Biotin-containing proteins of the insect nervous system, a potential source of interference with immunocytochemical localization procedures.
Biotin-containing proteins of the insect nervous system, a potential source of interference with immunocytochemical localization procedures.
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昆虫神经系统中含有生物素的蛋白质，是免疫细胞化学定位程序干扰的潜在来源。
DOI：
10.1016/0965-1748(94)00095-y
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发表时间：
1995
影响因子：
3.8
通讯作者：
Davis,NT
中科院分区：
文献类型：
--
作者：
Ziegler,R;Engler,DL;Davis,NT
当生物素化的天南星脂肪动力激素基因被用作原位杂交的探针时，用含有链霉亲和素或亲和素的生物素检测系统对内在神经分泌细胞进行染色。进一步的实验表明，DNA探针是不必要的染色这些细胞的链霉亲和素-碱性磷酸酶，他们没有染色的碱性磷酸酶单独。同样，内在神经分泌细胞直接染色的链霉亲和素共轭的荧光染料。中枢神经系统的其他部分也可以用链霉亲和素-碱性磷酸酶染色，但不像心体的内在神经分泌细胞那样容易。进一步的分析表明，三个含生物素的蛋白质在心体和大脑的内在神经分泌细胞。当通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析时，发现这些蛋白质中最丰富的蛋白质具有130，000的分子量，这是丙酮酸羧化酶（一种含生物素的酶）亚基的大小。相同的蛋白质被抗昆虫丙酮酸羧化酶的抗血清识别，表明该蛋白质可能是丙酮酸羧化酶。本文报道的结果表明，心体的内源性神经分泌细胞可能含有丙酮酸羧化酶的浓度高于中枢神经系统的其他细胞。我们还注意到，如果使用含有抗生物素蛋白的检测系统进行原位杂交或免疫细胞化学，则需要谨慎以避免假阳性结果。
When the biotinylated Manduca sexta adipokinetic hormone gene was used as a probe for in situ hybridization, the intrinsic neurosecretory cells were stained with a biotin detection system that contained streptavidin or avidin. Further experiments showed that the DNA probe was not necessary for staining these cells by streptavidin-alkaline phosphatase, and that they were not stained by alkaline phosphatase alone. Similarly, the intrinsic neurosecretory cells were stained directly by streptavidin conjugated to a fluorescent dye. Other parts of the central nervous system could also be stained with streptavidin-alkaline phosphatase but not as readily as the intrinsic neurosecretory cells of the corpora cardiaca. Further analysis demonstrated three biotin-containing proteins in the intrinsic neurosecretory cells of the corpora cardiaca and in the brain. The most abundant of these proteins, when analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, was found to have a molecular weight of 130,000, which is the size of the subunits of pyruvate carboxylase, a biotin-containing enzyme. The same protein was recognized by an antiserum against an insect pyruvate carboxylase, indicating that this protein is probably pyruvate carboxylase. The results reported here indicate that the intrinsic neurosecretory cells of the corpora cardiaca may contain pyruvate carboxylase in a concentration higher that other cells of the central nervous system. We also note that caution is necessary to avoid false positive results if an avidin containing detection system is used for in situ hybridization or immunocytochemistry.