL receptor activity measurements and molecular genetic analysis allows us both to diagnose and exclude FH in children suspected to suffer from this disease. 相似文献
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We have demonstrated the utility of ultrasound backscatter microscopy for targeted intraparenchymal injections into embryonic day (E) 13.5 mouse embryos. This system has been used to test the degree of commitment present in neural progenitors from the embryonic ventral telencephalon and mid-hindbrain region. Many E13.5 ventral telencephalic progenitors were observed to integrate and adopt local phenotypes following heterotopic transplantation into telencephalic or mid-hindbrain targets, whereas mid-hindbrain cells of the same stage were unable to integrate and change fate in the telencephalon. In contrast, many mid-hindbrain cells from an earlier developmental stage (E10.5) were capable of integrating and adopting a forebrain phenotype after grafting into the telencephalon, suggesting that mouse mid-hindbrain progenitors become restricted in their developmental potential between E10.5 and E13.5. 相似文献
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The incidence of aneuploidy in male germ cells was evaluated by analyzing extra marker chromosome(s) signal(s) in round and/or hook spermatids of transgenic mice. Two types of transgenic mice were used as models. The inserted foreign DNA (lambda-gt10LacZ shuttle vector and/or pSVc-myc plasmid) was located at the middle of the long arms of chromosome 2 (lambda DNA) and/or chromosome 8 (c-myc). The number of marker chromosomes present could easily be detected after fluorescence in situ hybridization (FISH) in testicular cells. The frequency of spontaneous aneuploidy of chromosome 2 was similar in round spermatids of lambda and lambda-myc mice. Differential involvement of chromosomes 2 and 8 was observed in both round and hook spermatids. The frequency of spontaneous aneuploidy in round spermatids was higher than that in hook spermatids. The frequency of aneuploidy of marker chromosomes was significantly higher in older mice (2 years old) than in younger ones. Diethylstilbestrol (DES)-induced aneuploidy was dose dependent, and was not influenced by the stage at which germ cells were treated with DES. These results demonstrate the usefulness of a transgenic mouse model for the study of aneuploidy in germ cells. 相似文献
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S Kawakami K Mitsunaga YY Kikuti A Ando H Inoko K Yamamura K Abe 《Canadian Metallurgical Quarterly》1998,9(11):874-880
Tctex3 showing restricted expression in male germ cells has been isolated during the process of chromosome walking in the mouse t-complex region. The total sequence of Tctex3 cDNA predicts a protein of 580 amino acids with two C4HC3 type PHD fingers. The region containing this conserved motif is shared among members of the Polycomblike proteins that include the mouse M96 and Drosophila Polycomblike. A partial cDNA for a human homolog of Tctex3, HUTEX3, has also been isolated. Mouse Tctex3 gene was mapped adjacent to Tsc2 gene on mouse Chromosome (Chr) 17, and HUTEX3 was located closely to HSET gene in the HLA class II region of chromosome 6. 相似文献
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Immunohistochemical localization of androgen receptor in mouse testicular germ cells during fetal and postnatal development 总被引:1,自引:0,他引:1
BACKGROUND: Determination of the cellular distribution of the androgen receptor (AR) in testicular cells is necessary for understanding the mode of AR action in the testis. We here investigated immunohistochemically the localization of AR by use of anti-human AR polyclonal antibody NH27, with special reference to the AR in germ cells in the developing mouse testis. METHODS: ICR mouse testes taken from day 14 post coitum (p.c.) to day 56 post partum (p.p) were used for AR immunohistochemistry by the routine immunoperoxidase method at the light microscopic level and the pre-embedding method at the electron microscopic level. RESULTS: On day 14 p.c., AR immunoreactivity was present in nuclei of prospermatogonia but not in those of Sertoli cells or interstitial cells. On day 14 p.p., the AR was detected in the nuclei of spermatogonia, Sertoli cells, and myoid cells. AR immunoreactivity in nuclei of Leydig cells appeared on day 21 p.p. In the mature mouse testis, the AR was present in the nuclei of spermatogonia, Sertoli cells, myoid cells, and Leydig cells. CONCLUSIONS: AR was present both in germ cells and in somatic cells during fetal and postnatal development of the mouse testis. In the fetal testis, AR was localized exclusively in prospermatogonia and spermatogonia, suggesting that androgen may act directly on germ cells during prespermatogenesis and the early stage of spermatogenesis. Based on the fact that AR is expressed in Sertoli cells, myoid cells, and Leydig cells around the onset of spermatogenesis, the regulation of AR expression in the germ cells seems to be different from that in the somatic cells. Furthermore, our present data suggest the ultrastructural localization in nuclei of mouse testicular cells is similar to that of some other steroid receptors, both in germ cells and somatic cells. 相似文献
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Y Matsui 《Canadian Metallurgical Quarterly》1998,42(7):1037-1042
The specification of mouse germ cell lineage takes place after a population of pluripotential cells is established, and cell communication among the pluripotential cells may be important for this process. Primordial germ cells (PGCs) first appear around the allantois at 7 dpc which are distinct from pluripotential cells in the early embryo because they can not colonize blastocysts. However, a portion of PGCs are transformed into pluripotential cells in the ectopic environment or in culture, suggesting that the developmental fate of PGCs may still be somewhat plastic. PGCs may be destined only for gametes after they enter into the mitotic arrest phase or the meiotic prophase in embryonic gonads, which may be regulated by intrinsic and/or environmental molecules. After fetal germ cells are mitotically arrested, a large number of germ cells undergo programmed cell death. Bcl-2 and its related molecules are involved in the determination of death or survival of fetal germ cells, as well as of spermatogonia in adult testis. The cell death of spermatogonia may be necessary either for eliminating impaired germ cells or for arranging optimal interactions between germ cells and their supporting cells. Although maturating germ cells seem to differentiate only to sperm cells, oocytes that complete the first meiotic division can give rise to pluripotential cells, suggesting that maternal molecules accumulated in oocyte may play a role in the restoration of pluripotency. 相似文献
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J Blanco-Rodríguez 《Canadian Metallurgical Quarterly》1998,21(5):236-248
The significance of cell death occurring during spermatogenesis is a subject of interest because of its potential medical importance. Unfortunately, the field has been difficult for andrologists to penetrate, in part because of the difficulties of studying germ cells in vitro and the complexity of designing suitable models in which to dissect the molecular signalling pathways involved in control of germ cell apoptosis. As a result, the reasons for these deaths remain unclear despite considerable investigative effort. As developments which have occurred over the last few years in understanding of apoptosis can shed light on this important topic, this review focuses on what is currently known about germ cell apoptosis and outlines the emerging picture of what might be the causes and biological role of germ cell deaths in spermatogenesis. 相似文献
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