Cellular necrosis in zinc-deficient rat embryos.

Abnormal cellular necrosis was studied in 9.5-11.5-day embryos obtained from zinc-deficient rats. At periods of low maternal zinc status induced by a high intake of a zinc-deficient diet, cell death was observed in those regions of the embryo that were most sensitive to teratogenic insult at that time. As the maternal serum zinc level increased during the fasting phase of the feeding cycle, the degree of necrosis decreased, leaving the tissue histologically more normal even though the embryos were grossly malformed. The mitotic index of cells in the neural epithelium and limb buds of zinc-deficient, non-necrotic embryos was found to be elevated, but there was no evidence of blockage at any particular stage of mitosis. It can be hypothesised that during the early stages of organogenesis, periods of low maternal zinc status initiate unscheduled cell death by some as yet undefined mechanism that in turn, gives rise to the morphological anomalies observed later.

A selective stain for mitotic figures, particularly in the developing brain.

A selective stain for mitotic figures is valuable where autoradiographic counting is not required, especially in the developing brain. Most work in this field has been based on conventional nuclear stains which do not differentiate mitotic figures from resting cells by color. Hematoxylin, Feulgen, gallocyanin and Nissl methods have been used particularly. The method described uses a modified Bouin fixative, followed by hydrolysis in 1 N HCl. Mitotic figures are selectively stained using crystal violet, with nuclear fast red as the counterstain for resting cells. The method has been tested using material from postnatal and fetal sheep, guinea pig and rat. Using paraffin mounted serial sections it is applicable to all organs. The method was very successful on developing rat brain, particularly for detail and quantitative estimation in the early stages of prenatal development, which was of primary interest. Nucleated cells of the erythrocytic series, keratin and what appear to be mast cells were found to stain. When nuclear counting or cell recognition were required these did not cause any difficulty, except in prenatal liver. The highly selective method presented stains mitotic figures, in all tissue tested, an intense blue against a background of red resting cells.

Postnatal accumulation of zinc by the rat hippocampus.

Timm's staining material has been detected in the rat hippocampus as early as day 1 postnatally. However, staining was diffuse and widespread and light granulation was observed only in the mossy fiber layer. By day 6 postnatally most diffuse staining had disappeared and the characteristic pattern of granulation had intensified in the mossy fiber layer. Pronounced staining of the mossy fibers became apparent from day 6.Electron microscopic autoradiography indicated that(65)Zn injected intraperitoneally into suckling pups became localized largely in the axons and axon terminals of the mossy fiber layer in the CA3 and CA4 regions of the Horn of Ammon.In vitro studies with hippocampal slices have demonstrated that zinc is accumulated by an active transport system, but the kinetic characteristics of this uptake do not appear to alter with age. Zinc located intracellularly in the hippocampus appears to be associated mainly with large molecular weight ligands, with more than 75% of newly acquired zinc being bound to substances having molecular weights greater than 70,000 Daltons.