Pollution and developmental abnormalities of Atlantic fishes

Synopsis Planktonic eggs of Atlantic mackerel,Scomber scombrus, provide evidence that pollution is associated with mortality, malformation, and abnormal chromosome division of fish embryos developing about the surface of the U.S. Atlantic coast. Embryo data are substantiated by the finding that adults of mackerel, windowpane flounder,Scophthalmus acquosus, and winter flounder,Pseudopleuronectes americanus, from more polluted coastal areas also have higher frequencies of mitotic abnormality than those from less polluted regions of the Mid- and North Atlantic. No ontogenetic interval escapes contamination. All are likely to be adversely influenced, resulting in considerable direct and indirect cumulative effect on total early-life survival. Development of genetic and epigenetic resistance to reproductively harmful influences of contaminants may interfere with other modifications in structure and function necessitated by natural environmental fluctuations, changes in climate, and by fishing itself.     

Thyroid stimulating hormone enhancement of bovine oocyte maturation in vitro.

Efforts to improve bovine embryonic development in vitro involved study of effects of thyroid stimulating hormone (TSH) alone or in combination with LH on bovine oocyte maturation (IVM). Putative effects were assessed by observing cumulus expansion (CE), fertilization (IVF), and development to morulae/blastocysts (M/B). Effects of prolactin (PRL) were also investigated. Variables for the 24-hr IVM interval were no hormone (control), TSH (0.1, 0.5, or 1.0 micrograms/ml) or PRL (10, 100, or 1000 micrograms/ml), luteinizing hormone (LH) (0, 10, or 100 micrograms/ml) + TSH (0.1 or 0.5 micrograms/ml), and serum (20%, v/v) + 0.5 micrograms TSH/ml; data were from 4-5 trials for each IVM treatment. Higher proportions of oocytes exhibited complete CE with hormones or serum than without (P less than 0.05). All oocytes (with and without CE) were inseminated with heparin-capacitated sperm. A higher proportion of inseminated oocytes cleaved after IVM with 0.5 micrograms TSH/ml (53.4%) than for other TSH treatments (P less than 0.05). The combination of TSH (0.1 and 0.5 micrograms/ml) with 10 micrograms LH/ml for IVM enabled higher proportions (P less than 0.05) of ova to fertilize (67.4 and 69.2%) than did medium alone (28.3%), LH (10 micrograms/ml) alone (54.1%) or serum + 0.5 micrograms TSH/ml (55.6%). No improvement in proportions undergoing fertilization was seen after addition of TSH to 100 micrograms LH/ml for IVM. Frequency of CE and cleavage did not differ among PRL treatments. More M/B developed from cleaved ova after IVM with LH or TSH than with PRL or no hormone (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of cryoprotectant and genetic selection for body fat content on embryonic cryosurvival in mice

Lines of mice selected for high (HF) or low (LF) 12-week epididymal fat pad weight as a percentage of body weight were used to investigate the effects of genotype, two cryoprotectants [glycerol (GLY) and propylene glycol (PG)] and genotype x cryoprotectant interaction on cryosurvival of four and eight-cell embryos. Embryos were collected from selection lines and reciprocal crosses of selection lines (HFLF and LFHF) and frozen by established slow-cool methods. Embryos were thawed for 40s at room temperature and then placed in a 37° C waterbath for 1 min. Cryoprotectant was diluted from embryos with either 0.5 M sucrose (GLY-treated) or 1.0 M sucrose (PG-treated). Post-thaw survival was measured as the percentage of embryos developing to 36 h (PTS36), 48 h (PTS48) and hatched blastocyst (PTSHB), respectively. Non-frozen controls were cultured concurrently with frozen embryos. No significant genotype or genotype x cryoprotectant interaction effects were found. Results of the embryo freezing study indicated that selection for high or low fat content did not affect the ability of embryos to survive cryopreservation. There was no indication of embryo heterosis for post-thaw survial. Embryos frozen with GLY survived the freeze-thaw stress significantly better than those frozen in PG (P < 0.05). In vitro development of non-frozen controls at 36 and 48 h did not vary significantly among lines, but in vitro development was significantly different among lines at the hatched blastocyst stage (P < 0.05). Linear contrasts showed that the embryonic genome was responsible for differential in vitro development at the hatched blastocyst stage between these selected lines (HF > LF; P < 0.05); asymmetric response also occurred in that both HF and LF exceeded the unselected control line (P < 0.05).The research reported in this publication was funded by the North Carolina Agricultural Research Service (NCARS), Raleigh, NC 27695-7643. Use of trade names in this publication does not imply endorsement by the NCARS of the products named, nor criticism of similar ones not mentioned Present address: Department of Animal Science, University of Tennessee, Knoxville, TN 37901, USA     

Effects of cryopreservation on the survival rate of the seven-band grouper (Epinephelus septemfasciatus) embryos

The effects of cryopreservation and the vitrification solution on the embryo hatchability of the seven-band grouper Epinephelus septemfasciatus were evaluated in this study. Six small molecule cryoprotectants (PG, MeOH, Gly, DMF, DMSO and EG) and four macromolecular cryoprotectants (glucose, fructose, sucrose and trehalose) were used to determine the embryo toxicity levels. Results showed that the embryo survival rate was higher when the PM (24% PG + 16% MeOH):Gly ratios were 3:1 and 4:1. Further experiments showed that the embryo survival rates in PMG3S (35% PMG3 + 5% sucrose) and PMG3T (35% PMG3 + 5% trehalose) were relatively higher, which are 29.24 ± 10.81% and 27.01 ± 3.39%, respectively. When treated with PMG3S and PMG3T by using 5-step method, embryos at somite stage and tail-bud stage shrank in the first 6 min and gradually recovered in volume to the original. This indicated the successful permeation of the vitrification solutions into cells. Then, embryos at the embryoid body formation stage, the somite stage and the tail-bud stage were cryopreserved with PMG3S and PMG3T. In total, 82 floating embryos were obtained, 14 of which developed further, with 8 embryos at the tail-bud stage developing to the heartbeat stage, 4 embryos at the body formation stage development to the somite stage, and 2 embryos at the somite stage hatched to larval fish.     

Environmental modulation of calcium and phosphorus metabolism in embryonic snapping turtles (Chelydra serpentina)

Summary Eggs of common snapping turtles (Chelydra serpentina) were incubated on wet (–150 kPa water potential) and dry (–950 kPa) substrates in a laboratory study assessing the effects of the hydric environment on patterns of mobilization of calcium and phosphorus by developing embryos. We found that embryos developing in wet environments withdrew nutrients from their yolk faster, grew more rapidly, and incubated longer than embryos exposed to dry environments. Turtles developing in both environments absorbed calcium from the yolk at similar rates and depleted the yolk of almost its entire reserve of calcium prior to hatching. Calcium withdrawn from the yolk was supplemented with calcium mobilized from the eggshell, but embryos in wet environments obtained substantially more calcium from the eggshell than did those in dry settings. Embryos obtained all of the phosphorus used in skeletogenesis from the yolk, but those incubating in wet environments mobilized phosphorus from this compartment more rapidly than did those in dry settings. Exposing embryonic snapping turtles to wet environments apparently allows them to make more efficient use of the transitory source of calcium in the eggshell than is possible in dry environments. However, the residual yolk in hatchlings from both wet and dry environments contains too little calcium to support the growth of hard and soft tissues in neonates at rates similar to those characterizing the growth phase of development in embryogenesis.     

斑马鱼胚胎和肿瘤细胞评价细胞毒药物活性比较研究*

目的:比较斑马鱼胚胎和肿瘤细胞作为药物筛选模型的优缺点.方法:采用MTT法检测顺铂、紫杉醇、阿霉素、5-氟尿嘧啶四种药物对HL-60和Hela细胞的增殖影响;同时,观察药物对斑马鱼胚胎发育的影响.结果:阿霉素、顺铂及紫杉醇作用于HL-60及Hela细胞的IC50均显著高于作用于斑马鱼胚胎的LD50;而5-FU作用于肿瘤细胞和斑马鱼胚胎的结果与其它药物相反;四种抗肿瘤药物对斑马鱼胚胎的生长发育均有致畸作用.结论:斑马鱼胚胎作为细胞毒类药物筛选模型,对于抗微管类药物较为敏感,但对于抗代谢药敏感性较肿瘤细胞差.     

In vitro plant regeneration from quality protein maize (QPM)

Breeding efforts to obtain more nutritious maize materials aimed at alleviating dietary deficiencies in developing countries have resulted in an improved maize germplasm known as quality protein maize (QPM). Quality protein maize has higher contents of tryptophan, lysine, and leucine than common maize, but suffers from some major agronomic drawbacks found in common inbred maize lines, such as susceptibility to insect pests and fungal and bacterial diseases and herbicide sensitivity. The development of a reproducible and efficient protocol for tissue culture of QPM is expected to solve some of these deficiencies. In this work, we have evaluated different formulations for in vitro induction of morphogenic responses in three QPM lines developed by the International Maize and Wheat Improvement Center (CIMMYT): CML (CIMMYT maize line)-145, CML-176, and CML-186. Only CML-176 and CML-186 have proven to be responsive to the in vitro conditions considered in this work, with CML-176 showing the highest efficiency in regenerable callus formation and growth. N6C1 medium was found to be efficient for in vitro culture of QPM, whereas no plants could be regenerated by using MPC medium. From CML-176 embyogenic calli cultured on N6C1 medium, we were able to regenerate up to 0.3 plants per 500 mg fresh weight (FW) callus. Further modifications in this experimental protocol, including the replacement of 3,6-dichloro-o-anisic acid with 2,4-dichlorophenoxyacetic acid and modification of the N6C1 vitamin balance, significantly increased the regeneration response of the induced calli, with up to 16.8 and 9.3 plants recovered per 500 mg FW callus for CML-176 and CML-186, respectively.     

BTB蛋白结构与功能研究进展

BTB(Broad-complex, Tramtrack, and Bric-a-brac)蛋白家族存在于痘病毒以及几乎所有真核生物中,该类蛋白为多结构域蛋白,最为显著的特征是含有高度保守且能介导蛋白与蛋白间相互作用的BTB结构域。BTB蛋白具有多种功能,其功能特异性取决于BTB蛋白中其他结构域以及它的互作蛋白。BTB蛋白广泛参与转录调节,染色质重组装,细胞骨架调控和泛素化降解等过程,与胚胎发育,器官形成,信号转导以及免疫调节等生理过程密切相关。除此之外,多种疾病如癌症,神经系统和骨骼肌系统疾病等的病理过程也与BTB蛋白相关。本文以蛋白结构为基础总结了该家族的共性规律并重点论述了BTB蛋白在转录调节以及泛素化降解过程中发挥的重要作用,以期为后续研究提供重要参考。     

Vitrification of early-stage bovine and equine embryos

The objectives of this study were to: (1) determine an optimal method and stage of development for vitrification of bovine zygotes or early embryos; and (2) use the optimal procedure for bovine embryos to establish equine pregnancies after vitrification and warming of early embryos. Initially, bovine embryos produced by in-vitro fertilization (IVF) were frozen and vitrified in 0.25 mL straws with minimal success. A subsequent experiment was done using two vitrification methods and super open pulled straws (OPS) with 1- or 8-cell bovine embryos. In Method 1 (EG-O), embryos were exposed to 1.5 M ethylene glycol (EG) for 5 min, 7 M ethylene glycol and 0.6 M galactose for 30 s, loaded in an OPS, and plunged into liquid nitrogen. In Method 2 (EG-DMSO), embryos were exposed to 1.1 M ethylene glycol and 1.1 M dimethyl sulfoxide (DMSO) for 3 min, 2.5 M ethylene glycol, 2.5 M DMSO and 0.5 M galactose for 30 s, and loaded and plunged as for EG-O. Cryoprotectants were removed after warming in three steps. One- and eight-cell bovine embryos were cultured for 7 and 4.5 d, respectively, after warming, and control embryos were cultured without vitrification. Cleavage rates of 1-cell embryos were similar (P > 0.05) for vitrified and control embryos, although the blastocyst rates for EG-O and control embryos were similar and higher (P < 0.05) than for EG-DMSO. The blastocyst rate of 8-cell embryos was higher (P < 0.05) for EG-O than EG-DMSO. Therefore, EG-O was used to cryopreserve equine embryos. Equine oocytes were obtained from preovulatory follicles. After ICSI, injected oocytes were cultured for 1-3 d. Two- to eight-cell embryos were vitrified, warmed and transferred into recipient's oviducts. The pregnancy rate on Day 20 was 62% (5/8) for equine embryos after vitrification and warming. In summary, a successful method was established for vitrification of early-stage bovine embryos, and this method was used to establish equine pregnancies after vitrification and warming of 2- to 8-cell embryos produced by ICSI.