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Photomanipulation of sexual maturation and breeding cycle of the steppe polecat (Mustela eversmanni) and other techniques for more rapid propagation of the species.

Twenty steppe polecats were divided into 2 groups, each consisting of 4 males and 6 females, and subjected to either a natural photoperiod (controls) or alternating periods of short (8 h light/16 hr dark for 8-9 weeks) and long days (16 h light/8 h dark for 16-20 weeks). The experimental photoperiod significantly accelerated sexual maturation in both sexes, with males developing maximal testis size within 57 days and females breeding after an average of 52 days exposure to 16L/8D. Males in the experimental group completed 2 1/2 testicular cycles and participated in mating during 3 successive breeding seasons during the 18 month period whereas males in the control group completed a single testicular cycle and only had an opportunity to mate during a single breeding season. Females in the experimental group produced 3 litters whereas females in the control group only gave birth to a single litter. Litter size averaged 6.9 +/- 2.0 (n = 23) and did not significantly differ with age, parity, or treatment. Pseudopregnant females returned to estrus within 12 days after the expected date of parturition, were bred, and gave birth to kits. Polecats which were subjected to the experimental photoperiods completed more molting cycles and underwent more photoperiod-induced changes in body weight than those in the control group. Death or removal of kits within 8 days after birth resulted in 12/12 females returning to estrus within 6-26 days. Eleven of these females were remated and gave birth to kits. Eight domestic ferrets readily accepted neonatal polecat kits and 5 successfully reared kits, although kit survival was quite poor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The influence of epididymal agenesis on the development and maturation of the testis: experimental model and clinical correlations.

The August X Copenhagen (ACI) rat provides an excellent model to study the effect of congenital epididymal anomalies on testicular function. Despite epididymal absence, testicular maturation and function are unchanged until puberty. At puberty, with increased testicular fluid and spermatozoa output, involution of germinal epithelium occurs. This involution is similar to the changes seen with ligation of the efferent ductules of the epididymis. These findings suggest that in this mammalian model the testis functions normally until the pubertal period. At that time, because of proximal epididymal agenesis and failure to reabsorb testicular secretions, intratesticular hydrostatic pressure increases with rapid germinal epithelial atrophy. Clinical correlations are made in patients with epididymal abnormalities, especially in association with undescended testis.

Animals↗

Testicular effects of the Leydig cell toxicant ethane dimethanesulphonate given to neonatal rats.

Neonatal rats were injected with either 50 mg/kg ethane dimethanesulphonate (EDS) or vehicle on days 1 to 5 inclusive or on day 1 alone. Studies were made on days 6, 28, and 63 of testicular structure; related endocrinologic parameters were measured in the day 1 to 5 treated animals only. Leydig cells and their activities were identified by cell counts using sections stained for 3 beta-hydroxysteroid dehydrogenase, hCG binding to LH receptors in testicular homogenates, and assays of intratesticular testosterone, plus pituitary and/or serum concentrations of testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH). Given on days 1 to 5, EDS reduced Leydig cell populations estimated by morphometry and 125I-HCG binding, and testicular and body weights between days 6 and 63, and permanently retarded the development of the seminiferous epithelium. Decreases of serum and intratesticular testosterone occurred with homeostatic rises in FSH and LH. Injection on day 1 reduced Leydig cell numbers only on day 6 although body weight remained retarded. The data illustrate the susceptibility of the developing rat testis to the cytotoxicant EDS; whether this is related to withdrawal of androgen production or nonspecific cytotoxicity remains to be evaluated.

Animals↗

Detection of endocrine disrupters: evaluation of a Fish Sexual Development Test (FSDT).

Managed by the Organisation for Economic Co-operation and Development (OECD), a comprehensive work is carried out in numerous laboratories to develop test guidelines for the detection of endocrine disrupting chemicals in humans, and various animal species. Development of tests to detect chemicals with endocrine disrupting properties in fish is a part of that work. A Fish Sexual Development Test (FSDT) (an extension of the existing OECD TG 210, fish early life stage toxicity test), proposed as an international test guideline for the detection of endocrine disrupting chemicals, was evaluated by water exposure of juvenile zebrafish to the three natural estrogens: estrone, 17beta-estradiol, and estriol and the synthetic androgen trenbolone (trenbolone acetate). As endpoints, vitellogenin induction and histological changes including changes in sex ratios were investigated. The sex ratio was significantly altered towards females from 49 ng/l estrone, 54 ng/l 17beta-estradiol and 22 microg/l estriol, respectively. An all male population was observed from exposure to 9.7 ng/l trenbolone and above. Significant vitellogenin induction in whole body homogenate was measured after exposure to 14 ng/l estrone, 54 ng/l 17beta-estradiol and 0.6 mug/l estriol, respectively. Significant vitellogenin reduction was measured after exposure to 193 ng/l trenbolone or higher. The present results provide strong evidence that the FSDT is a sensitive test toward estrogenic and especially androgenic exposure and the validation of the FSDT as an OECD test guideline should continue.

Animals↗

Effects of the fungicide prochloraz on the sexual development of zebrafish (Danio rerio).

Some chemicals have the potential to adversely affect sexual development through multiple endocrine actions. Prochloraz is an imidazole fungicide that displays diverse mechanisms of action, including inhibition of aromatase activity, inhibition of androgen synthesis, and antagonism of the androgen receptor. The objective of this study was to assess the effects of prochloraz on the sexual development of zebrafish (Danio rerio) in the Fish Sexual Development Test (FSDT) proposed as an OECD test guideline for detection of endocrine disruptors. Zebrafish were exposed to prochloraz (0, 16, 64 or 202 microg/L) for 60 days from 24 h post fertilization. Fish exposed to 202 microg/L prochloraz showed an increased proportion of males. Furthermore, the incidence of intersex and the stages of the gonads were altered in the treated fish compared to the control fish. A significant vitellogenin decrease was observed in both female and male zebrafish at an exposure concentration of 202 microg/L prochloraz. However, in the male fish, significantly increased vitellogenin concentrations were observed in the groups exposed to 16 or 64 microg/L prochloraz. This study serves as a part of the validation of the FSDT and indicates that the FSDT is suitable in detecting compounds with multiple endocrine actions. This is of importance in the assessment of the potential risk of existing and new chemicals.

Animals↗

Life-stage-dependent sensitivity of zebrafish (Danio rerio) to estrogen exposure.

The aim of this study was to identify periods in zebrafish (Danio rerio) development when estrogen exposure has long-term consequences on reproductive capabilities at the adult stage. To this end, zebrafish were exposed to 10 ng/L ethynylestradiol (EE(2)) during three stages of gonadal differentiation: (i) the juvenile hermaphroditic stage when gonads display the morphology of an immature ovary (in our zebrafish colony this lasted from 15 to 42 days post-fertilization [dpf]), (ii) the gonad transition stage when the hermaphroditic gonad differentiates into either testes or ovary (from 43 to day 71 dpf), and (iii) the premature stage of testicular and ovarian development (from 72 to 99 dpf). The consequences of stage-specific exposure to EE(2) were assessed by determining time to first spawning, fecundity (number of eggs per female per day), fertilization success (percentage of fertilized eggs) and sex ratio of the adults. Exposure during the gonad transition period induced a delay in the onset of spawning and a significant reduction of fecundity and fertilization success, whereas exposure during the hermaphroditic stage or during the premature stage had no significant impact on the reproductive parameters of adult fish. The results from this experiment pointed to the gonad transition stage as being most susceptible to persistent effects of developmental estrogen exposure. In a second experiment, the concentration dependency of the EE(2)effects was evaluated by exposing zebrafish during the gonad transition stage (43-71 dpf) to 1.67, 3 or 10 ng EE(2)/L. Significant effects of EE(2) on adult reproduction were found with 3 and 10 ng EE(2)/L, but not with 1.67 ng/L. Histological examination of the gonads revealed that at termination of EE(2) exposure (71 dpf), all individuals in the 3 and 10 ng EE(2)/L treatment possessed ovaries. However, this feminising effect appeared to be reversible since at the adult stage (190 dpf), both fish with ovaries and with testes were found. Thus, EE(2) exposure during the gonad transition stage seems to have no persistent effect on gonad histology but on reproductive capabilities.

Animals↗

Effect of nutrition during calfhood and peripubertal period on serum metabolic hormones, gonadotropins and testosterone concentrations, and on sexual development in bulls.

The objective of the present study was to characterize the effects of nutrition on circulating concentrations of metabolic hormones, gonadotropins, and testosterone during sexual development in bulls. Nutrition regulated the hypothalamus-pituitary-testes axis through effects on the GnRH pulse generator in the hypothalamus and through direct effects on the testes. Pituitary function (gonadotropin secretion after GnRH challenge) was not affected by nutrition. However, nutrition affected LH pulse frequency and basal LH concentration during the early gonadotropin rise (10-26 weeks of age). There were close temporal associations between changes in insulin-like growth factor-I (IGF-I) concentrations and changes in LH pulse frequency, suggesting a role for IGF-I in regulating the early gonadotropin rise in bulls. The peripubertal increase in testosterone concentration was delayed in bulls with lesser serum IGF-I concentrations (low nutrition), suggesting a role for IGF-I in regulating Leydig cell function. Serum IGF-I concentrations accounted for 72 and 67% of the variation in scrotal circumference and paired-testes volume, respectively (at any given age), indicating that IGF-I may regulate testicular growth. Bulls with a more sustained elevated LH pulse frequency during the early gonadotropin rise (high nutrition) had greater testicular mass at 70 weeks of age relative to the control group (medium nutrition), despite no differences in metabolic hormone concentrations after 26 weeks of age. Therefore, gonadotropin-independent mechanism regulating testicular growth might be dependent on previous gonadotropin milieu.

Animals↗

Sperm parameters after early left varicocele treatment.

OBJECTIVE: We evaluated whether early treatment for left varicocele can improve sperm parameters. DESIGN: Patient follow-up and comparison with control groups. SETTING: Surgical department of a Children's Hospital and a university infertility-care center. PATIENT(S): Nineteen patients who underwent surgery for left varicocele as adolescents (mean +/- SD, 14.11 +/- 1.59 years) were followed up as adults (mean +/- SD, 19.37 +/- 1.16 years). MAIN OUTCOME MEASURE(S): A testicular examination was performed and varicocele staging was done at the time of surgery. Andrologic examination and semen analysis were done 2-8 years later. Semen results were compared with those of two age-matched control groups of 19 healthy, randomly selected subjects and 19 unoperated patients suffering from left varicocele of the same degree. RESULT(S): The mean values of sperm parameters in the patient study group were similar to those of the healthy controls except that the sperm concentration was significantly lower. However, sperm motility and morphology mean values in the patient study group were significantly higher than those of the unoperated patients. CONCLUSION(S): This study indicates that surgery in adolescence can be useful, possibly because the testis is developing at that age. Large-scale, double-blind controlled trials on early treatment of left varicocele are needed to identify possible markers of disease severity.

Adolescent↗

Gonadal differentiation: a review of the physiological process and influencing factors based on recent experimental evidence.

The sexual differentiation of a gonad is determined in normal embryology by the presence or absence of the TDF gene, a short segment of DNA localized near the tip of the short arm of the Y chromosome. Under the stimulus of this gene, the somatic cells of the genital ridge differentiate into Sertoli cells and secrete anti-müllerian hormone. This hormone inhibits the germ cells from entering meiosis and may well also trigger the formation of the primary sex cords. Once the Sertoli cells are formed, further differentiation of the gonad is independent of the TDF presence, although this gene may further contribute to the normal functioning of the male gonad. In true hermaphroditism the normal regulation of the gonadal differentiation is missing. Gonadal differentiation may revert to a more primitive evolutionary level at which the rate of growth of the gonad determines its sexual direction. When this exceeds a certain threshold a testis may develop despite the absence of the TDF gene.

Animals↗

Effect of 17beta-estradiol on the reproduction of Japanese medaka (Oryzias latipes).

Estrogenic compounds such as 17beta-estradiol (E2) and its analogs are present in the aquatic environment and can adversely affect the reproductive systems of aquatic organisms. Although E2 has been detected at a maximum concentration of 64 ng/l in effluents of sewage treatment works (STWs), few reports address the associated effects on reproduction in fish. Therefore, we exposed adult medaka (Oryzias latipes) to mean measured E2 concentrations of 29.3, 55.7, 116, 227, and 463 ng/l for 21 d and assessed the effects on the egg number and fertility of paired medaka during the exposure period. In addition, we determined the hepatic vitellogenin (Vtg) concentration and histologically assessed the gonads of these fish. The number of egg produced and fertility of the paired medaka exposed to 463 ng/l E2 were significantly less compared with those of the control fish. Males in all treatment groups had developed testis-ova. Males treated with E2 concentrations = 55.7 ng/l contained relating great concentrations of hepatic Vtg. Therefore, although only the greatest E2 concentration tested in our study affected fecundity and fertility, effects of E2 were observed on induction of Vtg and testis-ova in male medaka exposed to environmentally relevant concentrations of E2.

Animals↗

Kaposi's sarcoma following chemotherapy for testicular cancer in a homosexual man: demonstration of cytomegalovirus RNA in sarcoma cells.

A 35-year-old white Jewish homosexual man who had undergone surgery and chemotherapy for an embryonal carcinoma of the testis subsequently developed Kaposi's sarcoma. The neoplasm involved the skin as well as visceral tissues. Tissue derived from a biopsy specimen of one of the skin lesions was used in the in situ hybridization technique for the detection of genetic material. Cytomegalovirus messenger RNA was identified in the neoplastic Kaposi cells in the skin. The significance of this finding is discussed.

Cytomegalovirus↗

Isolated testicular polyarteritis nodosa mimicking testicular neoplasm-long-term follow-up.

Polyarteritis nodosa is a systemic vasculitis characterized by segmental necrotizing lesions of medium and small-size arteries. Rarely, the inflammatory process is isolated and involves a single organ without systemic manifestations. We describe a patient with isolated polyarteritis nodosa of the testis who presented with a testicular mass mimicking primary testicular tumor. The postoperative laboratory evaluation was negative. Long-term follow-up, without systemic treatment, showed no evidence of recurrence in the remainder of the testis or development of systemic vasculitis.

Adult↗

Promoter characterization of the human and mouse epilysin (MMP-28) genes.

Epilysin (MMP-28) is a recently cloned member of the matrix metalloproteinase family (Lohi et al., J. Biol. Chem. 276 (2001) 10134). It is expressed at highest levels in the skin by basal and suprabasal keratinocytes, and in testis by developing germ cells. To characterize the epilysin promoter, we isolated a 3.0 kb fragment of human genomic DNA containing 5'-flanking sequence of the epilysin gene, and a corresponding 660 bp fragment from the mouse. The 5'-flanking sequences contain no typical TATA-boxes or CCAAT sequences close to the translation initiation sites. RNase protection assay revealed that two transcription start sites are utilized in the human epilysin gene, situated 210 and 230 bp upstream from the translation start site. The promoter contains a GT-box, situated 300 bp upstream from the translation start site, with homology to the consensus binding site for transcription factors of the Sp family. This site is perfectly conserved between the human and mouse promoters. For reporter gene assays a series of constructs with fragments of increasing length of the epilysin promoter were coupled to the firefly luciferase gene. Reporter gene assays indicated that deletion or mutation of the GT-box dramatically reduces the transcriptional activity both in keratinocytes and in spermatogonia. Gel mobility shift assays showed that several nuclear proteins bind specifically to this sequence. Supershift assays with antibodies specific for members of the Sp family identified Sp1 and Sp3 as components of these protein/DNA complexes and hence as possible regulators of the epilysin gene. Our results indicate that the epilysin promoter has distinctive structural and functional features, which may control the unique expression and regulation patterns of the epilysin gene.

Animals↗

Expression of an antigen associated with basal bodies of human ciliated epithelial cells.

The process and regulation of ciliogenesis in human epithelia is little understood and many components of the cilium and associated structures have not been characterised. We have identified a monoclonal antibody, LhS28, which recognises a 44,000-45,000 M(r) protein specifically associated with human ciliated epithelial cells. Immunoperoxidase labelling of formalin-fixed paraffin wax-embedded human tissues showed that LhS28 was expressed in the sub-apical zone of ciliated epithelial cells of the Fallopian tube and upper respiratory tract, but not ciliated ependyma, non-ciliated epithelia or testis containing developing spermatozoa. Immunoelectron microscopy demonstrated that the antigen recognised by LhS28 was associated with the basal body structure of the cilium and specifically with the 9 + 0 microtubule arrays. LhS28 should be a useful tool in the identification of ciliated cells in pathological specimens and for investigating mechanisms of ciliogenesis.

Antibodies, Monoclonal↗