Morphology of oocyte and follicle destruction by polycyclic aromatic hydrocarbons in mice.
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Biomedical subjects
Publications and source records attributed to D R Mattison.
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The polycyclic aromatic hydrocarbon (PAH) benzo(a)pyrene (BP) reduced the fertility of DBA/2N mice in a dose-dependent fashion. Control mice produced offspring at a rate of 0.91 pups/mouse per week. Treatment with BP at doses of 10, 100, 200, and 500 mg/kg decreased offspring production rates to 0.61, 0.20, zero and zero pups/mouse per week, respectively. BP also destroyed primordial oocytes in similarly treated mice. Treatment with BP at doses of 10, 50, 100, and 500 mg/kg destroyed 20%, 58%, 88%, and 100%, respectively, of the primordial oocytes in DBA/2N mouse ovaries. Dose-response curves for fertility reduction and primordial oocyte destruction were identical. The threshold for fertility reduction was 3.4 mg/kg and for primordial oocyte destruction it was 2.7 mg/kg. The 50% effect doses for fertility reduction and primordial oocyte destruction were 25.5 mg/kg and 24.5 mg/kg, respectively. Although BP reduced fertility and destroyed primordial oocytes, it had no effect on the ovarian weight response to pregnant mare's serum gonadotropin (PMSG), consistent with the observations that BP at these doses does not destroy growing or preovulatory oocytes or follicles. The similarity of the dose-response curves for fertility reduction and primordial oocyte destruction suggested that their mechanism of action is similar and resides within the ovary. As BP had no effect on ovarian response to PMSG, the effect was likely to reside outside the regulatory mechanisms controlling ovulation. These data suggest that the site of fertility reduction by BP may reside in the mechanisms of fertilization, implantation, or early conceptus development.
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Destruction of mouse oocytes in primordial and small primary follicles in response to treatment with 3-methylcholanthrene (MC) was studied at the ultrastructural level. Four-week old C57Bl/6N (B6) strain mice received a single injection of 80 mg/Kg MC in corn oil intraperitoneally. Controls received only corn oil. Ovaries from animals were prepared for light and electron microscopic examination at specified intervals after treatment. The number of primordial follicles remained constant in control animals. In contrast, their number decreased significantly (P less than 0.01) by three days, and they were depleted by seven days after MC treatment. Subtle degenerative modifications were noted in the ooplasm of primordial follicles two days after treatment. These changes consisted of vesiculation of mitochondrial cristae, increased electron density of the mitochondrial matrix, myelin structures in lipid droplets and in mitochondria. More advanced stages of degeneration of primordial follicles were characterized by further vesiculation or disappearance of mitochondrial cristae, chromatin clumping, and increased density of the ooplasm. Small primary follicles had undergone similar initial degeneration as primordial follicles. In more advanced stages of degeneration nuclear and cytoplasmic contents condensed, endoplasmic reticulum, Golgi complex and mitochondria swelled, small vesicles and multivesicular bodies appeared. In the most advanced stages of degeneration of small primary follicles it appeared that small portions of the oocyte were engulfed by the surrounding follicular cells. It is concluded that exposure of B6 mice to a single dose of MC results in atresia of oocytes in primordial and small primary follicles. Ultrastructurally, these degenerating oocytes of treated mice looked much like the spontaneously atretic oocytes in untreated animals.
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The rodent ovary contains an enzyme system(s) capable of metabolising poly-cyclic aromatic hydrocarbons to reactive electrophilic intermediates known to cause cytotoxicity, mutation, and cancer. If the human ovary contains similar enzyme systems, metabolic activation of environmental chemicals could explain the earlier menopause in cigarette smokers and the higher incidence of ovarian cancer in industrialised areas.
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Oxytocin (OX) has been suggested as a signal for parturition. Although OX is produced by both mother and fetus, concentrations are higher in umbilical than maternal blood. In addition, umbilical artery OX concentrations (15-40 pg/ml) are higher than umbilical vein (4-12 pg/ml) and maternal (1-10 pg/ml) concentrations. The umbilical A-V difference suggests that placental uptake and transport may be one path from fetal (F) to maternal (M) circulation. However, this difference may also reflect placental oxytocinase activity, which is known to metabolize biologically active peptides such as OX. We have investigated placental transport of OX from F to M and M to F circulation using in vitro dually perfused isolated cotyledons from term human placenta. Term human placentae from uncomplicated pregnancies were obtained immediately after delivery. A single peripheral cotyledon and corresponding lobule was cannulated and perfused. After stabilization and demonstration of adequate M to F perfusion-perfusion overlap, we studied the transport of OX (3H) with 14C-inulin (14C-IN) as permeability reference in both M to F (n = 8) and F to M (n = 6) directions during 2 h of perfusion. In addition to the higher tissue uptake observed in M to F than F to M transport direction as measured by the drop in the concentration of both 3H-OX and 14C-IN in the circuits in which both compounds were added, the same trend was found for the transfer rates of both compounds. These transfer rates which reflect the permeability of placental tissue to OX and IN were 15.17 +/- 2.79 (mean +/- SD) and 6.28 +/- 0.93 microliters/min/g (M to F) and 11.79 +/- 1.77 and 4.91 +/- 0.81 microliters/min/g (F to M). Although the permeability of both compounds is higher in the M to F than in the F to M transport direction, comparing these permeability values with respect to their molecular weight (MW) showed a significant correlation when known permeability values of polar compounds between MW 60 and 68,000 daltons were included. This correlation indicates that OX crosses the placenta in both directions by simple diffusion. High-performance liquid chromatography analysis showed that there is little evidence of placental metabolism and degradation of OX over the period of these experiments. Oxytocin is the main therapeutic drug that is frequently used in obstetrics for the induction of labor and parturition. Under such circumstances and with respect to the placental permeability results, oxytocin could reach the fetal circulation.