Полное исчезновение электрического ответа

В 1970 г. мы провели эксперименты, в которых стимулировали нервы, иннервирующие электрические органы, до полного исчезновения электрического ответа. Мы ожидали, что в результате такой обработки должны

заметно снизиться число везикул и содержание в них ацетилхолина. К нашему величайшему удивлению, оказалось, что при стимуляции в первую очередь расходуется и затем возобновляется вневезикулярный ацетилхолин. Напротив, количество везикул и содержание в них ацетилхолина оставались постоянными в течение нескольких минут после стимуляции, и лишь впоследствии везикулярный ацетилхолин начинал истощаться. Для более детального исследования этого явления мы и наши коллеги разработали несколько специальных экспериментальных методов. Чтобы оценивать изменение содержания ацетилхолина в различных компартментах синаптосом за короткие промежутки времени, Израэль и Лесба предложили новую аналитическую методику. Она состоит в следующем. В окружающий синаптосомы раствор вводят смесь ферментов (ацетилхоли- нэстеразы, холииоксидазы и перокси- дазы хрена) и люминофор (люминол). Когда ацетилхолин высвобождается из синаптосом в окружающую среду, он инициирует цепь химических реакций, приводящих к вспышке света, интенсивность которой пропорциональна количеству выделившегося ацетилхолина. Изучая высвобождение ацетилхолина из синаптосом Torpedo, мы пользовались этим тестом.

Синаптосомы трудно стимулировать электрическими импульсами, так что был необходим другой экспериментальный подход. Известно, что если добавить к синаптосомам особые пептиды, называемые ионофорами, то их молекулы образуют в мембранах синаптосом искусственные каналы, через которые могут проходить определенные ионы. Один из таких ионофоров делает мембраны проницаемыми для ионов кальция, а ионы кальция, попадая внутрь синаптосом, вызывают высвобождение ацетилхолина. Количество выделяющегося ацетилхолина может быть измерено описанным выше хемилюминес- центным методом.

 

 

 

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