| Objective The aim of this research was to investigate the effect of prenatal exposure to diethylstilbestrol (DBS) and the role of ACTIN and PCNA on gubernaculum testis development in Kunming fetal male mice. In this study, histological analysis and scanning electron microscopy (SEM) were used to observe the effect of DES on gubernaculum testis development and testis descent, transmission electron microscopy (TEM) was performed to observe ultrastructural changes in gubernaculum cells, immunohistochemical studies were performed to detect expression and alteration of ACTIN and PCNA during gubernaculum testis development.Material and methods Mature Kunming female mice were divided into six groups randomly, that is, normal group (not given any drug), control group (given DMSO), DES200|ng/kg.d group, DESlOOug/kg.d group, DES50ug/kg.d group and DES25ug/kg.d group (DES dissolved in DMSO). Female mice were placed in individual cages with male mice and the morning when a vaginal plug appeared was designated as gestation day 0 (GDO) of pregnancy. Pregnant mice were injected subcutaneously on GD9 though GDI7 with DES in DMSO or with DMSO alone (controls). Pregnant mice were sacrificed on 6p.m. ofGDI 7 by cervical dislocation. Cesarean sections were performed and fetuses were quickly removed, the embryos were decapitated and the low parts of the embryos were fixed in different solutions according to different methods of this research. The specimens of male offspring were sliced serially in coronal plane, expression of ACTIN and PCNA in gubernaculum testis was detected by immunohistochemical methods, histological changes were observed with light microscope (LM). Morphological and ultrastructural changes of gubernaculum testis were observed with SEM and TEM.Results l.In DES-treated groups, some pregnant mice had a vaginal bleeding before they were sacrificed and had a higher death rate of embryos than normal and control groups. The higher the dose of DES was, the higher death rate of embryos was. 2.Gubernaculum testis of normal and control groups was well developed. Gubernaculum testis consisted of two parts, that is, gubernaculum bulb and gubernaculum cord, gubernaculum bulb was well developed, with an inner mesenchymal core and muscular outerlayer. In contrast, in DES-treated male fetus, gubernaculum testis seemed to be poor developed. The volume of gubernaculum bulb was smaller than that of normal and control groups, and gubernaculum testis appeared to be undifferentiated. 3.ACTIN expression in gubernaculum testis was lower in DES-treated groups than in normal and control groups. The higher the dose of DES was, the lowerACTIN expression was. 4.PCNA expression in guberaaculum testis was lower in DES-treated groups than in normal and control groups. 5. When observed in SEM, guberaaculum testis consisted of gubernaculum bulb and gubernaculum cord. In normal male fetus, guberaaculum bulb was well developed, seemed to be a cylinder, its volume was bigger than that of DES-treated male fetus. Gubernaculum cord was thick and short. The testes were well developed, located next to the bladder, at the bottom of the abdomen. It showed that the transabdominal descent of the testis was taking place in normal male fetus. In DES-treated male fetus, gubernaculum bulb was poor developed, seemed to be a taper, its volume was smaller than that of normal male fetus. Guberaaculum cord was thin and long. The testes were poor developed, located at the lower pole of the kidneys. When observed in TEM, gubernaculum cells of normal group had some bigger myofibrils, the myofibrils were well arranged, fs could be seen in myofibrils, there were abundant organelles in cytoplasm. It showed that metabolism of the cells was active. In contrast, guberaaculum cells of DES-treated group had some smaller myofibrils, the myofibrils were disordered, and few fs could be seen in myofibrils, there were only some sparse organelles in cytoplasm. It seemed that metabolism of the cells was inactive.C... |
PDF Full Text Request