吗啡对鸡胚胎发育的影响

目的：研究吗啡对胎动、心率、孵化率、孵化时间、雏鸡体重等的影响。方法：以气室给药的方式给鸡胚注射吗啡,记录胎动、心率、孵化率、孵化时间、雏鸡体重。结果：吗啡可以缩短雏鸡的孵化时间,降低雏鸡的孵化率,并导致雏鸡出现运动障碍;20mg／kg吗啡剂量和12—16胚龄的给药时间,鸡胚孵化率最高,残疾率最低;吗啡导致胚胎心率加快,胎动减少（P〈0．05）。结论：吗啡对胚胎发育有损伤作用,损伤程度与吗啡剂量和给药时间有关。     

Proliferating ability,morphological development and acetylcholinesterase activity of the neural tube cells in early chick embryos

Summary Using ³H-thymidine autoradiography and AChE histochemistry at the electron microscopic level on the same sections, the interrelationships between loss of proliferating ability, morphological development and increase of AChE activity during the course of differentiation of the neural tube cells were investigated in early chick embryos. The neural tube wall consisted of spindle-shaped cells with no AChE activity, weakly positive spindle-shaped cells showing AChE activity in the cisternae of the nuclear envelope and in a few short profiles of r-ER, moderately positive spindle-shaped cells showing AChE activity in the nuclear envelope and in a moderate number of r-ER profiles and intensely positive large round cells showing AChE activity in the nuclear envelope and in a large number of r-ER profiles. Nuclei of the AChE-negative, weakly positive and moderately positive cells were located in the ependymal layer (matrix). The AChE-intensely positive cells were in the mantle layer. The AChE-negative and weakly positive cells were capable of proliferation and were regarded as undifferentiated neuroepithelial cells. In contrast, the moderately positive and intensely positive cells were no longer capable of proliferation and were considered to be neurons. These findings indicate that the r-ER increases rapidly in amount and volume in newly formed neurons soon after their final cell division, and that AChE increases in the neurons in parallel to the development of the r-ER.     

Effects of 100-Hz magnetic fields with various waveforms on the development of chick embryos

Summary Chick embroys were exposed during their 52 first hours of development to 100-Hz magnetic fields. Sinusoidal, square and pulsed waveforms were used at average field strengths from 0.1 A/m to 80 A/m. After exposure, the embryos were examined for abnormalities and classified by the developmental stages. When bipolar oscillations (oscillating at both sides of the zero-level) were used, the percentage of abnormal embryos was significantly increased above 1 A/m. In exposure to unipolar square waves, no significant effect on the percentage of abnormalities could be demonstrated. The developmental stage was possibly affected by unipolar square waves at 0.1 A/m, all other field strengths and wave-forms being apparently ineffective.     

In silico development of new acetylcholinesterase inhibitors

In this work, we made use of fragment-based drug design (FBDD) and de novo design to obtain more powerful acetylcholinesterase (AChE) inhibitors. AChE is associated with Alzheimer’s disease (AD). It was found that the cholinergic pathways in the cerebral cortex are compromised in AD and the accompanying cholinergic deficiency contributes to the cognitive deterioration of AD patients. In the FBDD approach, fragments are docked into the active site of the protein. As fragments are molecular groups with a low number of atoms, it is possible to study their interaction with localized amino acids. Once the interactions are measured, the fragments are organized by affinity and then linked together to form new molecules with a high degree of interaction with the active site. In the other approach, we used the de novo design technique starting from reference drugs used in the AD treatment. These drugs were broken into fragments (seeds). In the growing strategy, fragments were added to each seed, growing new molecules. In the linking strategy, two or more separated seeds were linked with different fragments. Both strategies combined produced a library of more than 2 million compounds. This library was filtered using absorption, distribution, metabolism, and excretion properties. The resulting library with around six thousand compounds was filtered again. In this case, structures with Tanimoto coefficients >.85 were discarded. The final library with 1500 compounds was submitted to docking studies. As a result, 10 compounds with better interaction energy than the reference drugs were obtained.     

Effects of RNA inhibitors on the development of Drosophila embryos permeabilized by a new technique

The molecular analysis of Drosophila embryogenesis has been hindered by the impermeable nature of the vitelline membrane, which has made it difficult to introduce exogenous substances into the developing embryo. We have developed a modification of the permeabilization technique of Limbourg and Zalokar ('73) in which octane is used to permeabilize the vitelline membrane and dimethylsulfoxide is used to facilitate the transport of exogenous substance across the cell membranes. The procedure is highly effective (ca. 95%) and is consistent with a high frequency of normal development. We have used this technique to analyze the effect in vivo of four inhibitors of RNA synthesis (alpha-amanitin, actinomycin-D, rifampin, and rifamycin SV) on the embryogenesis of Drosophila. We have found that there are characteristic stage-specific alterations in the sensitivity of the embryo to these inhibitors which is reflected both by changes in the ID50 dosages and by changes in the developmental abnormalities caused by the drugs. Embryos aged 2-12 hours old undergo a developmental arrest within 30 minutes after application of the inhibitors. Embryos older than 12 hours are able to develop for 140+ minutes after treatment before arresting. The effects of these drugs are consistent with the idea that there exists a definite sequential program of gene activity that is necessary to the normal embryonic development of Drosophila.     

Effects of the acute and chronic administration of chlordiazepoxide on the development of spontaneous motility in chick embryos

The effects of the acute and chronic administration of chlordiazepoxide on spontaneous motility and on the reactivity of the generator of embryonic motility were studied in chick embryos from the 4th to the 19th day of incubation. 1. The acute administration of chlordiazepoxide (5 mg/kg e.w.) significantly depressed spontaneous motility from the 13th day of incubation. 2. The chronic administration of chlordiazepoxide (12.2 mg/kg e.w./24 h) from the 4th to the 8th, 12th and 16th day of incubation enhanced the reduction of the spontaneous motility of 17-day-old embryos. 3. The chronic administration of chlordiazepoxide significantly modified the activity of both activators (strychnine, metrazol, bicuculline, picrotoxin) and inhibitors (GABA, chlordiazepoxide) of the spontaneous motility of chick embryos.     

Mechanism of asymmetric ovarian development in chick embryos

In most animals, the gonads develop symmetrically, but most birds develop only a left ovary. A possible role for estrogen in this asymmetric ovarian development has been proposed in the chick, but the mechanism underlying this process is largely unknown. Here, we identify the molecular mechanism responsible for this ovarian asymmetry. Asymmetric PITX2 expression in the left presumptive gonad leads to the asymmetric expression of the retinoic-acid (RA)-synthesizing enzyme, RALDH2, in the right presumptive gonad. Subsequently, RA suppresses expression of the nuclear receptors Ad4BP/SF-1 and estrogen receptor alpha in the right ovarian primordium. Ad4BP/SF-1 expressed in the left ovarian primordium asymmetrically upregulates cyclin D1 to stimulate cell proliferation. These data suggest that early asymmetric expression of PITX2 leads to asymmetric ovarian development through up- or downregulation of RALDH2, Ad4BP/SF-1, estrogen receptor alpha and cyclin D1.     

Proteomic profiling of facial development in chick embryos

Craniofacial disorders are associated with one-third of human birth defects but the underlying molecular and cellular causes remain poorly understood. Proteomics seems well-placed to benefit this medically important area but the scarcity of embryonic tissues poses a major challenge. In this study, we applied a microsample proteomics strategy to investigate the first branchial arch, an embryonic structure crucial for facial development, and found that proteome analysis is both practicable and informative despite the scarcity of tissue. Exploiting the embryonic chick as a tractable source of accurately staged tissue, we developed a sequential extraction procedure to interface with one-dimensional polyacrylamide gel electrophoresis (1-D PAGE) and 2-D PAGE. In 2-D gels, about 8% of the visible proteome changed between embryonic days 3 and 5, and the identities determined for 21 proteins accorded with the rapid growth during this period. These results led to the first molecular identification of chicken alpha-fetoprotein, and an unusual localisation of vimentin to endoderm. With over 470 protein spots accessible, this comparative proteomics approach has good prospects for providing new markers, functional hypotheses and genes to target in functional tests. A broader value of extending these approaches to facial development in other species and to other areas in embryology can be anticipated.     

Localization of embryonal acetylcholinesterase during the early development of the chick embryo

The presence of "embryonic" acetylcholinesterase activity, as described by Drews (1975) was investigated during early chick embryonic development, mainly in the following systems: a) primitive streak and Hensen's node during gastrulation movements; b) area opaca during blood islets and vessels differentiation; c) mesoderma of lateral laminae, during delamination movements. The demonstration of enzymic activity was performed with slightly modified histochemical methods. The enzyme was thus localized around the nuclei, in the cytoplasm and associated to plasma membrane of cells engaged in morphogenetic movements. The enzyme activity localized at the plasma membrane was supposed to be involved in the regulation of membrane functions concerning intercellular communications, such as inductive message, perhaps mediated by ion fluxes.     

Effects of proteinase inhibitors on preimplantation embryos in the rat

Proteinase inhibitors of microbial origin were injected into the uterine horns of mated rats at 14:00 h on Day 5 of pregnancy (spermatozoa in vaginal smear = Day 1), and 5 or 6 h later the embryos were flushed from the horns and examined. Chymostatin and alpha-MAPI, inhibitors of chymotrypsin-like serine proteinase and thiol proteinases, as well as thiolstatin, an inhibitor of thiol proteinases, significantly inhibited embryo growth. The inhibitory activity of alpha-MAPI on embryonic growth was distinctly greater than that of thiolstatin, although the ID50 values of the two inhibitors to papain are similar. Antipain and leupeptin which are inhibitors of trypsin-like and thiol proteinases, and talopeptin, an inhibitor of metal proteinases, significantly interrupted the removal of the zona pellucida from expanding blastocysts. These results suggest that (1) a chymotrypsin-like proteinase seems to be important to the growth of the embryo, (2) a thiol proteinase may participate in embryonic growth, and (3) a trypsin-like proteinase and a metal proteinase are likely to participate in zonalysis.     

Effects of corticosterone on brain cholinergic enzymes in chick embryos

The effects of corticosterone on the cholinergic enzymes, choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) were studied in the chick embryonic brain. Chick embryos received either 0.25, 0.5, or 1.0 g of corticosterone via the air sac daily for three days during either embryonic days 6 through 8 (E6-E8), of cerebral neurogenesis, or days 10 through 12 (E10-E12), a period of cerebellar neurogenesis. Enzyme activities were determined in cerebral hemispheres, optic lobes, cerebellum and remaining brain at 10, 15, and 20 days of incubation. In embryos treated from E6 to E8, ChAT activity was generally higher at day 10 in cerebral hemispheres and optic lobes (cerebellum was not determined) while AChE activity was not affected. At day 20 ChAT activity of treated chick embryos was lower in the cerebral hemispheres and optic lobes, but not in the cerebellum; AChE activity was higher in the cerebral hemispheres, lower in the optic lobes, and not changed in the cerebellum as compared to controls. However, in embryos treated from E10 to E12 both cerebellar ChAT and AChE activities were higher at day 15 in comparison to controls. These data show that the hormonal effects were most prominent only in the brain areas undergoing neurogenesis during the period of hormonal treatment. Since AChE activity is also present in nonneuronal cells, the observed alterations caused by corticosterone may reflect glial cell responses to the hormone. Whether the hormone affects the final number and/or maturation of cholinergic neurons and/or glial cells remain to be investigated.     

Application and effects of some fat-soluble hormones on the development of chick embryos

Summary An attempt has been made to employ fat-soluble anabolic steroids (testosterone propionate and ethylestrenol) in the manner of water soluble compounds. The substances are injected into the egg proper (i.e., the yolk ball) as close to the embryo as possible. The effects are assessed from the points of view of mortality, malformation and increase in dry and fresh weights. The mortality pattern varies with the substance injected. No malformations were detected. Both testosterone propionate and ethylestrenol cause increase in dry weights at particular doses. Sesame oil which is used as carrier of these substances is itself responsible for some mortality. The experiments also suggest that in lower dosages ethylestrenol somehow counteracts the lethal effect of sesame oil.