Embryolethality in rats caused by retinoic acid.

Retinoic acid (25 mg/kg) administered by gavage to rats at 216 hours of gestation killed almost all conceptuses by 288 hours and all by full term. The embryolethal dose50 was 12.3 mg/kg. Embryos died from damage to the allantois leading to agenesis or hypogenesis of the chorioallantoic placenta. Suppression of cell division in, disturbed fluid entry into, and impaired normal vascularization of the allantois underlay the abnormality, the predominant element depending principally on when exposure occurred. Hydremia (Br. J. Exp. Pathol., 57:525-541, '76) affected over 50% of the sacs. Relating data from the literature to resorption patterns suggested that 25 mg/kg of retinoic acid raised a lethotoxic level (approximately 2 micrograms/ml) of retinoate in the plasma for about 5 hours. This, together with asynchronous development, was used to help explain why groups of embryos responded to the teratogen for 18 hours longer than single embryos and why exposure 18 hours before the allantois on average appears, killed some young. Comparison showed that retinoic acid was 4.8 times as embryolethal as retinol. Otherwise, each behaved qualitatively similarly, suggesting that either retinol acts through retinoic acid or via a common path entered independently by each. Placental agenesis is well documented in Soviet literature. It is almost unknown in Western teratology even though it is probably the most important cause of embryonal death following teratogenic procedures around the time of placentation.

The chronology of somites in rat embryos.

To establish their chronology in rats the somites of 902 embryos were counted. They were the products of 89 pregnancies terminated at three hourly intervals from 231 hours to 288 hours and then six hourly until 324 hours. Somites form at a virtually constant rate of one somite per 1.87 hours. This is incorporated in the regression equation (formula; see text) where y = number of somites, x = hours of gestation from 06.00 on the morning after overnight mating. The greatest variation in the maturity of embryos within any time-determined sample was nine somites (= 16.8 hours) and the least was three somites (= 5.6 hours). Some uses for the data in experimental teratology are suggested.

A model for ulnar dysmelia.

The treatment of pregnant rats with the carbonic anhydrase inhibitor, acetazolamide, produced gross limb malformations primarily affecting the forepaw, but also producing variable ulnar dysmelia. Analysis of the cytoarchitecture of the ulnar dysmelic limbs showed the presence of cartilaginous and fibrocartilaginous connections between the ulnar and radial chondroepiphyses, with variable deformation of the radial chondroepiphysis by the tethering effect (although the growth plate, per se, did not appear affected at the stage of development studied). The extremely variable experimental appearances duplicated most of the variations seen in the human disease analogue, and suggest this drug-induced embryopathy may be useful as a model for the study of postaxial forelimb deformities in the postnatal phase in order to adequately assess the structural changes of disparate growth between radius and ulna due to the presence of the cellular continuity between the two distal chondroepiphyses.

Placental agenesis, embryonal hydraemia, embryolethality and acute hypervitaminosis A in rats.

Acute maternal hypervitaminosis A established on Day 9 of gestation in Sprague-Dawley-derived rats caused a dose-related increase in the resorption of implants. The median embryolethal dose was 189,000 i.u./kg. In addition to suppression of the allantois leading to placental agenesis, damaged embryos showed retarded somatic development and hydraemia, all apparent 24 h after treatment. At about Day 11 the hydraemia involved the visceral wall of the yolk sac causing death of the embryo soon after. The fluid in the vitelline vessels continued to collect until Day 13 when it absorbed following necrosis of the wall of the yolk sac. Two mechanisms are suggested for the embryonal hydraemia: either the excess fluid resulted from a permeability disorder induced by the vitamin A; or it was retained metabolic water or water specifically absorbed to inflate the allantois and, being unused for this purpose, it pooled in the blood vessels of the embryo. The yolk sac hydraemia is more likely to have followed injury to the proximal endoderm.