Acetylcholinesterase activity in the frog neuromuscular junction observed at electron microscopic level after <<vital>> histoenzymatic reaction

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概要

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• The aim of this work was to observe at electron microscopic level acetylcholinesterase (AChE) activity of frog neuromuscular junction revealed by an histoenzymatic reaction made on living tissues. In this purpose, Karnovskys histochemical medium was used for the detection of AChE activity at concentrations ten times more diluted than in the original method. The medium was dissolved in Ringer solution containing citrate buffer, a chelating agent of cupric ions, and additional calcium chloride. Living nerve muscle preparation of the bullfrog was incubated in this histochemical Ringer solution. The muscle activity in response to electrical stimulation of motor nerve (0.1Hz) was monitored by recording of muscular contraction. The incubation was not prolonged beyond 15min, since a blockade of the neuromuscular transmission steadily occurred at that time. The tissue remained alive, however, since transmission was recovered by washing with fresh Ringer solution containing citrate buffer and additional calcium chloride. The primary histochemical precipitates endowed with oxidoreductase-like activity were revealed by secondary incubation with diaminobenzidine (DAB) and hydrogen peroxide. After osmification, osmium black of DAB gave fine localization of AChE activity in the synaptic cleft under the electron microscope.

• 日本学士院の論文

著者

• Tsuji S

  Riken Saitama Jpn

• Tsuji Shigeru

  Institut Des Neurosciences (cnrs Ura 1488) Universite Pierre Et Marie Curie

• Hirai Keiji

  Department Of Autonomic Physiology Medical Research Institute Tokyo Medical And Dental University

• HIRAI Keiji

  Medical Research Institute, Tokyo Medical and Dental University

• MOTELICA-HEINO Ion

  Institut des Neurosciences CNRS URA 1488, Université de Paris VI

• KATAYAMA Yoshifumi

  Medical Research Institute, Tokyo Medical and Dental University

• Motelica‐heino I

  Univ. P. Et M. Curie Paris Fra

• MOTELICA-HEINO Ion

  Institut des Neurosciences CNRS URA 1488, Université de Paris VI

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