Specific binding of three neurotoxins with phospholipase A2 activity to synaptosomal membrane preparations from the guinea pig brain

Snake presynaptic toxins such as crotoxin, β-bungarotoxin and taipoxin block neuromuscular transmission through inhibiting the release of acetylcholine by their phospholipase A₂ activities. On the other hand, many other phospholipase A₂s show little neurotoxicity. It is likely that the difference lies in whether high affinity binding to nerve cell membranes exists or not. To test this idea, crotoxin, β-bungarotoxin and taipoxin were first radioactively labeled with Na(¹²⁵I) without loss of their neurotoxicity. Using the radioactive toxins we have found that each of the three showed specific binding to synaptosomal membranes from guinea pig brain. In contrast, we could not detect specific binding of a non-neurotoxic pancreatic phospholipase A₂. Crotoxin and taipoxin, but not β-bungarotoxin, also bound specifically to membrane preparation from other tissues. The binding of each toxin was not greatly affected by the other two toxins. The photoaffinity labeling technique has been used to obtain further information about the components which bind crotoxin. For this purpose, (¹²⁵I) crotoxin was derivatized with N-hydroxysuccinimidyl-4-azidobenzoate. Autoradiographic analysis of the membranes following photoirradiation in the presence of the modified crotoxin revealed that an 85K dalton component was preferentially covalently conjugated with the crotoxin analogue in a specific manner.     

利用组织培养选择烟草耐盐愈伤组织变异体并分化出再生植株

烟草(品种革新一号)叶片为外植体,直接置入含0.5%NaCl的修改MS培养基中,诱发产生耐盐的愈伤组织。然后采取逐步提高NaCl浓度的措施,分别获得耐0.5%、1.0%、1.5%及2.0%NaCl细胞系。耐盐细胞系在无盐条件下,生长9—11代后仍保持其耐盐性。从各个耐盐细胞系均分别获得再生苗。耐2.0%NaCl的04—9细胞系共得到15株再生植株,叶片狭长、多锯齿、并具有较多的花茎,多数花粉粒畸形,经过人工授粉获得少量种子。04-9变异型再生植株水培于含有1.0—2.0%NaCl的Hogland溶液中生长85天,仍然存活。原始型愈伤组织的细胞呈不规则椭圆形,耐盐细胞系的细胞均近似圆形;耐盐浓度愈高则细胞愈小。耐盐细胞系愈伤组织的叶绿素含量随耐盐浓度增高而增加;渗透势则随耐盐水平提高而降低。耐2.0%NaCl细胞系04—9愈伤组织内脯氨酸含量高40.7倍,其再生植株叶片内的脯氨酸含量亦较原始型增加两倍。耐2.0%NaCl细胞系再生植株的幼年与成年叶片的过氧化物同工酶的酶谱与原始型均有显著差别。以上试验结果均说明耐2.0%NaCl细胞系04—9及其再生植株是一个耐盐变异体。     

外胚层细胞诱导后的间隙连接

用电镜观察了经受诱导作用之后胚胎细胞的冷冻蚀刻复型膜。和未经诱导作用的对照组比较,早期和中期神经胚的神经上皮细胞以及经过豚鼠骨髓粗提液(BME)——一种有效的异源中胚层诱导物——处理过的早期原肠胚外胚层,它们的间隙连接都处于活跃的动态状态。用图像分析仪测得的间隙连接连接子的排列密度,指出经受过诱导作用的三组分别和未经受诱导作用的对照组比较,计算求出P值,神经上皮两组和对照组的差别为非常显著,BME处理过的细胞和对照组的差别为显著。结合对照组与诱导后胚胎细胞间隙连接连接子的变化讨论了它们在信息传递上可能起的作用。     

An essential tryptophan residue for rabbit muscle creatine kinase

The tryptophan residues in rabbit muscle creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2) have been modified by dimethyl(2-hydroxy-5-nitrobenzyl) sulfonium bromide after reversible protection of the reactive SH groups. The modification of two tryptophan residues as measured by spectrophotometric titration leads to complete loss of enzymatic activity. Control experiments show that reversible protection of the reactive SH groups as S-sulfonates followed by reduction results in nearly quantitative recovery of enzyme activity. The presence of a 410 nm absorption maximum and the decrease in fluorescence of the modified enzyme indicate the modification of tryptophan residues. At the same time, SH determinations after reduction of the modified enzyme show that the reagent has not affected the protected SH groups. Quantitative treatment of the data (Tsou, C.-L. (1962) Sci. Sin. 11, 1535 1558) shows that among the tryptophan residues modified, one is essential for its catalytic activity. The presence of substrates partially protects the modification of tryptophan residues as well as the inactivation, suggesting that the essential tryptophan residue is situated at the active site of this enzyme.     

Regional Distribution of Calmodulin Activity in Rat Brain

Calmodulin activity in 68 discrete areas of rat brain, obtained by micropunch technique, was assessed by its capacity to activate a calmodulin-sensitive form of phosphodiesterase. In general, the activity of calmodulin was higher in the telencephalon, limbic system, and hypothalamus than in the mesencephalon, pons, cerebellum, and medulla. However, there were substantial differences in calmodulin activity in discrete nuclei of each region. The regional distribution of calmodulin activity in rat brain does not appear to correlate with that of any of the known putative neurotransmitters or peptides.     

金沙江河谷苏铁天然植物群落的研究

近年在金沙江干热河谷及其邻近地区陆续发现的苏铁天然植物群落,为本属植物在欧亚大陆内陆分布的最北边缘。其中,面积最大和保存较好的是渡口市攀枝花苏铁(Cycas panzhihuaensis)植物群落。本文研究了这个群落的分布与生境、生长状况、外貌、种类组成以及群落的综合特征等。认为具有稀树草原的特点,而这里的稀树草原是次生性质的,苏铁群落是其中幸存的残遗植被类型。这对研究我国植物地理有着重要意义。     

向日葵未受精胚珠培养时胚状体发生的显微观察

由开花前1—4天的向日葵子房中取出胚珠,在液体培养基上进行漂浮培养,诱导了未受精的卵细胞发育为单倍体的胚状体.亦诱导了珠被绒毡层产生胚状体。对两种胚状体的发生和发育过程及其形态发生特点作了显微观察与描述。     

成人腹泻轮状病毒的提纯及其高价免疫血清的制备

本文采用交叉免疫电泳技术,首先提纯了成人腹泻轮状病毒(Adult DiarrhoeaRotavirus简称:ADRV),并经电镜确认,用作免疫抗原,首次制备了高价兔与豚鼠抗ADRV血清和小鼠抗ADRV腹水,其特异性经交叉免疫电泳和对流免疫电泳鉴定,其效价经酶联免疫吸附试验(ELISA)检测均在1:2,000以上。     

Human chromosome hot points

Summary Peripheral lymphocytes from 16 healthy adults, 9 pregnant women, and 3 fragile X syndrome patients were cultured in Eagle's minimum essential medium without folic acid (MEM-FA). The addition of 2mM, 4mM, or 8mM uridine 24h or 72h prior to harvest resulted in increases of chromosome gaps or breaks, especially at hot points 3p14, 16q23-24, and at fragile site Xq27. Pregnant women showed higher frequencies of 3p14 breaks and total chromosome breaks than men and non-pregnant women. The other chromosome regions, such as 6q26, 7q23, 7q35, 6p25, Xp22, 14q23 and 11p13, also frequently showed gaps or breaks. The results indicated that the unbalance of nucleotide pools was one of the causes of chromosome breakages. The higher frequencies of chromosome gaps and breaks under the condition of thymidylate stress may be due to the misincorporation of uracil instead of thymine into DNA.