邻苯二甲酸二丁酯诱导雄性大鼠发生尿道下裂的分子作用机制

目的探讨邻苯二甲酸二丁酯（DBP）孕晚期染毒诱导雄性大鼠发生尿道下裂的分子作用机制。方法雌鼠怀孕（GD）第14—18天，每天分别灌胃予大豆油，DBP500或每日800mg／kg体重，雄性仔鼠出生后（PND）第1天，鉴别出DBP染毒组中发生尿道下裂的雄性仔鼠，分组为雄性仔鼠对照组（A组），每日500mg／kg体重（H）（B组）和每日800mg／kg体重（H）（C组）。用实时定量逆转录-聚合酶链反应（RT-PCR）方法检测阴茎组织中Shh，骨形态发生蛋白4（Bmp4）和雄激素受体（Ar）mRNA表达水平。用Westernblot测量阴茎组织中Ar蛋白表达水平。用放射免疫分析法测定血清睾酮（T）水平。结果B组在mRNA（Shh，Ar）表达水平和血清睾酮水平较对照组有明显的降低（P〈0．05），C组在mRNA（Shh，Bmp4，Ar）和蛋白（Ar）表达水平以及血清睾酮水平都较对照组有明显的降低（P〈0．05）。结论DBP通过对生殖结节（GT）生长发育早期的Shh信号系统，以及晚期的雄激素信号系统发生作用而影响GT的生长发育，诱导尿道下裂的发生。     

尿道下裂仔鼠阴茎蛋白质组学分析及膜联蛋白A3的表达变化研究

目的:探讨邻苯二甲酸二丁酯(DBP)胚胎期暴露对仔鼠阴茎组织蛋白表达谱的影响,分离并鉴定差异表达蛋白质。验证膜联蛋白A3在仔鼠阴茎中的表达变化,初步探讨其在尿道下裂发生中的作用。方法:SPF级SD孕鼠20只,妊娠14～18d(GD14～GD18),随机均分为2组,实验组给予DBP按800mg/kg染毒孕鼠,对照组按大豆油5ml/kg,2组孕鼠均每天灌胃1次连续5d。出生后3d取出实验组尿道下裂雄性仔鼠阴茎和对照组正常雄性仔鼠阴茎,提取总蛋白,进行二维凝胶电泳分离和图像分析,筛选出的差异蛋白质点利用质谱技术进行鉴定,并运用Western印迹和免疫组化法分析膜联蛋白A3在仔鼠阴茎中的表达变化。结果:共筛选出31个差异表达蛋白,其中17个通过质谱分析和SwissProt蛋白数据库检索得到鉴定,包括丙酮酸激酶M2、α烯醇化酶、膜联蛋白A3等。膜联蛋白A3相对表达量在尿道下裂组为1.851±0.014(n=10),正常对照组为2.603±0.012(n=10),两组的差异具有显著性(P<0.05)。膜联蛋白A3主要定位于仔鼠尿道上皮细胞。结论:运用蛋白质组学方法,建立了DBP孕期暴露致尿道下裂雄性仔鼠与正常仔鼠阴茎蛋白质差异表达谱系。膜联蛋白A3的表达变化可能在尿道下裂尿道沟融合障碍过程中发挥重要作用。     

Notch1在邻苯二甲酸二丁酯致大鼠尿道下裂发生中的表达

目的:利用邻苯二甲酸二丁酯(DBP)胚胎期暴露诱导子代SD大鼠发生尿道下裂,检测Notch1在尿道下裂胎鼠与正常胎鼠生殖结节(GT)中的差异表达,探讨其在DBP致大鼠尿道下裂发生中的作用。方法:SD孕鼠20只,随机分为两组,实验组和对照组各10只。妊娠(GD)14～18 d,实验组和对照组分别给予DBP 800 mg/(kg.d)和大豆油2 ml/(kg.d)灌胃,孕第19天剖宫产后记录雄性胎鼠出生体重(BW)、肛门和生殖器之间的距离(AGD)及尿道下裂发生率。取DBP组中发生尿道下裂的雄性胎鼠和对照组中的雄性胎鼠,分为尿道下裂组与对照组,提取生殖结节,用Western印迹法和免疫组化法分析Notch1的表达。结果:实验组子代雄性胎鼠体重为(4.40±0.30)g,对照组为(6.11±0.40)g,实验组较对照组明显下降(P<0.05);实验组AGD为(2.17±0.18)mm,对照组为(3.28±0.16)mm,实验组较对照组明显缩短(P<0.05),尿道下裂发生率为42.9%。Notch1在尿道下裂组的相对表达量为0.671±0.021,正常对照组为1.327±0.031,差异有统计学意义(P<0.05);Notch1主要定位于生殖结节上皮细胞,尿道下裂组染色强度明显弱于对照组。结论:染毒期间DBP对雄性子鼠有明显毒性作用,改变了生殖结节中Notch1的表达,可能影响细胞的增殖和凋亡及上皮-间质转化(EMT),导致尿道下裂的发生。     

邻苯二甲酸二丁酯导致雄性大鼠子鼠尿道下裂畸形及其机制研究

目的:邻苯二甲酸二丁酯(DBP)孕晚期染毒诱导雄性SD大鼠子鼠发生尿道下裂畸形,并进一步探讨DBP致雄性子鼠发生尿道下裂的分子作用机制。方法:孕鼠20只,怀孕14~18 d,随机分为对照组和DBP染毒组,每天分别予大豆油及DBP 750 mg/kg灌胃。出生后(PND)1 d,统计雄性子鼠中尿道下裂的发生率,观察尿道下裂雄性子鼠生殖结节的病理学改变。放射免疫法检测血清雄激素水平,实时荧光定量PCR和Western印迹检测生殖结节中雄激素受体(AR)、音猬因子(Shh)、骨形态发生蛋白4(Bmp4)和成纤维细胞成长因子8(Fgf8)的mRNA和蛋白表达水平。结果:对照组中无尿道下裂发生,DBP染毒组中雄性子鼠尿道下裂发生率为43.6%,病理学检查发现生殖结节典型的尿道下裂畸形。尿道下裂雄性子鼠的血清雄激素水平[(0.49±0.05)ng/ml]较对照组[(1.12±0.05)ng/ml]明显降低(P0.05),生殖结节中AR、Shh、Bmp4和Fgf8基因的mRNA水平(0.50±0.05、0.65±0.07、0.42±0.05、0.46±0.04)较对照组(1.00±0.12、1.00±0.15、1.00±0.13、1.00±0.12)明显降低(P0.05),蛋白水平(0.34±0.05、0.51±0.07、0.43±0.05、0.57±0.04)较对照组(1.00±0.09、1.00±0.12、1.00±0.11、1.00±0.13)明显降低(P0.05)。结论:孕晚期DBP染毒可以导致雄性子鼠发生尿道下裂畸形。DBP通过降低雄性子鼠血清雄激素水平和生殖结节中AR、Shh、Bmp4、Fgf8基因的表达,影响生殖结节的正常发育,是DBP导致尿道下裂畸形发生的可能机制。     

转录激活因子3在糖皮质激素与邻苯二甲酸二丁酯联合作用致大鼠尿道下裂发生中的表达变化

目的 利用糖皮质激素与邻苯二甲酸二丁酯(DBP)联合作用致大鼠尿道下裂,观察转录激活因子3(ATF3)在正常大鼠生殖结节(GT)与不同严重程度尿道下裂大鼠生殖结节中的差异表达,并探讨ATF3在尿道下裂发生中的作用机制.方法 将SD孕鼠加只随机均分为4组,在妊娠14～18 d(GD14～18)分别给予:A组:大豆油灌胃2 ml/d;B组:地塞米松(DXM)0.1 mg/(kg·d)皮下注射;C组:DBP 700 mg/(kg·d)灌胃;D组:DXM 0.1 mg,/(kg·d)皮下注射加DBP 700 mg/(kg·d)灌胃.GD19将各组孕鼠处死剖腹统计雄性仔鼠出生体质量(BW)及肛门生殖器间距离(AGD),取雄性仔鼠生殖结节,运用免疫印迹法和免疫组织化学方法分析ATF3的表达.结果 DXM单独作用于孕鼠仅表现为仔鼠BW下降,但与DBP组合作用却能显著增强DBP的致畸作用,导致雄性仔鼠BW及AGD/BW明显减少.ATF3在A组的蛋白相对表达量为0.351±0.012,B组为0.336±0.015,C组为0.603±0.014,D组为0.851±0.016,C组或D组与A组,以及C组与D组之间的差异均有统计学意义(P＜0.05);免疫组织化学结果显示ATF3主要定位于生殖结节的尿道上皮和侧翼间质,C组和D组免疫组织化学染色强度明显强于A组,而D组染色强于C组.结论 DXM联合DBP进一步增强了DBP的致畸作用,提示尿道下裂的发生是环境中多因素协同作用的结果,ATF3在对照组与实验组生殖结节中的表达有明显变化,且随着尿道下裂程度的加重而表达增高,ATF3的高表达影响了生殖结节的发育及尿生殖褶的融合,这可能是尿道下裂发生的机制之一.

Abstract：

Objective To verify the differential expression of ATF3 in the genital tubercle (GT) of rats with hypospadias induced by maternal exposure to Di-n-butyl phthalate (DBP) or DBP + dexamethasone (DXM) in order to further explore the mechanism of hypospadias. Methods Forty pregnant SD rats were randomly divided into four groups which were given: group A, soybean oil oral gavage 2 ml/day;DBP (700 mg/kg gavage) + DXM (0. 1 mg/kg sc-injection) during gestational day (GD) 14-18. Pregnant rats were killed at GD 19, birth weight (BW) and anogenital distance (AGD) were recorded in the fetal rats, GT was harvested to verify the expression of ATF3 by Western blotting and immunohistochemistry. Results DXM alone had no effect except to reduce BW but could increase the teratogenic effect of DBP when combined with DBO. The BW and AGD/BW in group D were significantly lower than in group C. The expression of ATF3 had statisticall significant difference between groups C or D and group A, as well as between groups C and D (P ＜ 0. 05). ATF3 was mainly located in the urethral epithelium and mesenchyme. Conclusion These results suggests that exposure to common environmental chemicals in combination with environmental endocrine disrupters may increase the risk of male reproductive abnormalities,such as hypospadias. Up-regulation of ATF3 may affect urethral fold fusion, and then lead to the occurrence of hypospadias.     

波形蛋白在邻苯二甲酸二丁酯诱导大鼠尿道下裂中的表达研究

目的 利用邻苯二甲酸二丁酯(DBP)胚胎晚期暴露导致子代SD胎鼠发生尿道下裂的动物模型,检测波形蛋白(Vimentin)在尿道下裂与正常雄性胎鼠阴茎组织中的差异表达,以进一步探讨DBP诱导尿道下裂发生的作用机制.方法 妊娠14～18d SD大鼠20只,随机分为两组,实验组和对照组分别给予DBP和大豆油800mg/(kgod)灌胃,在妊娠19d (GD19),提取雄性胎鼠阴茎组织,用免疫印迹法检测Vimentin 蛋白的表达,用免疫组织化学方法分析其定位及染色情况.结果 Vimentin蛋白在尿道下裂雄性胎鼠中的相对表达量为0.583±0.031 (n =10),在对照组的相对表达量为0.702±0.054 (n =10),差异有统计学意义(P＜0.05).Vimentin蛋白主要定位于尿道上皮细胞,尿道下裂胎鼠染色强度较对照组明显减弱.结论 孕晚期大鼠暴露于DBP后改变了雄性胎鼠尿道组织中Vimentin的表达,影响了尿道上皮细胞的迁移和尿生殖褶的融合,这可能是尿道下裂发生的原因之一.     

有机磷农药-敌敌畏诱导大鼠尿道下裂动物模型睾丸组织的病理学初步观察

目的:探讨环境内分泌干扰物-有机磷农药敌敌畏导致尿道下裂发生的可能机制。方法:30只SD大鼠随机分为两组:实验组(20只)和对照组(10只)。于孕12~17 d分别以敌敌畏10 mg/(kg.d)和生理盐水灌胃。实验组产雄仔鼠88只,其中22只发生尿道下裂,对照组产雄仔鼠33只,无尿道下裂发生。实验组(尿道下裂鼠)和对照组各5只仔鼠喂养至性成熟,取其睾丸组织作常规石蜡切片分别用HE染色和用抗大鼠Calretin in抗体作SP免疫组化染色检测。结果:尿道下裂大鼠睾丸组织中间质细胞(Leyd ig)数量明显减少,生精小管数及形态无明显改变;尿道下裂大鼠睾丸组织中Calretin in阳性的Leyd ig细胞数明显少于对照组。结论:孕SD大鼠染毒敌敌畏农药后能导致雄性仔鼠睾丸组织中的Leyd ig细胞数量减少,推测环境内分泌干扰物质敌敌畏诱导大鼠尿道下裂发生的可能机制是敌敌畏的毒性作用致使胎鼠睾丸中的Leyd ig细胞受损,导致胎鼠睾丸产生的睾酮水平降低,在尿道形成过程中发生障碍而产生尿道下裂。     

尿道下裂大鼠模型的建立及莠去津对大鼠尿道下裂发病率的影响

目的建立SD大鼠尿道下裂动物模型,探讨莠去津对大鼠尿道下裂的发病率有无影响。方法将受孕的SD大鼠120只,随机分为6组,每组20只。于孕期第11～16天时,每天分别给予莠去津25mg/kg、莠去津100mg/kg、莠去津200mg/kg、植物油1ml/kg、非那雄胺10mg/kg、非那雄胺20mg/kg。给药后即刻观察并记录动物的反应情况,于孕期第0、3、6、9、12、15、18、20天称重以调整剂量。于仔鼠出生后第21天,检测其有无尿道下裂,并记录其肛生殖窦距离及其体重。结果200mg/kg莠去津组、10mg/kg非那雄胺组、20mg/kg非那雄胺组仔鼠出生时尿道下裂发生率分别为10.23%、28.30%、67.03%;25mg/kg莠去津组中有2只母鼠的胚胎着床后发育停滞。各组雄性仔鼠的肛生殖窦距离（AGD）与其体重存在正相关关系。结论①大鼠仔鼠尿道下裂表现为阴茎头和（或）阴茎腹侧裂开、尿道口异位,并有肛生殖窦距离的变窄,SD大鼠孕期第11～16天管饲每天10mg/kg及20mg/kg非那雄胺,可以建立尿道下裂动物模型。②莠去津对SD大鼠是一种胚胎致畸物。     

苯甲酸雌二醇诱导小鼠尿道下裂动物模型的建立

目的:建立苯甲酸雌二醇诱导的小鼠尿道下裂模型,为研究雌激素致尿道下裂的分子作用机制提供进一步研究的基础。方法:100只ICR孕小鼠随机分成A、B、C、D、E 5组,每组20只,在孕12~16 d连续5 d各组每只皮下分别注射苯甲酸雌二醇0、0.2、1、5、25 mg.kg-1.d-1,出生时观察仔鼠死亡率。每组取2只母鼠所生的雄性仔鼠,解剖观察仔鼠睾丸位置、前列腺发育情况。出生后4周,观察有无尿道下裂和隐睾。结果:A、B、C、D、E各组仔鼠死亡率分别为21.6%、21.5%、41.4%、56.6%、75.0%。新生鼠解剖,各组均未发现前列腺,C、D、E组睾丸均在下极,A、B组睾丸均在膀胱两侧。仔鼠尿道下裂发生率分别为0、0、3.3%(2/60)、20.0%(8/40)、23.0%(6/26),尿道下裂发生率D、E组与A组间差异有显著性(P<0.05),而D、E组间差异无显著性(P>0.05);隐睾发生率分别为0、0、6.6%(4/60)、30.0%(18/60)、61.5%(16/26),隐睾发生率A、C组间差异无显著性(P>0.05),D、E组与A组间差异有显著性(P<0.05),而D、E组之间差异无显著性(P>0.05)。结论:孕鼠注射大剂量的雌激素可以诱导出仔鼠尿道下裂的模型,C组诱导率较低,D、E组诱导率接近,但是E组母鼠及其仔鼠死亡率均较高,因此5 mg.kg-1.d-1苯甲酸雌二醇用药量为小鼠尿道下裂造模的适宜剂量。     

孕激素过度暴露对尿道下裂发病的实验研究

目的探讨胚胎早期母体孕酮过度暴露与尿道下裂发生的关系。方法随机三组SD大鼠(每组5只),分别于孕12～19d连续皮下注射生理盐水,50mg/kg和100mg/kg黄体酮,雄仔出生14d后分别测量体重肛门生殖器距离(AGD),测各组尿道下裂的发生率。同时,将两实验组中尿道下裂仔鼠睾丸与对照组睾丸采用免疫组织化学APA法进行切片的VEGF染色和VEGF阳性细胞计数。结果(1)对照组产生雄仔鼠17只,无尿道下裂表现;50mg/kg组尿道下裂发生率42.86%(6/14);100mg/kg组的发生率为65.00%(13/20)。三组的AGD分别为(6.36±0.56)mm,(5.33±0.81)mm,(4.65±0.67)mm。尿道下裂发生率与AGD各组间均存在差异(P=0.001);(2)对照组、50mg/kg组、100mg/kg组染色阳性细胞分别为(26±3)、(15±2)、(7±2),组间P<0.05,差异有显著性。结论试验结果支持怀孕早期的母体孕酮过度暴露是尿道下裂发生的危险因素之一。     

半乳糖凝集素-3在邻苯二甲酸二丁酯致大鼠尿道下裂发生中的表达

目的 利用邻苯二甲酸二丁酯(DBP)胚胎期暴露导致SD大鼠尿道下裂发生的动物模型,观察半乳糖凝集素-3(galectin-3)在尿道下裂与正常大鼠生殖结节中的差异表达,探讨其在DBP致大鼠尿道下裂发生中的作用.方法 SD孕鼠20只,随机分为2组,妊娠14～18 d(GD14～18)实验组和对照组分别给予DBP 800 mg/(kg·d)和大豆油2 ml/d灌胃,GD19剖宫产后统计子鼠数、雄性子鼠出生体质量(BW)及肛门生殖器距离(AGD)、尿道下裂发生率,取两组雄性子鼠生殖结节,运用免疫印迹法和免疫组织化学方法分析galectin-3的表达.结果 尿道下裂仅实验组发生,发生率为40%,每窝子鼠数、雄性子鼠BW及AGD/BW在实验组分别为(8.90±1.25)只、(5.12±0.24)g、(0.57±0.03)mm/g,在对照组分别为(13.20±1.46)只、(6.72±0.42)g、(0.66±0.07)mm/g,差异有统计学意义(P＜0.05);galectin-3在尿道下裂组的相对表达量为0.603±0.014(n=10),正常对照组为0.851±0.015(n=10),差异有统计学意义(P＜0.05);galectin-3主要定位于生殖结节上皮细胞,尿道下裂组染色强度明显弱于对照组.结论 染毒期间DBP对雄性子鼠有明显毒性作用,改变了生殖结节中galectin-3的表达,影响了上皮细胞增殖、凋亡及尿生殖褶的融合.     

Morphological features of external genitalia in hypospadiac rat model: 3-dimensional analysis

PURPOSE: We examined 3-dimensionally the process of external genitalia formation in the experimental hypospadiac rat model. MATERIALS AND METHODS: We administered 7.5 mg flutamide daily, a blocker of androgen receptor, into the abdomen of naturally pregnant female Sprague-Dawley rats from gestational days 14 to 20 to produce a hypospadiac rat model. The control group consisted of male offspring not exposed to flutamide. The fetal phallus was obtained at gestational days 17.5, 19.5 and 21.5. We observed them by scanning electron microscopy. RESULTS: In the 17.5-day-old embryo the projection on the ventral side of the phallus was observed from the base of the phallus to the coronary sulcus in the control rat. This finding demonstrated that the urethra develops from the base of the phallus to the coronary sulcus. On the other hand, this projection was not observed in the hypospadiac rat and the urethra was not seen on the ventral side of the phallus. In the 19.5-day-old embryo the ventral preputial closure appeared in the proximal phallus of the control rat but not of the hypospadiac rat. In the control rat the scrotum was discerned in the perineum and divided by a median fold (the raphe). The raphe reached the base of the phallus, where it was concurrent with the preputial fold that covers the urethra. On the other hand, the hypospadiac rat did not have a raphe and the hollow at the base of the phallus appeared to give rise to the future orifice of the urethra. In the 21.5-day-old embryo the control rat prepuce completely surrounded the distal phallus, whereas that of the hypospadiac rat was dorsally hooded with a ventral cleft and the external appearance resembled the morphology of human penoscrotal hypospadias. CONCLUSIONS: This hypospadiac rat model is considered valuable for further studying penile growth and differentiation, and the molecular mechanisms in external genitalia formation in hypospadias.     

Combined exposure to anti-androgens causes markedly increased frequencies of hypospadias in the rat

The incidence of hypospadias is increasing in young boys, but it remains unclear whether human exposure to endocrine disrupting chemicals plays a role. Risk assessment is based on estimation of no-observed-adverse-effect levels for single compounds, although humans are exposed to combinations of several anti-androgenic chemicals. In a mixture (MIX) study with three androgen receptor antagonists, vinclozolin, flutamide and procymidone, rats were gavaged during gestation and lactation with several doses of a MIX of the three chemicals or the chemicals alone. External malformations of the male reproductive organs were assessed on PND 47 using a score from 0 to 3 (normal to marked) for hypospadias. Markedly increased frequencies were observed after exposure to a MIX of the three chemicals compared to administration of the three chemicals alone. Anogenital distance at PND 1, nipple retention at PND 13, and dysgenesis score at PND 16 were highly correlated with the occurrence of hypospadias, and MIX effects were seen at doses where each of the individual chemicals caused no observable effects. Therefore, the results indicate that doses of anti-androgens, which appear to induce no hypospadias when judged on their own, may induce a very high frequency of hypospadias when they interact in concert with other anti-androgens.     

青春期前邻苯二甲酸二丁酯持续暴露对大鼠睾丸发育的影响

目的:研究青春期前邻苯二甲酸二丁酯(DBP)持续暴露对睾丸发育的影响。方法:21日龄断乳青春期前雄性SD大鼠随机分为对照组(n=24)和实验组(n=54),每日分别用玉米油或DBP玉米油溶液灌胃,DBP暴露剂量分别为50mg/(kg.d)(低剂量组,n=18)、200mg/(kg.d)(中剂量组,n=18)和600mg/(kg.d)(高剂量组,n=18),各组动物持续暴露14、21、28d后(即PND35,PND42和PND49)断颈处死。记录大鼠体重变化,检测睾丸重量和体积、附属性器官重量及附睾精子,化学发光免疫分析法检测血清睾酮含量,苏木精-伊红染色观察睾丸组织形态学变化,测量生精小管平均直径及进行睾丸活检评分。结果:低剂量组PND35少量生精小管生精细胞排列紊乱,PND42和PND49睾丸、附属性器官发育及生精功能正常;中剂量组PND35和PND42生精细胞排列紊乱、数目减少,PND49生精小管内可见各级生精细胞及精子,睾丸未见萎缩,附属性器官发育正常;高剂量组大鼠体重增长减缓,血清睾酮水平低下,睾丸生精小管变性萎缩,生精上皮发育阻滞,生精细胞大量凋亡坏死,青春期大鼠睾丸萎缩,无精子,附睾、前列腺和精囊等附属性器官发育迟缓。结论:青春期前DBP持续暴露可损害睾丸组织发育和正常生精功能形成,其毒性效应具有剂量依赖性,高剂量DBP持续暴露引起的睾丸毒性在青春期前发育过程中不可修复,而低中剂量暴露引起的睾丸毒性在PND49之前可完全或部分逆转性恢复。     

The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats

Background/purpose

Androgen has been shown to regulate inguinoscrotal testicular descent. This study aims to clarify the effect of one of the major endocrine disrupters, vinclozolin (V), on both gubernacular migration and inguinoscrotal testicular descent in rats.

Methods

Time-pregnant rats were segregated into 2 groups. In group I, the rats were administered 200 mg/kg/d of V by gavage on days 15 to 18 of gestation. In group II, the rats were administered the same volume of solvent and were used as controls. At birth, the anogenital distance was measured in pups, and gubernacular migration was examined at 10 days of age in some of male offspring. Next, the incidence of testicular descent and the growth of external genitalia were investigated in the remaining male offspring at 60 days of age. The χ² test was used for statistical analysis of the results.

Results

At birth, the anogenital distance (AGD) index decreased significantly more in group I than in group II in male offspring. However, there was no significant difference in the AGD index between the 2 groups in the female offspring. At 10 days of age, an aberrant migration of the gubernaculum was found in the 51.5% of V-treated rats in group I. At 60 days of age, the incidence of cryptorchidism was 57.7% in group I and 0% in group II (P < .05). In addition, hypospadias with cleft phallus and pseudo vagina with a blind pouch also were observed in some of the V-treated rats.

Conclusions

Prenatal administration with V thus caused intrauterine defects, which resulted in testicular maldescent caused by the induction of an aberrant migration of the gubernaculum associated with an abnormal extension of the processus vaginalis, and this may have been caused by the antiandrogenic effect of V in utero.     

Spermatogenesis, fertility and sexual behavior in a hypospadiac mouse model

PURPOSE: Administering of flutamide to pregnant mice causes hypospadias in male offspring. We investigated spermatogenesis, fertility and sexual behavior in this hypospadiac mouse model. MATERIALS AND METHODS: Male offspring exposed to flutamide during the embryonic period were divided into hypospadiac group 1 and normal external genitalia group 2. Control group 3 consisted of male offspring not exposed to flutamide. We analyzed the spermatogenesis, epididymides sperm motility, in vitro fertilization rate and sexual behavior of each mouse. RESULTS: There were no significant differences in the weight of the testes or mean seminiferous tubular diameter in the groups. The number of apoptotic germ cells per unit area was not significantly different in the 3 groups. In groups 1 to 3 there were no significant differences in the mean epididymides sperm motility rate plus or minus standard deviation (62.6% +/- 10.0%, 57.2% +/- 7.0% and 67.0 +/- 7.6%) or in the in vitro fertilization rate (52%, 48% and 48%, respectively). However, there were significant differences in groups 1 to 3 in mean mounting frequency (0, 29 +/- 4.0 and 12.4 +/- 4.5 times per hour) and mean intromission frequency (0, 24.4 +/- 3.5 and 3.8 +/- 1.5 times per hour, respectively). Females coupled with group 1 or 2 male mice did not achieve pregnancy. CONCLUSIONS: These results suggest that spermatogenesis, sperm motility and fertilization in vitro were unaffected in hypospadiac mice but sexual motivation and arousal were deficient.     

Scrotal base distance: A new key genital measurement in males with hypospadias and cryptorchidism

Background:Anogenital distance (AGD) in both humans and animals is a known reflection of fetal endocrine effect on genital virilization and the related abnormalities, including cryptorchidism and hypospadias. However, we introduce here and investigate scrotal base distance (SBD) as a sensitive genital anthropometric biomarker in human infants with cryptorchidism and hypospadias, which are considered early manifestations of testicular dysgenesis syndrome. We aim to assess SBD in patients with cryptorchidism or hypospadias against healthy subjects.Material and methods:Patients with hypospadias (n = 61, age 17.4 ± 6.3 months) or cryptorchidism (n = 51, age 11.4 ± 4.8 months) were enrolled for assessment of SBD, AGD, and penile length; and compared with a cohort of 102 full-term healthy boys for standard ritual circumcision by measuring age-specific standard deviation scores.Results:Patients having hypospadias had lower mean SBD, AGD, and penile length standard deviation scores than the control group (p < 0.01). These values in patients with cryptorchidism were longer than mean values in boys with hypospadias (p < 0.01) and shorter than mean values in the control group.Conclusions:We showed that SBD, AGD, and penile length were lower in patients with cryptorchidism or hypospadias compared to normative data measured from a control group of healthy boys for ritual circumcision. These results enforce the use of SBD as an objective anthropometric measurement and a viable biomarker to assess the effects of fetal endocrine imbalance on male external genitalia development.