Mesoderm induction and development of the embryonic axis in amniotes.

The mechanisms controlling the formation of the embryonic axis, and specifically those that give rise to the mesoderm, have received renewed attention recently. In the frog, some of these mechanisms have begun to be elucidated, and several factors have been found to cause uncommitted ectoderm cells to differentiate into mesoderm. All of the factors identified to date are related either to fibroblast growth factor (FGF) or to transforming growth factor beta (TGF-beta). Do the mechanisms that generate the embryonic axis of amphibians also operate in chick and mouse embryos? Here I address how amphibian and amniote embryos might provide complementary pieces of a puzzle.     

Effects of estradiol and progesterone on early embryonic development in aging rats

Regularly cyclic, middle-aged female rats exhibit a decreased incidence of fertility, and those females that are fertile produce small litters. These decreases in fertility and litter size are associated with reduced numbers of normal blastocysts formed and implanted, suggesting that pre- and/or peri-implantation failures may be the causes for these aging-related reproductive declines. The present study examined the relationships and influence of circulating estradiol (E2) and progesterone (P) levels on early embryonic development and implantation in middle-aged rats. Serial blood samples obtained from cannulated, middle-aged pregnant rats revealed minor decreases in plasma P and increases in E2 levels during Days 2-4 of pregnancy, compared to young pregnant rats, resulting in significantly (p less than 0.001) decreased plasma P/E2 ratios. These alterations in endogenous hormone secretion in middle-aged pregnant rats were associated with fewer normal blastocysts on Day 5 of pregnancy and reduced numbers of normally implanting embryos. Correlation analysis further revealed a significant (p less than 0.05) inverse relationship between mean circulating E2 levels and numbers of normal conceptuses on Day 12 of gestation. Moreover, s.c. administration of P implants (in Silastic) to middle-aged pregnant rats increased serum P levels by about 34-40 ng/ml, and significantly (p less than 0.05) reduced the incidence of abnormal embryos before implantation. In contrast, treatment with E2 minipumps produced a sustained rise in serum E2 (by about 7-15 pg/ml) and resulted in the complete absence of embryos in the reproductive tracts by Day 5 of pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inductive interactions in early embryonic development.

This is an update of a previous review (Current Opinion in Cell Biology 2:969-974) in which we discussed recent work attempting to understand the sequence of inductive interactions responsible for establishing the body plan of the early embryo. As before, we concentrate on inductive interactions in amphibian embryos, where significant progress has been made in the past two years. In this update, however, we also consider recent embryological data obtained with amniote embryos such as the chick, together with complementary data provided by genetic analyses of mouse and Drosophila development.     

Epithelial scatter factor and development of the chick embryonic axis

Scatter factor, a recently characterised protein secreted by certain embryonic fibroblasts, affects cultured epithelial by increasing cell motility, the breakdown of cell junctions and cell scattering. The process of gastrulation in higher vertebrate embryos, during which the primitive streak forms, involves an epithelial-to-mesenchymal transformation resembling the effects of the factor on cultured cells. The factor was applied locally to chick embryos, using both scatter-factor-secreting cell lines and inert carriers. We found that scatter factor can generate local supernumerary axial structures resembling primitive streak and/or neural plate and conclude that it may have primitive-streak and/or neural-inducing activity in chick embryos.     

Effects of male accessory sex gland secretions on early embryonic development in the golden hamster

The ventral prostates, dorsolateral prostates, coagulating glands, seminal vesicles and/or ampullary glands were bilaterally removed from adult male hamsters. Removal of these glands did not affect the fertilization rate and cleavage of the embryos at 48 h post coitum (p.c.). Air-dried preparations of the embryos showed a delay in cleavage at 72 h p.c. and a significant number of degenerated embryos was also found in females mated with males from which all the male accessory sex glands had been removed. A significant implantation loss was also observed at 122 h p.c. The results suggest that, in the golden hamster, removal of the male accessory sex gland causes a slower cleavage rate in embryonic development and a significant embryonic loss during pregnancy.     

Effects of PKC activation on the meiotic maturation, fertilization and early embryonic development of mouse oocytes

Protein kinase C (PKC) is a family of Ser/Thr protein kinase widely distributed in eukaryotes. There is evidence that PKC plays key roles in the meiotic maturation and activation of mammalian oocytes. However, the mechanism of PKC's actions and the PKC isoforms responsible for these actions are poorly understood. In this study, we reveal in mouse eggs and early embryos: (1) the effects of PKC on the meiotic and mitotic cell cycle progression during oocyte maturation, egg activation and embryonic cleavages; (2) the functional importance of classical PKC subclasses in these processes; and (3) the subcellular localization of the PKC alpha isoform during development from GV stage oocytes to the blastocyst stage embryos. The results indicate that the PKC activator phorbol 12-myristate 13-acetate (PMA) inhibits the meiotic resumption of cumulus-free mouse oocytes by a mechanism dependent not only on classical PKC activity but also on other PKC isoforms. PKC activation after germinal vesicle breakdown leads to the inhibition of mitogen-activated protein kinase phosphorylation and the arrest of cell cycle at MI stage. The second polar body emission and the cleavages of early embryos are blocked after prolonged PKC activation. The subcellular localization of PKC alpha isoform in mouse oocytes and embryos is developmental-stage associated. All these results suggest that PKC has multiple functional roles in the cell cycle progression of mouse oocytes and embryos.     

Effects of temperature during early life history on embryonic and larval development and growth in haddock

The effect of incubation temperature (2, 4, 6, 8 and 10° C) on haddock Melanogrammus aeglefinus development and growth during the embryonic period and in subsequent ontogeny in a common post‐hatch thermal environment (6° C) was investigated. Hatching times were inversely proportional to incubation temperature and ranged from 20·3 days at 2° C to 9·1 days at 10° C. Growth rates were directly proportional to incubation temperature during both the embryonic and larval periods. There was a significant decline in growth rates following hatch in all temperature groups. Compared to the endogenously feeding embryos, growth rates in the exogenous period declined by 4·4‐fold at 4° C to 3·9‐fold at 8° C, indicative of the demarcation between the endogenous and exogenous feeding periods. Yolk utilization varied from 17 days at 2° C to 6 days at 10° C and followed a three‐stage sigmoidal pattern with the initial lag period inversely proportional to incubation temperature. Time to 50% yolk depletion varied inversely with temperature but occurred 1–1·5 days post‐hatch at all temperatures. Additionally, the period between 10 and 90% yolk depletion also decreased with increased temperature. Overall developmental rate was sequential with and directly proportional (2·3‐fold increase) to incubation temperature while the time spent in each developmental stage was inversely proportional to temperature. Larger embryos tended to be produced at lower temperatures but this pattern reversed following hatch, as larvae from higher temperature groups grew more rapidly than those from other temperature groups. Larvae from all temperatures achieved a similar length (c.total length 4·5 mm) upon complete yolk absorption. The study demonstrated the significant impact that temperature has upon developmental and growth rates in both endogenous and exogenous feeding periods. It also illustrated that temperature changes during embryogenesis had significant and persistent effects on growth in subsequent ontogeny.     

'Cleavage fields': hypothesis on early embryonic development

The hypothesis is put forward that events of the early embryonic development can be interpreted on the basis of a radial distribution of cytoplasmic components, i.e. of a 'cleavage field', progressively established during the growth of the oocyte. The orientations of the cleavage spindles and the corresponding furrows' positions are assumed to be correlated to the field's temporal evolution which, in turn, is determined by flows of cytoplasmic components originated by the changes in the membrane shape. From this viewpoint, a simple explanation of egg regulation is proposed, and the particular case of the sea urchin embryo is briefly discussed.     

光照强度与光周期对虎斑乌贼胚胎发育的影响

为探究光照对虎斑乌贼受精卵孵化的影响,确定其胚胎发育的最佳光照条件,本研究采用单因子试验方法,分析了不同光照强度(10、30、50、70、90 μmol·m^-2·s^-1)和光周期L∶D(24 h∶0 h、18 h∶6 h、12 h∶12 h、6 h∶18 h、0 h∶24 h)对虎斑乌贼胚胎发育的影响.结果表明: 不同光照强度对虎斑乌贼胚胎发育的孵化率、卵黄囊断裂率、培育周期、初孵幼体体质量与胴长均影响显著;而对孵化周期和幼体出膜7 d后存活率无显著影响.其中孵化率、培育周期、初孵幼体体质量与胴长随着光照强度的增强先增大后减小,而卵黄囊断裂率则逐渐增大.最适光照强度为30 μmol·m^-2·s^-1,此光照强度下孵化率为(90.0±4.1)%,卵黄囊断裂率为(7.3±1.5)%,培育周期为(25.50±0.35) d,孵化周期为(8.10±0.89) d,初孵幼体体质量为(0.213±0.011) g,胴长为(1.013±0.022) cm,出膜7 d后存活率为(97.1±4.0)%.不同光周期对虎斑乌贼胚胎发育的孵化率、培育周期、孵化周期均影响显著,而对卵黄囊断裂率、初孵幼体体质量、胴长和幼体出膜7 d后存活率无显著影响.其中孵化率和孵化周期随着光照时间的增加呈现先增大后减小的变化.最适光周期为LD(12 h12 h),此光周期下孵化率达(88.7±1.8)%,卵黄囊断裂率为(8.7±1.8)%,培育周期为(25.00±0.50) d,孵化周期为(7.00±3.20) d,初孵幼体体质量为(0.209±0.005) g,胴长为(0.998±0.026) cm,出膜7 d后存活率为(96.8±7.1)%.说明弱光照强度30 μmol·m^-2·s^-1和半日光照强度L∶D(12 h∶12 h)更有利于虎斑乌贼的胚胎孵化.在实际生产中,应避免阳光直射,采取适当的遮光措施.     

No influence of the embryonic axis on the development of diamine oxidase in pea cotyledons

A reexamination has been made for the supposed regulation of pea (Pisum sativum cv Alaska) cotyledonary diamine oxidase (EC 1.4.3.6) activity by the embryonic axis. When dry cotyledons from which the embryo and testa have been removed surgically are imbibed by soaking in water, there is little increase of the enzyme activity during subsequent incubation on filter paper. However, if the dry cotyledons are imbibed and maintained on filter paper from the first, the increase of the enzyme activity is similar to that in the intact seedling. Thus, rapid imbibition of the isolated dry cotyledons is responsible for repression of enzyme development, and a role for the axis need not be invoked.     

The expression of c-kit protein during oogenesis and early embryonic development.

The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor and was shown to be allelic with the white-spotting locus (W) of the mouse. Mutations at the W locus have pleiotropic effects on the development of hematopoietic stem cells, melanoblasts, and primordial germ cells. In order to elucidate the role of c-kit protein in gametogenesis and oocyte maturation, we have examined immunohistochemically the expression of c-kit in the ovaries of mice at late fetal and postnatal stages, and in early embryos. By the avidin-biotin-peroxidase (ABC) method using rat anti-mouse c-kit monoclonal antibody, the c-kit protein was detected in ovaries after the time of birth, but not before. The expression of c-kit was observed mainly on the surface of oocytes, but not in granulosa cells nor in interstitial regions. Oocytes of primordial to fully grown Graafian follicles showed the c-kit protein. When ovulation was induced by hCG, the expression of c-kit in ovulated unfertilized oocytes was weaker than in oocytes of Graafian follicles. In 1-cell embryos the c-kit protein was still observed, but with cell division its expression further decreased, and it was not detected in embryos of 4-cell, 8-cell, and morula stages. In summary, the highest expression of c-kit was observed on the surface of oocytes arrested in the diplotene stage of meiotic prophase. With ovulation and the resumption of meiotic maturation, its expression declined. These results suggest that the c-kit protein may play some role in meiotic arrest, oocyte growth, and oocyte maturation.     

Influence of exogenous progesterone on early embryonic development in the mare

The influence of exogenous progesterone on the development of equine oviductal embryos was determined based upon the recovery of Day-7 uterine blastocysts from treated mares (n=13) that were given 450 mg progesterone daily between Days 0 and 6 and from untreated control mares (n=13). Daily administration of 450 mg progesterone in oil significantly (P<0.02) increased serum progesterone concentrations in the treated mares. There was no significant difference in the recovery rate of Day-7 embryos between treated and control mares (8/13 versus 6/13, respectively). Embryonic development, assessed by morphologic evaluation, embryo diameter, and number of cell nuclei was not significantly different for embryos from treated and from control mares. The results of this study indicate that administration of progesterone beginning on the day of ovulation does not affect the embryo recovery rate or embryonic development, based on evaluation of uterine blastocysts recovered at Day 7 after ovulation.     

Effects of EHF radiation and cytoactive substances on fertilization and early embryonic development of the sea urchin Strongylocentrotus intermedius

This study examines the influence of extremely high frequency (EHF) electromagnetic radiation (42.2 GHz, 100 μW/cm², impulse modulation 1000 Hz) on the gametes and embryos of the sea urchin Strongylocentrotus intermedius. Preliminary radiation treatment reduces the number of fertilized egg cells and the survival rate of embryos at early stages of development up to the pluteus. Methyluracil reduces the degree of expression of biological effects of EHF radiation.     

Effects of maternal blood loss on embryonic and placental development in the rat.

The effects of acute loss of maternal blood on embryonic and placental development was examined in 50 rats on Days 8 or 9 of gestation. Blood was withdrawn from conscious, cannulated rats over a 1-min period at 1-0 or 2-0 ml/100 g body weight. These degrees of blood loss were expected to produce a mild (about 50%) and severe (about 80%) reduction in uterine blood flow, respectively, for at least 15 min. There was no evidence that loss of blood affected either fetal survival and malformation rates or fetal weights and sex ratios. The anaemia resulting from haemorrhage lasted no longer than 6 days. Placental weights were 11% higher in rats losing 2-0 ml blood/100 g than in controls (P less than 0-05). It appears that the 8- or 9- day rat embryo is highly resistant to the partial reduction in uterine blood flow, maternal anaemia and other possible challenges induced by maternal loss of blood at levels sufficient to affect the mothers.