Regional differences in mesenchymal cell morphology and glycosaminoglycans in early neural-fold stage rat embryos.

Rat embryos with two to four pairs of somites (day 9 of gestation) were examined by scanning electron microscopy and Alcian blue staining. The neural folds, which represent only future brain region at this stage, form a pair of elongated hemispheres with a deep neural groove between them. In transverse section the neural ectoderm is biconvex; the cranial mesenchyme cells beneath them are widely separated by extracellular matrix (ECM) and are joined to each other and to the ectodermal basement membrane by fine cytoplasmic processes and strands of ECM material. In contrast, mesenchyme cells close to the primitive streak are closely packed, having broad areas of surface contact and only small amounts of ECM. The nature and distribution of ECM, cell surface, and basement membrane glycosaminoglycans (GAG) were investigated by staining with Alcian blue at specific pH values in combination with enzyme pretreatments, and at various critical electrolyte concentrations. The results indicate that the GAG of the ectodermal basement membrane, mesenchymal ECM and mesenchymal cell surfaces are in continuity with each other and consist largely of hyaluronate and chondroitin sulphates. Differences in morphology and histochemistry of neural fold and primitive streak regions are discussed in relation to their possible morphogenetic significance.

Effect of oxygen concentration on morphogenesis of cranial neural folds and neural crest in cultured rat embryos.

Rat embryos, 9 1/2 days old, cultured with a 5% or 10% O2 gas phase underwent normal or near-normal cranial neurulation; however, culture at 20% or 40% O2 resulted in abnormal morphogenesis of the cranial neural folds from the 9-somite stage onwards, and the brain tube frequently failed to close. Normal morphogenesis was characterized by a narrowing V-shaped profile, development of a slightly concave neuroepithelial surface, and formation of a sharp mediad curvature of the most lateral region prior to midline apposition and fusion. These morphogenetic events were related to cellular changes within the neuroepithelium, namely cell death, onset of neural crest cell migration, and loss of apical microfilament bundles from the most lateral cells. In 20% and 40% O2-cultured embryos, failure of curvature of the neuroepithelium was associated with failure or retardation of the related cellular changes; it may therefore have been due to the maintenance of an excessive rigidity which opposed the forces involved in bringing about the final stage of brain-tube formation. Mitochondria in normal (low O2 and in vivo) embryos were of the anaerobic type, having few cristae; in high O2-cultured embryos they were of the characteristic aerobic type, indicating an adaptation to the abnormal environment.

Changes in the surface features of choroid plexus of the rat following the administration of acetazolamide and other drugs which affect CSF secretion.

The surface of the choroid plexus of the rat was examined by scanning electron microscopy before and after administration of acetazolamide and other drugs (cardiac glycosides and pilocarpine) which affect the rate of secretion of the CSF. In control animals, bleb-covered cells were more common on the IVth ventricle choroid plexus than on the lateral ventricle choroid plexus (20 per specimen compared to 0-3 per specimen). Following administration of acetazolamide the number of bleb-covered cells was reduced significantly (P less than 0-001). The effects of the other drugs were less well defined. Since acetazolamide is known to interfere with the active transport of the extra chloride ions normally added to the CSF in the IVth ventricle, it is suggested that the bleb-covered cells of the IVth ventricle choroid plexus are the specific site for chloride secretion.

Comparison of the effects of retinol and retinoic acid on postimplantation rat embryos in vitro.

Rat embryos were explanted on day 8 or 9 of pregnancy and cultured for up to 48 hours in serum containing added retinol (vitamin A), retinoic acid (vitamin A acid), or absolute ethanol. They were examined morphologically and their protein content determined. Retinoic acid was more teratogenic and growth-retarding than retinol. Electron microscopy of embryos cultured for 30 minutes or one hour revealed that both forms of vitamin A brought about similar ultrastructural effects on the embryonic cells; however, the abnormally large intracellular lipid droplets observed in a previous study following exposure to retinol in vitro and retinyl palmitate in vivo were not observed in embryos exposed to retinoic acid. It is possible that the differential teratogenicity may be due to the inability of the embryonic cells to convert and store retinoic acid in a less teratogenic form.