Testis Histology The method for testis histology is a modification of the protocol by Klinefelter et al. and Parks et al.. The testes from males on PnD 1, 24, and 82 were removed and immersion fixed in 5% glutaraldehyde (#01909; Polysciences, Warrington, PA) in 0.05 M collidine buffer with 0.1 M sucrose overnight at 4°C. […]
Archive for the ‘Dimethanesulfonate’ Category - Part 2
Each morning (between 0700 and 0900 h; lights on at 0600 h) of the 10-day study, the vagina of each of the two females was examined for the presence of a postcoital plug. In the event a plug was observed, the female was designated as GD 0, promptly removed from the box with the male, […]
Ex Vivo Testis Parenchyma Incubations Production of T by testis parenchyma following gestational exposure to EDS was determined by modification of the protocol used by Klinefelter et al. and Parks. Briefly, the right testis of each mouse was removed. The tunica was perforated (PND 24) or removed (PND 82), and the whole testis (PND 24) […]
The sample was decanted into a microcentrifuge tube and stored at 4°C. An aliquot (0.1 ml) was diluted with 0.9 ml of buffer, and sperm were counted using a he-mocytometer slide. Minimally, three hemocytometer chambers were counted per sample.
For immunohistochemical localization of P450 side chain cleavage enzyme in the LCs, adult testes (PND 53) were removed and immediately immersed in Bouin fixative (50 X the volume of the tissue). A 26-gauge needle was used to puncture the tunica albuginea (two nicks on each side of the testis) to facilitate penetration of fixative into […]
Postnatal Necropsy of Male Offspring On PND 1, one male pup per litter was killed via CO2 followed by decapitation. The internal morphology of the urogenital tract and the positioning of the testes were noted. The right testis was removed and placed in a 5-ml glass scintillation vial containing 5% glutaraldehyde in 0.05 M collidine […]
Fetal whole-body T production in male mice was evaluated from GD 15, when morphological identification of the male gonad could first be achieved, through GD 18 just prior to birth. Dosing was terminated the day prior to cesarean section. Thus, a dam killed on GD 16 was dosed from GD 11 to GD 15. Fetuses […]
Treatment Administration Timed pregnant mice were randomly assigned to one of two treatment groups: vehicle (dimethyl sulfoxide [DMSO]: deionized water; 3:7, v/v) or EDS (gift from Sterling Research Group, Rensselaer, NY) (160 mg/kg body weight [BW] per day, in a 30% DMSO solution, i.p.) from GD 11 through GD 17; GD 0 = day of […]
Animals Animal procedures in this study were approved by the Institute Animal Care and Use Committee at the National Health and Environmental Effects Research Laboratory (NHEERL) of the United States Environmental Protection Agency. Timed pregnant CD-1 mice were mated at Charles River Laboratories, Inc. (Raleigh, NC) and delivered to the NHEERL animal facility on Gestation […]
Additionally, adverse functional effects on cultured Sertoli cells, including inhibition of the synthesis of the glycoprotein, transferrin, have been demonstrated at relevant sublethal in vitro doses of EDS (300-500 ^M). Declining levels of this soluble iron transport protein can result in decreased numbers of germ cells and suppression of spermatogenesis.