Salpeter M M, Rogers A W, Kasprzak H, McHenry F A
J Cell Biol. 1978 Jul;78(1):274-85. doi: 10.1083/jcb.78.1.274.
The distribution of acetylcholinesterase (ACHe) in the twitch fibers of the extraocular muscles of the mouse was examined by light and electron microscope autoradiography after labeling with radioactive diisopropyl fluorophosphate (DFP) with, and without, 2-pyridine aldoxime methiodide (2-PAM) reactivation. The values obtained were compared with those previously reported for the diaphragm and sternomastoid muscles. The extraocular muscles were studied because they differ from the other two muscles in that they are among the fastest of the mammalian muscles, yet their endplates have sparse junctional folds. They could thus provide information on the extent to which ACHe concentration is an invariant feature of endplate morphology and what, if any aspects may be related to their fast speed of response. We found, using light microscope autoradiography, that in the twitch fibers of the extraocular muscle, there is n average of 6.4 +/- 2.1 X 10(7) DFP-binding sites per endplate, of which 29% (1.8 X 10(7)) are reactivated by 2-PAM and are thus AChe. The morphology of the extraocular endplates allowed us to conclude, on statistical grounds, that the AChe site are probably localized not only along the surface area of the postjunctional membrane (PJM) but also along the surface of the presynaptic axonal membrane. Based on this localization, we calculate 7,800 DFP sites and 2,500 2-PAM-reactivated sites/micron 2 of surface area of pre-and postjunctional membrane. This stacking density of DFP-binding sites per surface area of membrane ( probably in the overlying sheets of basal lamina) is very similar to that in the diaphragm and sternomastoid muscles.
在用放射性二异丙基氟磷酸酯(DFP)标记后,通过光学显微镜和电子显微镜放射自显影术,对小鼠眼外肌快肌纤维中乙酰胆碱酯酶(ACHe)的分布进行了检查,实验中分别使用和未使用2-吡啶醛肟甲基碘(2-PAM)进行再激活处理。将所得结果与先前报道的膈肌和胸锁乳突肌的结果进行比较。之所以研究眼外肌,是因为它们与其他两块肌肉不同,它们是哺乳动物中收缩最快的肌肉之一,但其终板的连接褶皱稀疏。因此,它们可以提供有关ACHe浓度在多大程度上是终板形态的不变特征的信息,以及哪些方面(如果有的话)可能与其快速反应速度有关。我们通过光学显微镜放射自显影术发现,在眼外肌的快肌纤维中,每个终板平均有6.4±2.1×10⁷个DFP结合位点,其中29%(1.8×10⁷)可被2-PAM再激活,因此是ACHe。眼外肌终板的形态使我们能够基于统计学得出结论,即ACHe位点可能不仅位于突触后膜(PJM)的表面积上,还位于突触前轴突膜的表面。基于这种定位,我们计算出突触前膜和突触后膜每平方微米表面积上有7800个DFP位点和2500个可被二磷醛肟甲基碘再激活的位点。这种每单位膜表面积上DFP结合位点的堆积密度(可能存在于基底层的覆盖片中)与膈肌和胸锁乳突肌中的非常相似。