Malina Tamás, Krecsák László, Westerström Alexander, Szemán-Nagy Gábor, Gyémánt Gyöngyi, M-Hamvas Márta, Rowan Edward G, Harvey Alan L, Warrell David A, Pál Balázs, Rusznák Zoltán, Vasas Gábor
Pfizer Hungary Ltd., Medical Division, Alkotás u. 53, H-1123, Budapest, Hungary.
Hattyú u. 1, H-1015, Budapest, Hungary.
Toxicon. 2017 Sep 1;135:59-70. doi: 10.1016/j.toxicon.2017.06.004. Epub 2017 Jun 9.
We have revealed intra-population variability among venom samples from several individual European adders (Vipera berus berus) within a defined population in Eastern Hungary. Individual differences in venom pattern were noticed, both gender-specific and age-related, by one-dimensional electrophoresis. Gelatin zymography demonstrated that these individual venoms have different degradation profiles indicating varying protease activity in the specimens from adders of different ages and genders. Some specimens shared a conserved region of substrate degradation, while others had lower or extremely low protease activity. Phospholipase A activity of venoms was similar but not identical. Interspecimen diversity of the venom phospholipase A-spectra (based on the components' molecular masses) was detected by MALDI-TOF MS. The lethal toxicity of venoms (LD) also showed differences among individual snakes. Extracted venom samples had varying neuromuscular paralysing effect on chick biventer cervicis nerve-muscle preparations. The paralysing effect of venom was lost when calcium in the physiological salt solution was replaced by strontium; indicating that the block of twitch responses to nerve stimulation is associated with the activity of a phospholipase-dependent neurotoxin. In contrast to the studied V. b. berus venoms from different geographical regions so far, this is the first V. b. berus population discovered to have predominantly neurotoxic neuromuscular activity. The relevance of varying venom yields is also discussed. This study demonstrates that individual venom variation among V. b. berus living in particular area of Eastern Hungary might contribute to a wider range of clinical manifestations of V. b. berus envenoming than elsewhere in Europe.
我们揭示了匈牙利东部一个特定种群中,几只欧洲蝰蛇(Vipera berus berus)毒液样本的种群内变异性。通过一维电泳发现了毒液模式的个体差异,包括性别特异性和年龄相关性。明胶酶谱分析表明,这些个体毒液具有不同的降解谱,表明不同年龄和性别的蝰蛇标本中蛋白酶活性不同。一些标本共享底物降解的保守区域,而其他标本的蛋白酶活性较低或极低。毒液的磷脂酶A活性相似但不完全相同。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)检测到毒液磷脂酶A谱(基于成分的分子量)的标本间多样性。毒液的致死毒性(LD)在个体蛇之间也存在差异。提取的毒液样本对鸡颈二腹肌神经-肌肉制剂具有不同的神经肌肉麻痹作用。当生理盐溶液中的钙被锶取代时,毒液的麻痹作用消失;这表明对神经刺激的抽搐反应阻滞与磷脂酶依赖性神经毒素的活性有关。与迄今为止研究的来自不同地理区域的贝氏蝰蛇毒液不同,这是首次发现主要具有神经毒性神经肌肉活性的贝氏蝰蛇种群。还讨论了毒液产量变化的相关性。这项研究表明,生活在匈牙利东部特定地区的贝氏蝰蛇个体毒液变异,可能导致贝氏蝰蛇咬伤的临床表现范围比欧洲其他地方更广。