Signal transduction pathways in guinea pig sperm

Trifluoperazine (TFP), the antagonist of calmodulin (CaM). significantly stimulated the capacitation and acrosome reaction of guinea pig spermatozoa at the concentration of 10-100μmol/L, independent of the external Ca2+. Forskolin, dbcAMP and caffeine evidently promoted the occurrence of acrosome reaction of spermatozoa at early capacitation stage (5 h) in nonsynchronous system but not in synchronous system. If the spermatozoa were capacitated for 15 h in synchronous system, the above three drugs significantly stimulated acrosome reaction in a Ca2+-independent manner. Protein kinase C activators, i.e. phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-dibutyrate (PDB) did not influence the occurrence of acrosome reaction of spermatozoa at early capacitation stage, but significantly increased the acrosome reaction rate in capacitated spermatozoa in a Ca2+-independent manner. In contrast. PKC inhibitor staurosporine significantly inhibited the occurrence of acrosome reaction.     

兔子宫内膜细胞核雌激素受体的分析

在研究类固醇激素及其受体的相互作用或不同生理、病理条件下激素受体的特性和数量变化的过程中,经常需要制备受体并建立准确可靠的分析方法,以便得到比较理想的结果。为此,我们以兔子宫为材料,分析了内膜细胞核的雌激素受体。一、制备:将屠宰场取来的兔子宫纵向剪开,用冷生理盐水冲洗,解剖刀刮取内膜,按以往粗核方法得到洗过的沉淀,制成0.4MKCI的悬浮液,取一部分测DNA,     

小鼠附睾头精子获得与卵子质膜融合能力的物质基础研究

随着精子在附睾中的转运,它们与卵子质膜的融合能力逐渐增加。怩证明２附睾体和附睾尾的精子均具有相当高的膜融合能力,而附睾头中的精了奶少能与卵子质膜融合,这是还说明附睾头中的精子不具备与云透明带卵子融合的物质条件呢？利用附睾结扎留并延长体外获能时间,可使附睾头远端精子的融合能力明显地提高;在精子培养液中加入ＡＴＰ,并延长精卵共培养时间,也可使一少部分附睾头近端的精子获得与卵子质膜融合的能力。这表明附睾     

文昌鱼受精机理研究——受精卵的超显微结构

青岛文昌鱼受精后受精膜即刻明显举起,皮层颗粒以完整形式胞吐到卵周腔中,分散后大部分皮层颗粒物质与卵黄膜结合一起组成三层结构的受精膜,随着受精膜的举起,它由厚变薄。雄性原核以核膜破裂,染色质去浓缩,扩大重建原核膜形成雄性原核,雌性原核形成是由分离的具有双层膜的染色体联合,膨大而成,雄性原核形成早于雌性原核。雄性原核的迁移受微丝控制。文昌鱼卵黄颗粒中存在线行和环形结构,这种亚显微结构电子密度很高,经实验证明,它既不是微管又不是微丝。     

Immunolocation of antisperm monoclonal antibody 6B10 and corresponding antigen

An antisperm monoclonal antibody 6B10 was produced by hybridoma technique of the isotype IgG. The monoclonal antibody was purified by means of ammonium sulfate precipitation and protein A-Sepharose Cl-4B affinity chromatography. SDS polyacrylamide gel electrophoresis was used to evaluate the purity of the antibody. Evaluation of the sperm acrosomal status was determined by chlortetracycline (CTC) staining. It was found that monoclonal antibody 6B10 can inhibit the sperm acrosome reaction induced by progesterone. The corresponding antigen recognized by monoclonal antibody 6B10 was located on the plasma membrane of the sperm acrosome by indirect immunofluorescent microscopy and immunoelectronmicroscopy. Sperm protein was extracted by 1% Triton X-100. The molecular weight of the antigen is 50 ku, detected by Western blot. The antigen is a key protein in the sperm acrosome reaction and may be the receptor of progesterone on the sperm acrosome. It may either be developed as a candidate contraceptive vaccine     

小鼠球形精子细胞带下受精研究

试验选用电融合法研究小鼠睾丸球形精子细胞带下受精及糖核在卵胞质内的变化规律,结果表明.以DC:3700～3800V/cm,25μs的电场强度,在此前后各维持1MHz,50V/cm的交流电,是球形精子细胞-卵母细胞对(RS-O)融合的理想电场强度,总存活率、融合率和2PN发育率分别为89.0%(575/646),61.9%(356/575)和49.2%(175/356).电融合液中Ca ²⁺浓度对RS-O的存活率、融合率和发育率均无显著影响.注射hCG后14～16h的卵母细胞构建的RS-O的2PN发育率显著高于其它卵龄组.精核在融合后0～4h,其体积无明显变化,致密度逐渐降低,无明显的核仁形成;4～8h后,32.6%(57/175)的精核膨大形成与雌原核相近大小的雄原核;67.4%(118/17)的精核不能充分膨大,形成较小的雄原核,但其可与雌原核结合.192枚受精卵移植后,共产仔鼠12只(世界第2批),产仔率为6.3%(12/192).     

小鼠精子在附睾成熟过程中质膜糖蛋白的变化

利用蛋白印迹法研究了小鼠精子在附睾成熟过程中质膜糖蛋白的变化。结果表明：与花生凝集素结合的１０４ｋＤ和９５ｋＤ糖蛋白,与伴刀豆凝集素结合的１１７、１１４、１０４、８７、７６、７０ｋＤ糖蛋白在睾丸中已完成了蛋白合成及修饰过程;与伴刀豆凝集素结合的１２０ｋＤ和６５ｋＤ糖蛋白,与双花扁豆凝集素结合的５４ｋＤ糖蛋白在精子附睾成熟过程中被修饰成其它种类的蛋白或被丢失。与伴刀豆凝集素结合的１０９ｋＤ糖蛋白,与     

Antigen changes of monoclonal antibody MSH27 in process of post-testicular maturation (in mice)

An anti-mouse spermatozoon monoclonal antibody, MSH27, as well as its purified antigen, can block sperm-egg membrane fusion. As a candidate protein for sperm-egg membrane fusion, the sperm antigen was investigated in the process of post-testicular maturation (PTM). The molecule was produced in testes and located on the plasma membrane of the postacrosomal area of the spermatozoon. However, the epitope recognized by the MSH27 (MSH27Ep) was not exposed until the occurrence of the acrosome reaction. In the process of fertilization, spermatozoa must complete the acrosome reaction before penetrating across the zona pellucidas (ZPs) to approach the plasma membrane of eggs. The effects of the acrosome reaction and penetration of the ZP on the exposure of the MSH27Ep were also studied. It was shown that the percentage of the spermatozoa with the MSH27Ep exposed increased followed with their mature status in PTM. In fact, it bad a linear correlativity with the rate of the acrosome reaction. After spermatozoa had