Variations in the amounts of RNA polymerase forms I, II and III during preimplantation development in the mouse.

DNA-dependent RNA polymerase has been measured at various stages of preimplantation development in mouse embryos. The total RNA polymerase activity per embryo increases rapidly from the 8-cell stage to the blastocyst stage. Studies with low α-amanitin concentrations, which inhibit form II RNA polymerase, and high α-amanitin concentrations, which inhibit both form II and III RNA polymerases indicate that the relative proportions of the three forms change significantly during preimplantation development. The changes which occur in the types and levels of RNA polymerase appear to parallel corresponding changes in the synthesis of the major classes of RNA.     

Quantification of basigin mRNA in mouse oocytes and preimplantation embryos by competitive RT-PCR

Basigin is a member of the immunoglobulin superfamily and a key molecule related to mouse blastocyst implantation. Whether preimplantation mouse embryos express basigin mRNA is still unknown. The aim of this study was to use a quantitative competitive polymerase chain reaction to assess quantitatively the levels of basigin mRNA in mouse oocyte and preimplantation embryos. Basigin mRNA was detected in the oocyte and all the stages of preimplantation embryos. The levels of basigin mRNA were 0.0606 +/- 0.0282 in the oocyte, 0.0102 +/- 0.0036 in the zygote, 0.0007 +/- 0.0003 in the 2-cell embryo, 0.0031 +/- 0.0017 in the 4-cell embryo, 0.0084 +/- 0.0024 in the 8-cell embryo, 0.0537 +/- 0.0121 in the morula and 0.0392 +/- 0.0161 attomoles in the blastocyst, respectively. The levels of basigin mRNA in the oocyte, morula and blastocyst were significantly higher than those in the zygote and embryos at the 2-cell, 4-cell and 8-cell stages. The high level of basigin expression in the blastocyst may play a role during embryo implantation.     

CD63/ME491在植入前小鼠胚胎及延迟着床小鼠子宫的表达

目的 研究肿瘤转移抑制因子CD63/ME491 mRNA和蛋白在植入前小鼠胚胎及延迟着床小鼠子宫中的表达规律,探讨其在胚胎着床过程中的作用以及雌激素对其表达的调节.方法 应用RT-PCR、免疫荧光、免疫组化技术观察CD63/ME491 mRNA和蛋白的表达规律.结果 在植入前小鼠胚胎中均有CD63/ME491 mRNA及其蛋白表达.CD63/ME491 mRNA在桑葚胚及囊胚期表达较丰富,CD63/ME491蛋白表达于各期胚胎细胞的胞膜和胞浆;CD63/ME491 mRNA在延迟着床小鼠子宫均有表达,但从D5到D8呈下降趋势,雌二醇(E2)激活后mRNA的表达显著上升(P<0.05).CD63/ME491蛋白在延迟着床D5弱表达于上皮下基质细胞,D6～8表达不明显,E2激活后该蛋白明显表达于上皮下基质细胞的胞膜和胞浆.结论 1. CD63/ME491在植入前小鼠胚胎中呈动态表达,提示它参与了胚胎的发育过程;2. CD63/ME491在小鼠子宫中的表达可能受雌激素调节.     

Interactions between the cell cycle and embryonic patterning in Arabidopsis uncovered by a mutation in DNA polymerase epsilon

Pattern formation and morphogenesis require coordination of cell division rates and orientations with developmental signals that specify cell fate. A viable mutation in the TILTED1 locus, which encodes the catalytic subunit of DNA polymerase epsilon of Arabidopsis thaliana, causes a lengthening of the cell cycle by approximately 35% throughout embryo development and alters cell type patterning of the hypophyseal lineage in the root, leading to a displacement of the root pole from its normal position on top of the suspensor. Treatment of preglobular and early globular stages, but not later stage, embryos with the DNA polymerase inhibitor aphidicolin leads to a similar phenotype. The results uncover an interaction between the cell cycle and the processes that determine cell fate during plant embryogenesis.     

牦牛LIFR 基因的克隆及其在繁殖周期卵巢中的表达

白血病抑制因子受体(LIFR)与白血病抑制因子(LIF)结合,在排卵、胚胎发育及胚胎附植等过程中起重要的调节作用,与哺乳动物生殖过程密切相关。为进一步研究白血病抑制因子受体(LIFR)基因在卵巢中的表达,本研究对牦牛LIFR 基因进行克隆和序列分析,并利用RT-PCR 技术研究其在繁殖周期卵巢中的表达情况。本研究克隆得到牦牛3 329 bp 的LIFR 基因cDNA 序列,内含3 288 bp 开放阅读框序列。与家牛的相应序列具有很高的同源性(99 5% ),表明LIFR 基因在进化过程中较为保守。实时定量RT-PCR 检测LIFR 基因在繁殖周期卵巢中的表达,结果显示卵巢中LIFR 基因在妊娠期表达最强。表明LIFR 基因在牦牛繁殖周期中具有不可忽视的作用。     

Cloning of total mRNA populations from adult and embryonic mice

Total clone banks of cDNAs synthesized from poly(A)-RNA obtained from three stages of the developing mouse were constructed. The stages chosen were 13-day-old embryo, neonatal, and fully grown adult. To have as complete a bank as possible, large numbers of individual clones were generated ~400,000 for the 13th day embryo and neonatal mouse and ~610,000 for the adult bank. In each case the clone bank was constructed by inserting double stranded cDNA into the PstI site of pBR322 by the “G-C tailing” method. Sequences cloned in this way could be separated from the plasmid host DNA by treatment of the resultant total chimeric plasmid population with PstI. Aliquots of the cloned cDNA material were labeled with ³²P by “nick translation” using Escherichia coli DNA polymerase I for the preparation of hybridization probes. Back-hybridization of these probes to the total clone banks allowed the determination of the sequence diversity among the above three very different developmental stages. The use of such clone banks should allow the identification of developmental stage specific mRNAs.     

达氏鳇不同发育期胚胎对低温的耐受研究

研究了达氏鳇12个发育期胚胎经过不同低温（2 ℃、3 ℃、5 ℃、7 ℃和8 ℃）处理12 h、24 h、2 d、3 d、6 d、10 d、15 d、20 d和30 d后的孵化率和仔鱼成活率.结果表明,卵黄栓期、隙状胚孔期、神经管闭合期胚胎在2～8 ℃水温下,处理24 h后孵化率为0;卵裂期、囊胚早期、原肠中期胚胎在2～8 ℃水温下,处理3 d后孵化率低于30%;囊胚晚期、原肠早期、眼基期、尾芽期、心跳期和尾达头部期胚胎在5～8 ℃水温下,处理3 d后孵化率、仔鱼成活率超过70%;随低温处理时间延长,胚胎和仔鱼的死亡率增加,处理时间与孵化率、仔鱼成活率呈负相关;囊胚晚期、原肠早期、眼基期胚胎在5 ℃水温下耐受力较强,处理10 d后孵化率、仔鱼成活率超过70%.本研究表明,达氏鳇胚胎发育过程中囊胚晚期、原肠早期和眼基期胚胎可以在某一低温下进行短期保存,其孵化率、仔鱼成活率与常温（16～17 ℃）下没有显著差异.这对于达氏鳇胚胎（受精卵）的长途运输有重要意义.     

Chick-embryo DNA polymerase gamma. Identity of gamma-polymerases purified from nuclei and mitochondria

The level of DNA polymerase gamma as compared to DNA polymerases alpha and beta has been determined in chick embryo by means of specific tests: the amount of gamma-polymerase in the 12-day-old chick embryo reaches about 15% of the total polymerase activity. This enzyme is mainly localized in nuclei and mitochondria, where it represents the prevailing if not the unique DNA polymerase activity. The mitochondrial DNA polymerase gamma is likely to be associated with the internal membrane or the matrix of this organelle since it is not removed by digitonin treatment. The gamma-polymerases have been purified from chick embryo nuclei and mitochondria 500-700 times by means of DEAE-cellulose, phosphocellulose and hydroxyapatite chromatographies. The purified mitochondrial DNA polymerase gamma is closely related to the homologous enzyme purified from the nuclei of the same cells. So far, they cannot be distinguished on the basis of their sedimentation, catalytical properties and response to inhibitors or denaturating agents. The purified gamma enzymes are distinct from the chick embryo DNA polymerases alpha and beta and are not inhibited by antibodies prepared against the latter enzymes. The nuclear and mitochondrial gamma-polymerases do not respond to the oncogenic RNA virus DNA polymerase assay with natural mRNAs.     

Antioxidants and total oxyradical scavenging capacity during grass shrimp, Palaemonetes pugio, embryogenesis

During embryogenesis in grass shrimp the capacity to scavenge oxyradicals increased as measured by the Total Oxyradical Scavenging Capacity (TOSC) assay. The increase in TOSC during embryogenesis was associated with increasing concentrations of a number of antioxidants, including coenzyme Q (ubiquinone), alpha-tocopherol and reduced glutathione. Glutathione concentrations ranged from 0.004 to 0.005 nmol/embryo in early embryo stages and reached concentrations between 0.16 to 0.23 nmol/embryo in late embryo stages. Ascorbate remained essentially constant (0.16-0.20 nmol/embryo) throughout embryogenesis and may provide the preponderance of TOSC during early embryo development. Carotenoids were associated with yolk lipovitellin and these antioxidants decreased as yolk was absorbed during embryogenesis. Astaxanthin and beta-carotene were identified in embryos with astaxanthin always the principal carotenoid. In early embryo stages there are maternally derived antioxidants but as embryogenesis proceeds there is an assembly of a complex antioxidant system by newly formed cells and tissues.     

Effect of DNA inhibitors upon DNA synthesis and development of Drosophila embryos

Staged wildtype embryos of Drosophila melanogaster were permeabilized and then subjected to a short pulse of either methyl-3H-thymidine, one of four different inhibitors of DNA synthesis (mitomycin C, 5-fluorouracil, nalidixic acid, or 1-beta-D-arabino-furanosylcytosine-5'-monophosphate), or a combination of both. The incorporation of methyl-3H-thymidine into acid insoluble material was at a maximum during the first half-hour of embryogenesis, after which the incorporation dropped to half the initial value and remained constant throughout the remainder of development. There was no correlation between the rate of incorporation of methyl-3H-thymidine into DNA and the known periods of high mitotic activity. The time course of the estimated specific activity of the DNA newly synthesized in vivo closely paralleled the known changes in the DNA polymerase activity determined in vitro. The known periods of high mitotic activity in the embryo (0-3 hours, 5-12 hours) agree with the periods of maximal sensitivity of the embryo to the inhibitors of DNA synthesis. All four DNA inhibitors affected the incorporation of methyl-3H-thymidine into DNA, although they did not affect it in any simplistic manner. Inhibitor treatment during early cleavage stages resulted in arrested development, treatment during late cleavage and blastoderm stages resulted in abnormal development, and treatment during late blastoderm and early gastrula resulted in normal development. The major phenotypic abnormality caused by the inhibitors is an abnormal distribution of blastoderm cells. As judged by the ID50 values, the embryos remained very sensitive to the effects of the inhibitors until the stages of head and body segmentation, when they then very rapidly became insensitive.     

漆树胚,胚乳发育及花果生长的相关性研究

漆树为倒生胚珠,双珠被,厚珠心,具承珠盘及拟珠孔塞,胚囊发育为蓼型,核型胚乳,胚发育为柳叶菜型,后历经棒状形胚、心形胚、鱼雷形胚和成熟胚各期。花和果实生长与胚及胚乳发育有密切的相关性,胚内具原始的乳汁道系统为重要特征。一些胚珠内无胚或胚乳早期退化引起胚败育是造成种子空籽原因之一。