Chicken oocyte growth: receptor-mediated yolk deposition

During the rapid final stage of growth, chicken oocytes take up massive amounts of plasma components and convert them to yolk. The oocyte expresses a receptor that binds both major yolk lipoprotein precursors, vitellogenin (VTG) and very low density lipoprotein (VLDL). In the present study, in vivo transport tracing methodology, isolation of coated vesicles, ligand- and immuno-blotting, and ultrastructural immunocytochemistry were used for the analysis of receptor-mediated yolk formation. The VTG/VLDL receptor was identified in coated profiles in the oocyte periphery, in isolated coated vesicles, and within vesicular compartments both outside and inside membrane-bounded yolk storage organelles (yolk spheres). VLDL particles colocalized with the receptor, as demonstrated by ultrastructural visualization of VLDL-gold following intravenous administration, as well as by immunocytochemical analysis with antibodies to VLDL. Lipoprotein particles were shown to reach the oocyte surface by passage across the basement membrane, which possibly plays an active and selective role in yolk precursor accessibility to the oocyte surface, and through gaps between the follicular granulosa cells. Following delivery of ligands from the plasma membrane into yolk spheres, proteolytic processing of VTG and VLDL by cathepsin D appears to correlate with segregation of receptors and ligands which enter disparate sub-compartments within the yolk spheres. In small, quiescent oocytes, the VTG/VLDL receptor was localized to the central portion of the cell. At onset of the rapid growth phase, it appears that this pre-existing pool of receptors redistributes to the peripheral region, thereby initiating yolk formation. Such a redistribution mechanism would obliterate the need for de novo synthesis of receptors when the oocyte's energy expenditure is to be utilized for plasma membrane synthesis, establishment and maintenance of intracellular topography and yolk formation, and preparation for ovulation.     

鲤鱼鳞被和体色在人工诱导雌核发育中的遗传变异

于1986—1988年,先后人工诱导2尾红镜鲤和4尾荷包红鲤抗寒品系雌核发育,获得雌核发育2倍体当年鱼种2133尾,其中红镜鲤685尾,荷包红鲤1448尾。通过对这些后代鳞被和体色两个质量性状的分析,发现荷包红鲤抗寒品系雌核发育2倍体(1987)中有约三分之一个体表现散鳞型鳞被和桔红色(出现由深到浅一系列变化)及少量桔黄色、肉红色个体。红镜鲤散鳞型鳞被出现体表全部覆盖不规则大型鳞片、2/3、1/2覆盖不规则的大型鳞片和散鳞型个体。体色出现桔红色、桔黄色,上述二色都出现由深到浅一系列变化和肉红色。这两个质量性状在雌核发育代中表现出数量性状的特征。     

伤寒-鼠伤寒重组株Vi4072在小鼠中定居及血清学效果观察试验

将编码Vi抗原的基因克隆到减毒的鼠伤寒沙门氏菌中组建的基因重组株Vi4072,以3×10~8CFU一次口服感染Balb/C小鼠,4天后按7,14,21,28,35,42,49,56,63,70天的间隔收集分离小鼠的集合淋巴结,肝,脾.鉴别是否有本菌出现,并检测血清和小肠匀浆液中的vi抗体.结果表明,感染后49天仍可从脾中分离到该菌;70天仍可从血清和小肠匀浆液中测出vi抗体。     

抗人IgG Fc片段及Fab段单抗的鉴定及应用

本实验采用木瓜酶水解,SPA柱亲合层析等手段得到人IgGFc段及Fab段,以Sigma抗人IgGfFc段和抗人IgG Fab段单抗为标准品,鉴定了细胞库中抗人IgG系列的部分细胞株,得到特异性分泌抗人IgG Fc段和抗人IgG Fab段单抗的细胞各一株。 在上述实验基础上,用抗人IgG Fc及抗人IgG Fab单抗分别制备了Sepharose4B亲合层析柱,提纯了酶解人IgG Fc、Fab片段,经ELISA法鉴定,相互之间无交叉反应。同时用此方法制备了人抗HBe Fab片段,并将该片段进行了过氧化物酶标记,用来配制HBe ELISA诊断盒,证明其生物活性未受影响,而且消除了类风湿因子引起的HBe Ag假阳性现象。因抗HBe单抗来源困难,如采用HBe多抗制备ELISA试剂,本法将是提高质量的一个好方法。     

Restudy of the upper age limit of Beijing Man site

The chronological position of Beijing Man site was restudied by U-series dating of two calcite samples taken from a stalagmitic flowstone layer intercalated in 1–2 layers. Based on the weighted mean of three measurements performed on the purer one, the upper age limit of this site be 421 ₋₅₄ ⁺¹¹⁰ ka B.P., much greater than former evaluation of 230 ka based on U-series dating of fossil bones. The reliability of the obtained results was discussed.     

外显子与内含子序列分维的双碱基研究

引入碱基间的关联,研究了外显子和内含子序列以双碱基为单位的分维,我们发现在这种情况下,外显子和内显子序列在短程和中程存在自相似性并分别定义了这两个区域的分维。结果表明,短程的分维值Ｄｇ一般比中程的Ｄｍ大,外显子的两个分维值比内含子大。我们改变双联体的位相而分维却不变,这反映出在双联体基础上,外显子的不规则性大于内含子,短程的不规则性大于中程,外显子和内含子序列对以２为周期的结构没有位相的特异性。     

Proper phosphorylation of septin 12 regulates septin 4 and soluble adenylyl cyclase expression to induce sperm capacitation

Septin-based ring complexes maintain the sperm annulus. Defective annular structures are observed in the sperm of Sept12- and Sept4-null mice. In addition, sperm capacitation, a process required for proper fertilization, is inhibited in Sept4-null mice, implying that the sperm annulus might play a role in controlling sperm capacitation. Hence, we analyzed sperm capacitation of sperm obtained from SEPT12 Ser196 phosphomimetic (S196E), phosphorylation-deficient (S196A), and SEPT4-depleted mutant mice. Capacitation was reduced in the sperm of both the Sept12 S196E- and Sept12 S196A-knock-in mice. The protein levels of septins, namely, SEPT4 and SEPT12, were upregulated, and these proteins were concentrated in the sperm annulus during capacitation. Importantly, the expression of soluble adenylyl cyclase (sAC), a key enzyme that initiates capacitation, was upregulated, and sAC was recruited to the sperm annulus following capacitation stimulation. We further found that SEPT12, SEPT4, and sAC formed a complex and colocalized to the sperm annulus. Additionally, sAC expression was reduced and disappeared in the annulus of the SEPT12 S196E- and S196A-mutant mouse sperm. In the sperm of the SEPT4-knockout mice, sAC did not localize to the annulus. Thus, our data demonstrate that SEPT12 phosphorylation status and SEPT4 activity jointly regulate sAC protein levels and annular localization to induce sperm capacitation.