3d-reconstruction Of Newborn Mice Genitourinary System And The Effect Of Des On Fetal Mice Testicular Morphology

Objective:In the recently, with the high incident of congenital disorders of the genital system, more and more research which study on the causes and mechanism of the diseases is advanced. However, it is difficult to make a particular standard of the quantitative morphology of congenital abnormality of genital system in the previous studies, especially with complicated genital system and the systemic and multiple abnormalities. Even though can't be used in the research on the effects of environmental low-dose chemicals. More accurate techniques, methods and standards of research in this way are urgent needed. Three-dimensional computer reconstruction as an emerging technology can not only reconstruct the organs by serial slices, but also calculate the length, volume, surface area of the organs. The reconstructed mouse genitourinary system was build by 3D-reconstruction software with serial slices and treated with the synthetic estrogen diethylstilbestrol (DES) in this study. And then the morphology, position and structure of the testes were observed or measured. To explore the exact standard quantitative morphology of the genital system included the abnormalities.Methods:Thirty 8-10 weeks old Kunming female mice were divided into three groups randomly: normal group (not given any drug), experimental groupⅠ(DES was given 1μg/kg/d) and experimental groupⅡ(DES was given 100μg/kg/d). Pregnant mice were injected subcutaneously from GD9 (gestation day) to GD17. The newborn mice, when they were delivered were collected at once for the determination of sexuality. And among six male mice were randomly selected from each group. All the selected newborn mice were treated as follows:the low parts of abdomen were fixed in situ and set to transected serial slice. All the slices (HE prepared and 40μm thick) ware recorded serially under microscopy as JPG form. The 3D-reconstruction was made by the software of Photoshop and 3D-doctor. The volume and surface area of the testis was get and being statistically analyzed thereafter.Results:1. In the serial transects of normal newborn mice, the tissue images proved that the kidney and testes of normal newborn mice were all in round shape and their curves were smooth. The positions of right kidney and testes were all higher than the left one. The testes located in two sides of the bladder neck, and far away from the kidney. The testis and epididymides narrowed downwards and gradually varied and coalesced into gubernaculum. In the experimental groups, the testes volumes of newborn mice were smaller than those of normal groups. They later near the kidney and correspondence to DES dosage.2. The 3D reconstruction picture of genitourinary system and morphological parameters were acquired:①In the normal group, the surface area of left and right testes was about 2621923.89±280270.28 and 2651522.14±53701.92μm2 respectively; the volume of left and right testes was about 432910679.57±50133711.68 and 418385403.85±20078887.04μm3 respectively,②In the experimental group I, the surface area of left and right testes was about 2728634.03±200344.17-and 2728634.03±200344:17μm2 respectively; the volume of left and right testes was about 442275396.27±51415726.79 and 432652932.27±33081719.62μm3 respectively.③In the experimental group II, the surface area of left and right testes was about 2148691.89±280270.28 and 2083414.40±118167.77μm2 respectively; the volume of left and right testes was about 269457600.00±48909033.77 and 289652459.20±36903360.51μm3 respectively. The surface area and volume of testes in the experimental groupⅡwere all smaller obviously than that of normal and experimental group I (P<0.05).Conclusions:1.3D-reconstruction of the genitourinary system in newborn male mice with serial slices is feasible.2.1t not only can reconstruct the 3D picture of the genitourinary system in newborn male mice but also acquire precise morphological parameters to distinguish the fine difference of the tiny organ morphology.3. Exposed to high does DES make the newborn mice testes smaller and later near the kidney corresponded to DES dosage.