An analysis of the inactivation of the frog sperm nucleus by toluidine blue

Toluidine blue, applied to frog sperm under appropriate conditions, inactivates specifically the sperm nucleus, leaving the extranuclear parts of the cell undamaged. Thus, the dye-treated spermatozoa stimulate eggs to cleave normally, but contribute no chromosomes to the resulting embryos, which develop as typical gynogenetic haploids. The concentration of dye required to produce this inactivation varies with pH. Measurements made over the entire pH range which can be tolerated by sperm cells showed that in the lower part of the range (5 to 7) the effective dye concentration was about 5 x 10(-6)M; in the intermediate range (7 to 8.5) it was 1 x (-6) to 1 x (-7)M; and for the higher pH values (8.5 to 10.0) it was about 5 x (-8)M. Using sperm suspensions containing 1500 cells per c. mm. these concentrations of dye produced specific inactivation of the sperm nuclei within 7 to 60 minutes at 18 degrees C. Tests of the reversibility of the inactivation were made by transferring the sperm from the dye to a dilute Ringer's solution after a known degree of inactivation had been produced. Following removal of the dye the sperm cells were tested on eggs over a period of 2 hours. During this time there was no indication of a reversal of the inactivation. Microscopic observations of sperm treated with (-5)M or 5 X (-5)M dye show that the dye is taken up by the sperm nucleus, which is faintly but definitely stained. The dye appears to be uniformly distributed in the nucleus, while extranuclear structures remain unstained. Measurements of the amount of dye bound per sperm nucleus indicate that the minimal quantity required for complete inactivation is about 6.7 x (-18) mole, while the maximal amount which can be bound without injury to extranuclear structures is about 1.5 x (-16) mole. The value obtained for the minimal requirement (6.7 X (16) mole = 4 X (6) molecules) suggests that there are roughly 4 million binding sites in the nucleus which, when blocked by dye molecules, somehow prevent the sperm chromosomes from participating in the development of the egg.     

空肠弯曲菌细胞毒素研究

<正> 空肠弯曲菌是世界性流行胞泻的一个重要原因,许多地方的急性肠炎与之有关,然而人们对其发病机制所知甚少。因为它与炎性胞泻有关,我们推测它可能产生一种细胞毒素,类似于引起炎性腹泻的志贺氏痢疾杆菌或新近出现的引起出白性肠炎的大肠杆菌0.57所产生的毒素。 分离的10株空肠弯曲菌的菌体先经多粘菌素B处理,然后离心过滤得到无细胞滤液,将此滤液稀释后接种到Hela单层细胞(此法比前用于检测志贺氏毒素的细胞毒效应)及新鲜的中国地鼠卵巢单层细胞上,24小时后经苔盼蓝摄取试验确定引起50％细胞死亡的稀释度(CD_(50))。其中5个分离株的10~9细菌/ml处理源滤液CD_(50)为1:4,与Hela细胞与地鼠卵巢细胞结果类似(地鼠