成年小鼠大脑中鼠神经生长因子（mNGF）的生物测定检测

Bioassay detection of mouse nerve growth factor (mNGF) in the brain of adult mice.

作者信息

Scott S M, Tarris R, Eveleth D, Mansfield H, Weichsel M E, Fisher D A

出版信息

J Neurosci Res. 1981;6(5):653-8. doi: 10.1002/jnr.490060510.

Abstract

Two pools of seven brains each from adult Swiss-Webster mice were homogenized, and supernatants were collected for bioassay. PC-12 cells were placed in a bioassay plate at time zero, at a concentration of 10(4) cells per well, and primed for 48 hours in a medium containing 50 ng/ml of mNGF. The PC-12 cell bioassay for neurite outgrowth was conducted after primed cells were exposed to an NGF-free medium for 24 hours. Suitable controls for serum toxicity and cell viability were established. The sensitivity of the bioassay approximates 100 pg NGF/ml. The results showed 80-100% neurite outgrowth in wells exposed to brain pool supernatant (BPS) alone, and control level outgrowth (3-8%) in wells containing BPS and specific anti-beta-NGF antibody. Therefore, the brains of Swiss-Webster adult mice contain an NGF-like substance which promotes neurite outgrowth in PC-12 cells. The substance probably is NGF itself, since the effect is blocked by specific NGF antiserum.

摘要

将来自成年瑞士 Webster 小鼠的两组，每组七个大脑进行匀浆，并收集上清液用于生物测定。在时间为零时，将 PC - 12 细胞以每孔 10(4) 个细胞的浓度接种到生物测定板中，并在含有 50 ng/ml 小鼠神经生长因子（mNGF）的培养基中预孵育 48 小时。在预孵育的细胞暴露于无 NGF 培养基 24 小时后，进行 PC - 12 细胞神经突生长的生物测定。建立了血清毒性和细胞活力的合适对照。该生物测定的灵敏度约为 100 pg NGF/ml。结果显示，仅暴露于脑池上清液（BPS）的孔中有 80 - 100% 的神经突生长，而含有 BPS 和特异性抗β - NGF 抗体的孔中神经突生长为对照水平（3 - 8%）。因此，成年瑞士 Webster 小鼠的大脑中含有一种促进 PC - 12 细胞神经突生长的 NGF 样物质。该物质可能就是 NGF 本身，因为其作用被特异性 NGF 抗血清所阻断。