Effects of zinc deficiency on morphogenesis of the fetal rat eye

Maternal zinc deficiency during pregnancy results in a high frequency of fetal eye malformations in the Long-Evans rat. In this study we examine the development of the eye from days 12 through 21 of gestation in conceptuses of dams fed deficient or adequate levels of zinc and also examine maternal plasma and conceptus zinc concentrations during this period. Dams were fed diets containing 0.5 (0.5 Zn group), 4.5 (4.5 Zn group), or 100 (100 Zn AL group) micrograms zinc per gram diet ad libitum, or 100 micrograms zinc g-1 diet in amounts restricted on a daily basis to the intake of matched animals from the 0.5 Zn group (100 Zn RI group). Conceptuses were removed and maternal plasma was collected on days 12, 14, 16, 19 and 21 of gestation. Maternal plasma and conceptus zinc concentrations reflected maternal dietary zinc level, with dam plasma Zn concentrations in the order of 0.5 Zn group less than 4.5 Zn group less than 100 Zn group on all days. A similar pattern held for embryo/fetus zinc, except for days 19 and 21, at which times the 0.5 Zn and 4.5 Zn fetuses had similar zinc concentrations. Histological examination of the developing eye of 0.5 Zn fetuses on days 12 and 14 revealed that invagination of the optic cup was often deficient, and that closure of the choroid fissure did not occur, resulting in colobomata and retinal folding visible at term. A very few fetuses were found at term to be anophthalmic or have only remnants of ocular tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of glycoprotein synthesis inhibitor on myelination in rat cerebellum

Effects of a glycoprotein synthesis inhibitor on myelination were investigated in rat cerebellum. The glycoprotein synthesis inhibitor, tunicamycin (TM), was injected intracranially into newborn rats. The activity of 2,3-cyclic nucleotide 3-phosphodiesterase (CNPase) in the cerebellum was significantly reduced in 2-week-old animals and was restored to the normal level by age 3 weeks. When TM was injected into newborn rats every 3–4 days for a total of 6 times, CNPase activity was still low at 3 and 4 weeks. Immunohistochemical stainings for CNPase and myelin-associated glycoprotein (MAG) were performed on paraffin sections of multiple-TM-injected cerebellum at 3 weeks. The intensity of the staining with MAG antiserum in the white matter was clearly decreased in TM-treated cerebellum compared with the control. The myelin in the granule cell layer was poorly stained with CNPase antiserum in TM-treated cerebellum. Subcellular fractionation was carried out and the CNPase activity in each fraction was measured. The CNPase activity in the myelin fraction (P₂A) from the TM-treated cerebellum was significantly lower than that in the control. In contrast, the activity in the synaptosomal (P₂B) and microsomal (P₃) fractions from the multiple-TM-injected cerebellum was higher than in those from the controls. Polyacrylamide gel electrophoretic patterns of the P₂A fractions were analyzed. The P₂A fraction from TM-treated cerebellum contained less Wolfgram protein than the control. These results suggest that glycoprotein synthesis plays certain roles in myelination in the central nervous system.     

Effects of 60 Hz electric and magnetic fields on the development of the rat cerebellum

The effects of extremely low frequency (ELF) electromagnetic (EM) fields on the maturation of the rat cerebellum were studied. Newborn rats were exposed to 60 Hz electric and magnetic fields under three different combinations in a specially constructed apparatus. The pups were irradiated for 7–8 h daily, with a 30-min interruption for nursing. Pups were kept with their mothers for the remainder of the time. After approximately 1, 2, or 3 weeks of exposure, the pups were killed. Control pups were sham exposed. The somatic growth of the irradiated rats did not show any significant difference from shamexposed controls. At 1 kV/m and 10 gauss exposure, there was a small but statistically significant decrease in cerebellar mass. In rats exposed at 1 kV/m and 10 gauss, DNA and RNA levels were significantly higher than those in shara-exposed controls at 6 and 13 days of age, but at 20 days, these two biochemical constituents were similar in both groups of rats. The ELF-EM treatment had no effect on protein and cerebroside concentrations. In terms of age effects. DNA and RNA exhibited increases from 6 to 13 days of age, and declined from 13 to 20 days. Protein and cerebroside levels exhibited increases during the 6–20 day periods. In rats exposed at 100 kV/m and 1 gauss, the DNA levels were initially less than those of sham-exposed controls at 8 days of age, reached approximately the same levels at 14 days, and then were higher than those of controls at 22 days. There was. therefore, a significant ELF-EM effect as well as a significant interaction between age and ELF-EM exposure. In terms of age effects, DNA levels for both control and exposed animals increased from 8 to 14 days. From 14 to 22 days, DNA levels of exposed rats continued to increase while those of the controls decreased. This age effect was significant. RNA levels in both groups of animals showed increases from 8 to 14 days of age, but the increase was less for the irradiated animals than for the controls. From days 14 to 22. RNA levels for both groups showed a reduction, but the decrease was greater in the irradiated than in control rats. ELF-EM treatment significantly reduced protein levels at 8 days of age. but at 14 to 22 days, protein levels of exposed rats were higher than those of controls. The cerebroside levels were not affected by exposure treatments but increased with the age of the animals. Exposure to 100 kV/m and 10 gauss did not exert any effect on the concentrations of DNA, RNA, protein, and cerebroside at all three time points examined. Both DNA and RNA exhibited increases with age from 6 to 13 days, and leveled off from 13 to 20 days. Protein and cerebroside levels also showed corresponding increases with the age of the animals. Morphological observations revealed no detectable changes in the irradiated animals in any experimental group. Thus, only biochemical studies indicate that exposure at certain ELF-EM field combinations induces alterations in cerebellar maturation. These changes were clearly detectable in the early postnatal period but gradually diminished with time. ©1993 Wiley-Liss, Inc.     

Effect of alkylating drugs on rat cerebellum

Ultrastructural and histochemical studies were performed on Wistar rats treated during 6 days with Cyclophosphamide 50 mg/kg/day or Lomustine 20 mg/kg/day. Changes in cerebellum after both drugs were similar. Most severe ultrastructural lesions were found in glia, mainly astroglia cells localized in perivascular area. Cytoplasm of these cells was on the large area devoid of organelles. Neurons were less affected but in Purkinje cells mild ultrastructural lesions were found. Increased activity of glucose-6-phosphatase and decreased of succinate dehydrogenase activity in these cells suggest that cytostatic treatment disturbed metabolism in the neurons.     

The localization of mercury and metallothionein in the cerebellum of rats experimentally exposed to methylmercury

Summary Rats were dosed with methylmercuric chloride, either by gastric gavage (5 × 10 mg kg^-1 body weight over a 15-day period), or in their drinking water (20 mg methylmercuric chloride l^–1 for 14 or 42 days). Localization of mercury within the cerebellum was performed with a silver physical development technique, and metallothionein with dinitrophenyl hapten-sandwich immunohistochemistry. Mercury was detected in structurally undamaged Purkinje neurones and adjacent Bergmann glial cells; no mercury was detected in granule cells even though these small cells nearest the Purkinje layer had a high incidence of pyknotic nuclei. In general, metallothionein was detected mainly in Bergmann glial cells, Purkinje cells, astrocytes and glial cells of white matter; no metallothionein was detected in granule cells. We hypothesized that the resistance of Purkinje cells to methylmercuric chloride reflects their ability to transform organic mercurials to inorganic mercury that, in turn, induces the synthesis of radical-scavenging metallothionein molecules.     

Effects of selective doses of x-irradiation on the levels of several amino acids in the cerebellum of the rat

The cerebella of rats were exposed to selective doses of low levels of x-irradiation beginning on day 4, 8, or 12 following birth. The doses of x-irradiation given on days 12, 13, and 15 (12–15X group) resulted in a 24% reduction in the wet weight of the cerebella; the doses given on days 8, 9, 11, 13, and 15 (8–15X group) resulted in a 57% weight reduction; the doses given on days 4, 5, 7, 9, 11, 13, and 15 (4–15X group) resulted in a 67% weight reduction. The schedule of x-irradiation begun on day 12, which prevented the acquisition of the late-forming granule cells, reduced the levels (nmole/mg dry tissue weight) of alanine (22%) and glutamate (10%), and increased the levels of glycine (15%), GABA (13%), and taurine (71%), with respect to control values. The schedule begun on day 8, which prevented the acquisition of stellate and granule cells, reduced the levels of alanine (15%), glutamate (12%), and taurine (21%), and increased the levels of glycine (102%) and GABA (56%). The schedule begun on day 4, which prevented the acquisition of basket, stellate, and granule cells, reduced the level of glutamate (15%) and increased the levels of glycine (186%) and GABA (78%). The levels of alanine and taurine in the cerebella of the 4–15X group were the same as control values. The level of aspartate in the cerebella of the 3 groups of x-irradiated animals was not significantly different from control values. The consistent reduction in the level of glutamate as a function of the number of doses of x-irradiation is suggestive that glutamate may have a higher level in the granule cells than in other cells in the cerebellum, and that the higher level may be a reflection of a possible excitatory transmitter role for glutamate. In addition, the data are interpreted in terms of taurine being associated with the stellate cells and possibly serving as a transmitter for these inhibitory interneurons.     

The morphogenesis of myeloschisis in the rat

Pregnant Sprague-Dawley rats were utilized in this study. They were separated into two groups. In the control group, a single intragastric dose of distilled water was given on the 11th day of gestation. In the test group, a single intragastric dose of ethylenethiourea (ETU), 240 mg/kg was given on the same day of gestation. Embryos were recovered 12, 24, 36, and 48 hours after ETU and distilled water administration, and were prepared for scanning electron microscopy and light microscopy. The posterior neuropore of rat fetuses in the control group closed completely on gestation day 12.5. However, the closure of posterior neuropore in ETU-induced fetuses is shown to have been disrupted 12 hours after ETU administration. Marked neural tissue overgrowth in the posterior neuropore resulted in neural fold eversion and finally produced a picture of lumbosacral myeloschisis on day 13 of gestation. Our observation implies that myeloschisis is induced by non-closure of the neural fold, not by reopening after its proper closure.     

Effect of nonionizing radiation on the purkinje cells of the rat cerebellum

In one experiment, Sprague Dawley rats (16–21 days of gestation) and their offspring were exposed to 100-MHz (CW) electromagnetic radiation at 46 mW/cm² (SAR 2.77 mW/g) for 4 h/day for 97 days. In another experiment, the pregnant rats were irradiated daily from 17 to 21 days of gestation with 2450-MHz (CW) microwaves at 10 mW/cm² (SAR 2 mW/g) for 21 h/day. In a third experiment, 6-day-old rat pups were irradiated 7 h/day for five days with 2450-MHz radiation at 10 mW/cm². Equal numbers of animals were sham irradiated in each group. Quantitative studies of Purkinje cells showed a significant and irreversible decrease in rats irradiated during fetal or fetal and early postnatal life. In animals exposed postnatally, and euthanized immediately after irradiation, significant decrease in the relative number of Purkinje cells was apparent. However, restoration apparently occurred after forty days of recovery.     

氨基酸对白念珠菌形态学影响的研究

目的初步探讨单个氨基酸对白念珠菌形态学的影响。方法用0．67％的酵母氮源基础培养基和2％葡萄糖配制成SD合成培养基,37％恒温摇床培养,研究单个天然氨基酸对白念珠菌形态学的影响,并分别通过不添加碳源和厌氧条件下培养观察对精氨酸诱导的菌丝的影响。结果在含10mmol／L的L－精氨酸的SD液体培养基中,可见大量的菌丝。在含10mmol／L的L一半胱氨酸、L．苏氨酸、L－缬氨酸和L－色氨酸的sD液体培养基中,可见典型的酵母细胞,未见菌丝。在含10mmol／L的其他单个氨基酸的SD液体培养基中可见混合的酵母和菌丝结构。在不含氨基酸或含各种天然氨基酸的SD固体培养基上,白念珠菌的菌落均光滑。但在含10mmol／L的L-精氨酸固体培养基上,光滑的菌落周围可见小的突起,镜下可见菌丝。无氧条件下,无论有无碳源,含精氨酸的SD培养液中白念珠菌只能形成酵母细胞,生长部分受到抑制。结论精氨酸可以诱导白念珠菌菌丝形成,厌氧条件下精氨酸不能诱导白念珠菌菌丝形成。     

Neurotoxicological effects of deltamethrin on the postnatal development of cerebellum of rat

Deitamethrin (DLT) has been accepted to be 10,000 times less toxic to man than to insects. While toxicity of DLT in adult animals has been studied using biochemical and electrophysiological tools, reports on its developmental neurotoxicity are rather scanty. Wistar rat pups were exposed to DLT (0·7 mg/kg body wt/day, i.p., dissolved in propylene glycol from postnatal day 9–13. Equal number of age matched pups were used as vehicle controls. The animals were weighed and perfused intracardially on postnatal days 12,15,21, and 30 and their brains dissected out. Cerebellum along with the brainstem was separated by a transverse section at the tectal level and processed for morphometric and toxicological studies. The micro-and inter-neurons in the cerebellum are known to differentiate and mature, both morphologically and biochemically, during the postnatal life of rats. Postnatal exposure to DLT has been observed to delay the cytogenesis and morphogenesis of these neurons. In addition to this, damage to the developing vasculature has also been recorded in the form of thrombus and haemorrhage. Focal degeneration and spongy appearance of the tissue in the vicinity of the damaged blood vessels have also been recorded. The study has opened up several questions on the safety of this substance to the pregnant mothers and infants in the habitats where this substance is in use for vector control.