Müllerian remnants of male mice exposed prenatally to diethylstilbestrol

Prenatal exposure of males to diethylstilbestrol (DES) results in reproductive tract teratogenesis, ie, retention of Müllerian duct remnants. The potential of these remnants to develop pathological changes has not been studied. Therefore, pregnant outbred CD-1 mice were subcutaneously injected with daily doses of DES (100 micrograms/kg) on days 9 through 16 of gestation. DES-exposed male offspring and age-matched control male mice were sacrificed at 10 to 18 mo of age and examined for reproductive tract abnormalities. Prominent Müllerian remnants were observed in 268 out of 277 (97%) of the DES-exposed male mice. These remnants differentiated into "femalelike structures" homologous to oviduct and uterus. The Müllerian remnants were often enlarged and cystic and shared supporting connective tissue with adjacent male structures. Previously reported lesions, termed "epididymal cysts," were determined histologically to be cystic "oviductlike" structures and were, therefore, considered a Müllerian duct abnormality. Pathological changes in these male oviductal and uterine homologs included benign and malignant lesions. In addition, epididymal structures were altered. Inflammation and sperm granulomas were prevalent in DES-treated mice as young as 10 mo old but were only observed in control mice at 18 mos. Cysts of epididymal duct origin, hyperplasia, and adenoma of the epididymal duct were also observed. No comparable abnormalities were noted in 122 control males of corresponding ages. The data presented in this report demonstrated that transplacental exposure to DES affected the differentiation and normal development of the male genital tract involving both the Müllerian (paramesonephric) and Wolffian (mesonephric) ducts. The long-term changes in these tissues include lesions, some of which resembled neoplasia although the natural history of the lesions is not known. Moreover, some previously described abnormalities referred to as "epididymal cysts" were associated with tissues derived from embryonic female origin.     

Accelerated differentiation in seminiferous tubules of fetal mice prenatally exposed to ethinyl estradiol

Summary Ethinyl estradiol (EE) in olive oil (0.02, 0.2, or 2.0 mg/kg) administered to pregnant mice on days 11 to 17 of pregnancy induced abnormal differentiation of gonocytes and fetal Sertoli cells in male fetuses on day 18 of gestation. Light and electron microscopic examination of the testes showed fewer darkly stained prospermatogonia and more lightly stained prospermatogonia in the experimental than in the control fetuses. Widespread degeneration and lysis of gonocytes were seen only in the experimental mice. No spermatogonia type A could be detected. In spite of comparable mitotic rates in the Sertoli cells of the experimental and control mice and more dark Sertoli cells with well developed smooth endoplasmic reticulum (SER) in the experimental mice, one of the functions of fetal Sertoli cells was suppressed: there were fewer dark slender Sertoli cells with long processes extending to the centers of tubules and more contact areas with gonocytes, phenomena which may play a role in the migration of gonocytes towards the periphery of the tubules. More Sertoli cells were detected in the undescended than the descended testes exposed to the highest dose of EE. These morphological findings indicate that prenatal exposure to EE induces acceleration of prespermatogenesis and disturbances in the initiation of spermatogenesis and in the mechanical function of Sertoli cells.Supported by a grant from Sankyo Foundation of Life Science     

Altered splenocyte function in aged C57BL/6 mice prenatally exposed to diethylstilbestrol

The linkage between in utero exposure to diethylstilbestrol (DES) and the manifestation of a variety of reproductive disorders and possibly immune alterations in adults (i.e., human and mice) is suggestive of a fetal basis of adult disease. While the long-term adverse consequences of prenatal DES-exposure on reproductive disorders are well known, there is a paucity of data with regard to immune outcome. We hypothesize that prenatal DES-exposure "imprints" the immune system, altering the response to subsequent exposure to DES in adult mice. In this pilot study, C57BL/6 mice were prenatally exposed to DES or vehicle only (oil) and then exposed to DES at 1 year of age. Potential alterations in the spleen were then examined. Female DES-exposed mice (DES(prenatal)/DES(adult)) or female(DES) had higher serum levels of interferon-gamma (IFNgamma) in response to administration of an IFNgamma -inducer (soluble proteins-derived from Toxoplasma gondii), compared to female controls, which received oil during prenatal life (Oil(prenatal)/DES(adult)). Splenic lymphocytes from female DES(prenatal)/DES(adult) mice, when activated with Concanavalin A (ConA), also secreted higher levels of IFNgamma compared to female controls (Oil(prenatal)/DES(adult)) when examined at 14-months of age. This increase in IFNgamma in prenatal DES-exposed mice is not due to enhanced numbers of splenocytes or increased relative percentages of CD4(+) or CD8(+) cells. ConA-activated T-cells from female DES(prenatal)/DES(adult) had increased expression of the co-stimulatory molecule, CD28. These above immune changes were not evident in the males prenatally exposed to DES. Prenatal DES exposure also did not induce autoimmunity in non-autoimmune C57BL/6 mice. Overall, results from these prefatory studies suggest that prenatal DES exposure may have long-term immune alterations, which become evident following a secondary exposure to DES in adult life.     

Behavior and serotonergic disorders in rats exposed prenatally to valproate: A model for autism

In order to explore whether some aspects of the autistic phenotype could be related to impairment of the serotonergic system, we chose an animal model which mimics a potential cause of autism, i.e. rats exposed to valproate (VPA) on the 9th embryonic day (E9). Previous studies have suggested that VPA exposure in rats at E9 caused a dramatic shift in the distribution of serotonergic neurons on postnatal day 50 (PND50). Behavioral studies have also been performed but on rats that were exposed to VPA later (E12.5). Our aim was to test whether VPA exposure at E9 induces comparable behavioral impairments than at E12.5 and causes serotonergic impairments which could be related to behavioral modifications. The results showed significant behavioral impairments such as a lower tendency to initiate social interactions and hyperlocomotor activity in juvenile male rats. The serotonin levels of these animals at PND50 were decreased (−46%) in the hippocampus, a structure involved in social behavior. This study suggests that VPA could have a direct or indirect action on the serotonergic system as early as the progenitor cell stage. Early embryonic exposure to VPA in rats provides a good model for several specific aspects of autism and should help to continue to explore pathophysiological hypotheses.     

Sexual activity level and sexual functioning in women prenatally exposed to diethylstilbestrol

Thirty women with a history of prenatal exposure to diethylstilbestrol (DES) underwent a detailed sexual history and were compared to a demographically similar sample of 30 women with a history of an abnormal Pap smear. The DES women were found to have less well-established sex-partner relationships and less experience with child-bearing, to be lower in sexual desire and enjoyment, sexual excitability, and orgasmic coital functioning, but to be comparable (and low) with regard to such sexual dysfunctions as vaginismus and dyspareunia. Both potential psychosocial and neuroendocrine explanations are discussed.     

孕期非那雄胺暴露致子代雄性小鼠生殖器官发育异常

目的:探讨孕期非那雄胺暴露对子代雄性小鼠生殖器官发育的影响。方法:CD-1小鼠在受孕后0～17 d给予非那雄胺处理,通过宏观观察、解剖分析与组织形态学染色观察子代雄性小鼠生殖器官的发育情况;通过免疫荧光染色分析子代雄性小鼠精子发生情况。结果:宏观观察结果显示,孕期非那雄胺暴露可导致子代雄性小鼠外生殖器官畸形,表现为阴囊未完全融合及阴茎畸形;此外,还观察到小鼠肛门与生殖器的距离显著缩短(P<0.01)。解剖分析结果显示,孕期非那雄胺暴露可导致子代雄性小鼠睾丸不同程度的不完全下降及长度显著缩短(P<0.01)。组织形态学结果显示,各阶段阴茎长度均显著缩短(P<0.01);睾丸生精小管密度和生精小管管腔成熟精子数均显著降低(P<0.01),生精小管管腔和睾丸间质间隙均显著增大(P<0.01)。免疫荧光染色结果显示,睾丸中支持细胞、睾丸间质细胞和精原细胞的密度均显著降低(P<0.01);生精小管细胞的caspase-3荧光强度显著增加(P<0.01),Ki67与沙漠刺猬因子(desert hedgehog,Dhh)荧光强度均显著降低(P<0.01...     

A quantitative study of the facial nerve in mice prenatally exposed to ethanol

ABSTRACT  Pregnant ICR mice were given 20 % ethanol intraperitoneally twice on day 13 of gestation and allowed to give birth to offspring. The offspring were killed at 56 days of age and the motor root of their facial nerve was examined histologically and morphometrically. The cross-sectional area of the facial nerve of mice prenatally exposed to ethanol was significantly smaller than that of the control mice. There was no significant difference in the total number of myelinated axons or the mean axonal diameter between control and ethanol-exposed mice, but the mean diameter of myelinated fibers (axon + myelin sheath) and the thickness of myelin sheath were significantly decreased in the treated group. These results suggest that prenatal exposure to ethanol disturbs myelina-tion of the motor root of the facial nerve and may cause permanent neurological effects.     

Sexual behavior, neuroendocrine, and neurochemical aspects in male rats exposed prenatally to stress

The present study was designed to examine some short- and long-term effects of maternal restraint stress--during the period of sexual brain differentiation--on reproductive and endocrine systems, sexual behavior, and brain neurotransmitters in male rat descendants. Pregnant rats were exposed to restraint stress for 1 h/day from gestational days (GDs) 18 to 22. Prenatal stress did not influence the wet weight of sexual organs and the quantity of germ cells in adult male pups; however, these animals showed reduced testosterone levels, delayed latency to the first mount and first intromission, and also decreased number of ejaculations. Additionally, there was an increase in the dopamine and serotonin levels in the striatum. Our results indicate that prenatal stress had a long-term effect on neurotransmitter levels and sexual behavior. In this sense, reproductive problems caused by injuries during the fetal period can compromise the later success of mating as well as the capacity to generate descendants.     

Behavioral outcome of preschoolers exposed prenatally to cocaine: role of maternal behavioral health

OBJECTIVE: To examine the impact of prenatal cocaine exposure and maternal behavioral health (recent drug use and psychological functioning) on child behavior at age 5 years. METHOD: In this longitudinal investigation, maternal report of child behavior was assessed using the Achenbach Child Behavior Checklist (CBCL) in 140 cocaine-exposed and 181 noncocaine-exposed (61 alcohol, tobacco, and/or marijuana-exposed, and 120 nondrug-exposed) low-income, African American children. Structural equation modeling was used to estimate suspected causal relationships between indicators of maternal behavioral health at 5-year follow-up, according to self-report on a modified Addiction Severity Index (ASI) and CBCL scores. RESULTS: Prenatal cocaine exposure was not related to child behavior at age 5. Recent maternal drug use and psychological functioning had relationships with CBCL Internalizing and Externalizing scores. However, when considered within a combined model, only maternal psychological functioning remained significant. CONCLUSIONS: Findings highlight the importance of maternal functioning in the behavioral outcome of children exposed prenatally to cocaine.     

Salivary cortisol levels relate to cognitive performance in children prenatally exposed to methadone or buprenorphine

Opioid maintenance therapy (OMT) is generally recommended for pregnant opioid-dependent women. However, much is still unknown about the potential long-term effects of prenatal methadone and buprenorphine exposure. This study explored the long-term effects of prenatal methadone and buprenorphine exposure in a cohort (n = 41) of children, aged 9–11 years, using the Wechsler Abbreviated Scale of Intelligence (WASI) to measure cognitive development and salivary cortisol samples to measure HPA-axis activity. Prenatally exposed children scored significantly lower on all four subtests of WASI (vocabulary, similarities, block design, and matrix reasoning), compared to a comparison group (all p < .05). No group differences were found for salivary cortisol levels or cortisol reactivity levels (all p > .05). Cortisol levels significantly predicted matrix reasoning scores for the OMT group, β = −65.58, t(20) = 15.70, p = .02. Findings suggest that prenatal exposure to methadone or buprenorphine does not have long-term effects on children's HPA-axis functioning. However, since children of women in OMT scored significantly lower on tasks of cognitive function, careful follow-up throughout the school years and across adolescence is recommended.     

Open-field behavior in rats exposed prenatally to a low intensity-low frequency, rotating magnetic field

Two experiments were conducted to study the behavioral effects of prenatal exposure to a low intensity, ultra-low-frequency magnetic field. In Experiment I, 117 albino rats that had been exposed continuously during their prenatal development to a 3 to 30 gauss, 0.5 Hz rotating magnetic field (RMF), and 83 control rats that had been exposed renatally to control conditions, were tested in an open field at 21 to 25 days of age. RMF-exposed animals traversed significantly fewer squares than their controls in the open field (p < .001), but defecated significantly more in that situation (p < .001). RMF-exposed males also traversed significantly fewer squares than the RMF-exposed females (p < .05). Three RMF-exposed litters that were nursed by control mothers did not differ significantly in open-field activity from the pups in the 4 RMF-exposed litters from which they were taken at birth. In Experiment 2, in which the experimenters did not know whether the subject was a RMF-exposed rat or a control rat, 19 RMF-exposed rats again traversed significantly fewer squares than the 20 control rats (p < .01).     

Hemoglobinemia in mice exposed to high altitude

Male and female mice were exposed to a simulated altitude of 5,500m for 10, 30 or 90 days. After exposure to altitude for 90 days one group of each sex was then returned to sea level pressures for 10 days. In addition to the expected increases in hematocrit and hemoglobin, altitude exposure increased the 2,3-DPG content of red cells. Maximum values of 2,3-DPG (moles/g hemoglobin) occurred after 10 days at altitude and thereafter declined to values comparable to those in sea level mice despite continued exposure to hypoxia.As previously described for rats, mice also exhibit an altitude-induced hemoglobinemia which occurs in the face of a polycythemia. Plasma hemoglobin concentrations were at a steady state after about 30 days at altitude. Both sexes also showed a significant splenic hypertrophy, a bilirubinemia and a hemoglobinuria as a result of altitude exposure. As assessed in male mice a significant induction of heme oxygenase activity occurred in the kidneys but not in the spleen or liver. The latter finding contrasts with results previously observed in rats where induction of heme oxygenase occurs in the liver desplte a decrease in the liver/body weight ratio.Sex differences in the response to hypoxia previously observed with rats also occur in mice. The altitude-induced rise in hematocrit but not hemoglobin appears to be more pronounced in males as was the failure to gain weight. Perhaps both male mice and rats are more severely compromised by altitude than are females of the same speciesThe time course of the hemoglobinemia and its reversal at sea level paralleled the changes in hemoglobin and hematocrit suggesting that it was correlated with the demand on hematopoietic effort. Perhaps the hemoglobinemia is secondary to ineffective hematopoiesis.     

淋巴滤泡中补体成分沉积的意义

目的:观察补体成分在淋巴组织中的表达,探讨不同发育阶段淋巴滤泡中补体激活与免疫球蛋白沉积及滤泡树突状细胞之间的关系。方法:采用免疫组化方法对淋巴组织标本进行IgG、IgM、IgA、C3c、C4c、C1q和C4d标记,同时对其部分标本进行双标染色。结果:淋巴组织次级滤泡的Ⅰ~Ⅳ期的生发中心才出现不同程度的IgG、IgM、IgA、C3c、C4c、C1q、C4d阳性表达;免疫球蛋白与补体成分重叠沉积,并呈不规则网线状包绕在滤泡树突状细胞周围。其中以C4d的阳性表达最强。结论:淋巴滤泡生发中心有大量免疫球蛋白和补体成分围绕滤泡树突状细胞沉积,可能与中心淋巴细胞的发育和选择有关。     

Effects of indomethacin on preovulatory follicles in immature,superovulated mice

In order to demonstrate the possible role of prostaglandins in preovulatory follicular development, immature mice superovulated with pregnant mare serum followed 40 hours later by luteinizing hormone (LH) were treated with the prostaglandin-synthetase inhibitor, indomethacin. Indomethacin (10 mg/kg) injected at varying intervals prior to or following LH inhibited ovulation most effectively when administered within 2 hours of the ovulatory gonadotropin. This inhibition was accompanied by (1) suppression of the morphological changes normally occurring within the follicular wall during preovulatory development and (2) failure of germinal vesicle breakdown (GVBD) in two-thirds of the follicles examined. When GVBD occurred, indomethacin treatment appeared to delay meiotic maturation. Cumulus tissue was more compact than in control follicles and maintained a close association with the oocyte. These results suggest that alterations in the morphology of the follicle prior to ovulation—specifically, thinning of the apical follicular wall and meiotic maturation—are regulated by prostaglandins.     

DJ-1缺失增加出生前脂多糖暴露引起的出生后小鼠多巴胺能神经元丢失(英文)

目的:观察DJ-1缺失和出生前脂多糖(lipopolysaccharide,LPS)暴露对出生后小鼠多巴胺能(dopam-inergic,DA)系统和神经炎症的影响。方法:妊娠第10.5 d DJ-1基因敲除的小鼠腹腔注射脂多糖(10,000 EU/kg体重)后自然分娩的后代在4月和14月龄时处死。免疫组织化学染色结合体视学计数定量黑质酪氨酸羟化酶(Tyrosine hydroxylase,TH)和CD11b阳性细胞数量。高效液相色谱法测定纹状体DA代谢物水平。免疫荧光法测定TNF-α和IL-1β蛋白水平。结果:4月和14月龄的DJ-1基因敲除的小鼠均没有明显的DA神经元减少和脑内神经炎症改变。出生前接触过LPS的4月和14月龄野生型小鼠,黑质DA能神经元分别减少13.6%和23.1%,纹状体DA含量分别减少19.0%和26.2%,同时神经炎症改变明显。出生前接触过LPS的4月和14月龄DJ-1基因敲除小鼠,黑质DA能神经元分别减少22.5%和35.4%,纹状体DA含量分别减少34.3%和39.3%,同时脑内炎症反应更明显。结论:以上结果提示遗传因素缺失和环境因素可能共同引发帕金森病发生,进一步验证了帕金森病多因素损伤引发的发病假说。     

Changes in Ca(2+)/calmodulin-dependent protein kinase II activity and its relation to performance in passive avoidance response and long-term potentiation formation in mice prenatally exposed to diethylstilbestrol

We investigated the effects of prenatal exposure to diethylstilbestrol (DES), an endocrine disrupter on learning behavior and synaptic functions. Specifically, we determined the activity of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and related kinases that play an essential role in long-term potentiation (LTP) in the hippocampus in mice that were prenatally exposed to DES. Treatment with DES resulted in increased CaMKII autophosphorylation and Ca(2+)-independent activity in the hippocampus and cortex of male mice. Impaired passive avoidance correlated with this increased CaMKII autophosphorylation, as did the enhanced early phase of LTP (E-LTP) in hippocampus. These data suggest that prenatal exposure to DES induces deficits in passive avoidance responses as a result of increased CaMKII activity and hippocampal LTP.     

Morphometric characteristics of the primordial to primary follicle transition in the human ovary in relation to age

BACKGROUND: The growth pattern of the smallest ovarian follicles in humans is still incompletely documented. Using follicle hemispheres in thick histological sections, morphometric changes of primordial to primary follicles and possible age-related effects were evaluated. METHODS: In ovarian sections from 25 females aged 4-39 years a total of 1122 primordial, transitory or primary follicles were assessed for the diameters of follicles, oocytes and oocyte nuclei and for number of granulosa cells (GCs). RESULTS: The number of GCs in primordial, transitory and primary follicles were approximately 30, 50 and 100, respectively. The diameters of primordial follicles and oocytes increased with the woman's age until the mid-30's, after which time they decreased in size. The number of GCs in primordial follicles showed a moderate increase with age, whereas the number of GCs in transitory follicles showed a clear increase with age . The oocyte nucleus diameter (14-23 microm) showed significant linear correlations with the oocyte and follicular diameters and number of GCs in the follicle, while the number of GCs in the whole follicle and in the largest cross-section were closely related to the oocyte diameter. CONCLUSIONS: The number of GCs in small follicles is accurately estimated and shows an increase with age, indicating that the starting point of follicular development alters with female age. The age-related changes observed may be linked to the poorer reproductive performance of older women.