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Endoreplication in the ovary, testis, and intestine of Strongyloides stercoralis.

During development of the free-living adults of the human parasitic nematode Stronglyoides stercoralis, cells in certain tissues grow by endoreplication in which rounds of DNA replication occur without cell or nuclear division. The DNA content of individual nuclei was measured by microdensitometry of Feulgen-stained preparations. In females, some ovarian cells have up to 800 times the haploid DNA content (800C). In males, some cells of the testis have up to 100C. Intestinal cells in both sexes have up to 16C, whereas most other somatic cells have 2C.

Animals↗

Interleukin-18 is expressed in rat testis and may promote germ cell growth.

Although host-defence mechanisms, designed to preserve the integrity of the developing germ cells are operative in the testis, the components of this protective system have yet to be characterised in detail. Here, we report that the cytokine interleukin-18 (IL-18) is expressed in the rat testis and may contribute to these defences. Thus, analysis by RT-PCR and Western blotting revealed pronounced testicular expression of pro-IL-18 from postnatal day 5 and onwards. Expression of both IL-18 mRNA and protein was found to be localised to meiotic and post-meiotic germ cells as evaluated by in situ hybridisation and immunohistochemistry, respectively. The mRNA species coding for the IL-18 receptor and IL-1beta converting enzyme, which activates pro-IL-18, were also shown to be expressed by the seminiferous tubules. Recombinant IL-18 was seen to stimulate spermatogonial DNA synthesis in cultures of staged segments of rat seminiferous tubules, without influencing germ cell apoptosis. These results suggest that IL-18 may have host-protective and growth-promoting functions in the testis, but further investigations need to be done to confirm this.

Animals↗

RIBONUCLEIC ACID SYNTHESIS DURING MITOSIS AND MEIOSIS IN THE MOUSE TESTIS.

The pattern of ribonucleic acid synthesis during germ cell development, from the stem cell to the mature spermatid, was studied in the mouse testis, by using uridine-H(3) or cytidine-H(3) labeling and autoradiography. Incorporation of tritiated precursors into the RNA occurs in spermatogonia, resting primary spermatocytes (RPS), throughout the second half of pachytene stage up to early diplotene, and in the Sertoli cells. Cells in leptotene, zygotene, and in the first half of pachytene stage do not synthesize RNA. No RNA synthesis was detected in meiotic stages later than diplotene, with the exception of a very low rate of incorporation in a fraction of secondary spermatocytes and very early spermatids. At long intervals after administration of the tracer, as labeled cells develop to more mature stages, late stages of spermatogenesis also become labeled. The last structures to become labeled are the residual bodies of Regaud. Thus, the RNA synthesized during the active meiotic stages is partially retained within the cell during further development. The rate of RNA synthesis declines gradually with the maturation from type A to intermediate to type B spermatogonia and to resting primary spermatocytes. "Dormant" type A spermatogonia synthesize little or no RNA. The incorporation of RNA precursors occurs exclusively within the nucleus: at later postinjection intervals the cytoplasm also becomes labeled. In spermatogonia all mitotic stages, except metaphase and anaphase, were shown to incorporate uridine-H(3). RNA synthesis is then a continuous process throughout the cell division cycle in spermatogonia (generation time about 30 hours), and stops only for a very short interval (1 hour) during metaphase and anaphase.

Animals↗

[Testicular metastasis of prostatic adenocarcinoma: report of 2 cases].

Prostatic carcinoma metastasizing to the testis is an extremely rare occurrence. In this study, two new cases have been reported of unilateral testicular metastases from an adenocarcinoma of the prostate. Unlike primary tumors of the testis, this type of tumor develops later in life (between 50 and 60 years of age), and is in most instances unilateral. It is generally discovered accidentally during orchiectomy or autopsy. It is not always easy to establish a morphological diagnosis, as the macroscopic appearance of the tumor may be normal. However, microscopic investigation reveals the presence of neoplasms of the testis.

Adenocarcinoma↗

Relaxin-like factor expression as a marker of differentiation in the mouse testis and ovary.

Expression of the relaxin-like factor (RLF) was studied at the messenger RNA (mRNA) and protein levels in the testes and ovaries of the mouse, as well as through testicular development and differentiation in the mouse testis. In situ hybridization or RT-PCR, and immunohistochemistry using a polyclonal antibody raised against a recombinant protein, provided mutually confirmatory results for a high expression of RLF in the Leydig cells of the adult testis and at a much lower level of expression in the luteal cells of the ovary through the cycle, pregnancy, and in lactation. Analysis of protein and mRNA expression, through postnatal testicular development, indicated moderate RLF expression also in the fetal population of Leydig cells, even in the hpg mutant mouse, lacking an active pituitary-gonadal axis. Prepubertal Leydig cells, however, exhibit only very low-level RLF gene expression, this phenotype persisting in the adult hpg mouse. In summary, fetal Leydig cells express RLF in an LH/human CG-independent fashion, whereas LH/human CG is essential to induce RLF expression in the adult-type Leydig cell. In cultured adult Leydig cells or in the mouse tumor MA-10 cell line, RLF mRNA is expressed in a constitutive fashion. RLF thus seems to be a useful marker of Leydig cell differentiation status.

Animals↗

Delay of testicular differentiation in the B6.YDOM ovotestis demonstrated by immunocytochemical staining for müllerian inhibiting substance.

It has been found that when the Y chromosome from Mus musculus domesticus (YDOM) is placed onto the C57BL/6J (B6) mouse background, the XY progeny (B6.YDOM) develop ovaries or ovotestes but not normal testes during fetal life. We examined the ontogeny of the abnormal testicular differentiation in the B6.YDOM ovotestis by immunocytochemical staining for Müllerian inhibiting substance (MIS). We found that the B6.YDOM ovotestis initiated testicular differentiation later in development than did the control B6 testis. When the YDOM was transferred onto the SJL J mouse background by crossing B6.YDOM males with SJL/J females, all XY progeny developed normal testes. The onset of testicular differentiation was at the same developmental stage as in the B6 male fetus. These results suggest that the delay of testicular differentiation is not due to the effect of the YDOM chromosome itself, but due to improper interaction of the testis-determining gene on the YDOM chromosome with autosomal genes of B6. In addition, we found a close correlation between the arrest of germ cells at the prespermatogonia stage and MIS production of adjacent somatic cells in the B6.YDOM ovotestis. This result may support the hypothesis that MIS is involved in the regulation of germ cell differentiation.

Animals↗

Effect of testosterone on development of the lumen in seminiferous tubules of the rat.

Although systemically administered testosterone can effect the postnatal maturation of elements in the seminiferous tubule in hypophysectomized rats and mice, it does not elicit the same degree of development which occurs in normal control animals. In view of reports of precocious spermatogenesis in androgen secreting Leydig cell tumors, the present study was designed to determine if high local levels of testosterone accelerate development of the seminiferous tubules. Testosterone pellets were inserted under the tunica albuginea of the right testis of 7 day old rats. At 17, 23 and 28 days of age development of the seminiferous tubules, as judged by the formation of tubule lumens, was more extensive in the treated testes than in contralateral and sham operated controls. Tubule diameters were not necessarily correlated with lumen formation. This study demonstrates that high local levels of testosterone accelerate seminiferous tubule development in the rat and indicates that tubule diameter may not be a valid basis for estimating development of the testis. It is suggested that testosterone exerts this effect through its actions on the Sertoli cells.

Aging↗

Differential expression of protein kinase C alpha and delta in testes of mouse at various stages of development.

Protein kinase C (PKC) describes a family of serine/threonine protein kinases, and multiple isoforms are expressed in various mammalian tissues. In the present study, we examined the expression of PKC-alpha and PKC-delta at protein and mRNA level in mouse testis by Western blotting and RT-PCR. We also examined the expression of both PKC isoenzymes in the developing mouse testis. In testes of mouse at various developmental stages, both the protein and the mRNA of PKC-alpha were uniformly distributed; but PKC-delta expression occurred in the testes of 3-week-old mice, perhaps even at a relatively late stage in spermatid development. The results suggest that each isoenzyme may have different functional roles in processing and modulating physiological cellular responses of spermatogenesis.

Aging↗

Germ cell apoptosis in the human testis.

Apoptosis is a widespread phenomena during development. It represents a form of cell death and has a crucial role in tissue homeostasis. Apoptosis is also involved in a number of pathological conditions. Spermatogenesis is a dynamic process of germ cell proliferation and differentiation. During regular spermatogenesis, the number of testicular germ cells degenerate by an apoptotic process. The significance of regulating cell population by apoptosis is more apparent when sperm production is halted. The presence and frequency of apoptosis in germ cells of human testis biopsy specimens were tested. The results confirm the presence of germ cell apoptosis but not the apoptosis of Sertoli cells. The increased apoptotic index was observed in patients with azoospermia in comparison with normal but obstructed spermatogenesis.

Apoptosis↗

Treatment of stage I testis seminoma by radiotherapy: long-term results--a 30-year experience.

PURPOSE: From 1956 to 1986, we have retrospectively studied 184 patients with a Stage I testis seminoma treated by orchidectomy and radiotherapy at the Institut Gustave Roussy. METHODS AND MATERIALS: The 184 patients received adjuvant radiotherapy to the para-aortic and ipsilateral iliac nodes. Of the 184 patients, 133 received additional mediastinal and supraclavicular irradiation, 47 received supraclavicular without mediastinum irradiation, 98 patients received additional radiotherapy given to inguino-scrotal area. The mean dose of irradiation is 21 Gy which is the lowest dose published. The actuarial survival rate is, respectively, 96%, 93%, 83% and 77% at 5, 10, 15, and 20 years. RESULTS: Four patients relapsed, and four died of progressive disease. Four patients presented cardiovascular disease, all of them had mediastinal irradiation, two were heavy smokers. Seventeen second malignancies were observed, six tumors in the contralateral testis. The actuarial risk of developing a second malignancy is 10% at 10 years, 21% at 20 years. The cure rate and relapse rate in our patients is the same as that obtained by higher dosage of irradiation. CONCLUSION: We conclude that low dose of prophylactic irradiation in lumbo aortic and ipsilateral iliac lymph nodes is active and safe in the treatment of Stage I testis seminoma.

Adolescent↗

[Fertility after orchiopexy for unilateral retention testis].

The testicular function after orchiopexy for unilateral cryptorchism was examined for fertile state with the relation to the age at operation and the testicular size. Sperm concentration decreased with the age at operation, accompanying a rise of gonadotropins and a decrease of the testicular size. The testicular size decreased with the age at operation. sperm deterioration was correlated more with the testicular size in the normally descending side than that of the operated one. The gonadotropin level was inversely correlated with the size of the contralateral normally descending testis. These results indicate that fertility after unilateral orchiopexy is influenced by the age at operation as well as the development of the normally descending contralateral testis.

Adolescent↗

Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique.

The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84.4%. Genes encoding cytochrome c oxidase II and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting their roles involved in meiosis.

Cloning, Molecular↗

Testicular tumours in maldescended testes.

Maldescent is known to increase the risk of cancer of the testis. However, the effect of orchiopexy on reducing tumour risk and altering tumour type or stage at diagnosis is not established. The authors review the records of patients with testicular tumours seen for seminoma (1958 to 1975) and nonseminoma (1958 to 1970). Of 646 patients, 53 (8.2%) had a history of testicular maldescent; in 42 (79%) the maldescent was unilateral and in 6 of these, the tumour developed in the normally descended testis. Twenty patients had successful orchiopexy at a median age of 14 years (range from 6 to 35 years). Tumours found in the 53 patients were seminoma (38 patients), teratoma (7 patients), embryonal carcinoma (6 patients), teratocarcinoma (1 patient) and choriocarcinoma (1 patient). Average age at the time of diagnosis of seminoma was 38 years and of nonseminoma 31 years. Average age was lower at the time of diagnosis of the tumour for those who had successful orchiopexy than for those who did not. Earlier stage nonseminomas were found in patients who had an orchiopexy. Twenty-five (66%) of the 38 seminomas occurred in testes located outside the scrotum, whereas only 6 (40%) of 15 nonseminomas occurred in a nonscotal position, suggesting that persistent maldescent favours seminoma over nonseminoma. In this retrospective review, no statement can be made about the effect of orchiopexy on tumour risk. However, this procedure appears to lead to an earlier diagnosis and may influence the type of tumour that subsequently develops.

Adolescent↗

Germ cell deficiency causes testis cord differentiation in reconstituted mouse fetal ovaries.

Sex-reversal in fetal ovaries was studied by using a dissociation-reconstitution technique. Gonads of 12.5 gestation-day male and female mouse fetuses were dissociated into single cells. To eliminate germ cells, the dissociated cells were cultured for 14 h, and then somatic cells attached to culture dishes were harvested and aggregated by gyratory culture for 24 h. The aggregates were then transplanted into ovarian bursa in ovary-ectomized nude mice. The recovered explants were examined histologically. Male somatic cells developed into testes containing Sertoli cells, Leidig cells, and tunica albuginea. Female somatic cells formed testis cords and differentiated into Sertoli cells, but they did not differentiate into other testis components or ovarian tissues. However, aggregates consisting of both female and male somatic cells differentiated into well-developed testes containing Leidig cells and tunica albuginea as well as Sertoli cells. Enzyme marker analysis showed significant contributions of female cells in these organized testes. In contrast, aggregates containing both female germ cells and somatic cells developed into ovaries and did not differentiate into any testicular tissues. The results indicate that female somatic cells in fetal gonads at 12.5 gestation day have the potency to form testis cords and differentiate into Sertoli cells. The subsequent steps in testis development require the contributions of male cells. The present study also suggests that testicular differentiation is independent of germ cells but ovarian development involves the interaction between germ cells and somatic cells.

Animals↗

An equine intersex with unilateral gonadal agenesis.

Cytogenetic and histological studies have been carried out on an intersex horse which was clinically diagnosed as a cryptorchid. The horse had the general conformation of a stallion but the external genitalia included a well developed vulva and a penis. The right testis which was descended was devoid of germ cells and the left "gonad" located in the cavum vaginale contained neither testicular nor ovarian tissue. The male duct system on both sides were relatively well developed despite the absence of a testis on the left side. Chromosome analysis on cultured cells from the descended testis revealed the presence of four chromosomally-distinct cell types with XX, XY, XXY and XO sex complements indicating a quadruple mosaicism. The presence of polymorphonuclear neutrophils exhibiting a drumstick, in the hemopoietic tissues and a sex chromatin body in the nucleated cells of buccal mucosa suggest that mosaicism prevails in other somatic tissues of the horse. On the basis of information derived from similar conditions in humans and some domestic animals it would appear that this horse resulted from an XXY zygote. The four cell types noted in the horse probably resulted through mitotic mechanisms favouring the loss of an X and a Y at different stages during embryonic development. The absence of gonad on the left side of this horse might be causally related to the preponderance of XO cell types in the somatic blastema during early gonadal differentiation.

Animals↗

Transcription factor GATA-4 is expressed in a sexually dimorphic pattern during mouse gonadal development and is a potent activator of the Müllerian inhibiting substance promoter.

Mammalian gonadal development and sexual differentiation are complex processes that require the coordinated expression of a specific set of genes in a strict spatiotemporal manner. Although some of these genes have been identified, the molecular pathways, including transcription factors, that are critical for the early events of lineage commitment and sexual dimorphism, remain poorly understood. GATA-4, a member of the GATA family of transcription factors, is present in the gonads and may be a regulator of gonadal gene expression. We have analyzed the ontogeny of gonadal GATA-4 expression by immunohistochemistry. GATA-4 protein was detected as early as embryonic day 11.5 in the primitive gonads of both XX and XY mouse embryos. In both sexes, GATA-4 specifically marked the developing somatic cell lineages (Sertoli in testis and granulosa in ovary) but not primordial germ cells. Interestingly, abundant GATA-4 expression was maintained in Sertoli cells throughout embryonic development but was markedly down-regulated shortly after the histological differentiation of the ovary on embryonic day 13.5. This pattern of expression suggested that GATA-4 might be involved in early gonadal development and possibly sexual dimorphism. Consistent with this hypothesis, we found that the Müllerian inhibiting substance promoter which harbors a conserved GATA element is a downstream target for GATA-4. Thus, transcription factor GATA-4 may be a new factor in the cascade of regulators that control gonadal development and sex differentiation in mammals.

Animals↗

Sex-specific fetal lung development and müllerian inhibiting substance.

Male neonates develop respiratory distress syndrome (RDS) with a greater incidence and mortality than do female neonates; the cause of this male disadvantage remains obscure. Male fetuses are exposed to higher levels of androgens and Müllerian inhibiting substance (MIS). Androgens have been shown to inhibit fetal lung maturation, and recent evidence in vitro indicates that MIS, a Sertoli cell-derived glycoprotein made early in ontogeny of the testis, may also inhibit lung development. To study whether this fetal regressor might inhibit maturation of the fetal lung in vivo, we injected human recombinant MIS (rMIS) into fetal rats, measured serum levels of rMIS using an enzyme-linked immunosorbent assay, and analyzed fetal lung tissue histologically and for protein, glycogen, DNA, and disaturated phosphatidylcholine content. Peak serum levels of recombinant MIS were measured at 6 h, with an apparent elimination half-life of 3 h, and without leakage into adjacent littermates injected with vehicle alone. Female fetal rat lung tissue exposed to recombinant MIS (10(-9) M, 10(-8) M) revealed depressed disaturated phosphatidylcholine content both 48 and 72 h after injection compared with female vehicle-injected littermates. Male lungs of the same gestational age appeared inhibited at a higher (10(-8) M) rMIS dose. These inhibitory effects observed in vivo confirm those previously seen in vitro and suggest that MIS, as well as androgens, may play a causative or important ancillary role in the sexual dimorphism that characterizes the neonatal respiratory distress syndrome.

Animals↗