Электрофизиологические методы

Электрофизиологические методы дают возможность регистрировать только те процессы, которые происходят на постсинаптической мембране, а активность пресинапти- ческих аксонных окончаний оценивается этими методами лишь косвенно, по действию высвобождающегося из них ацетилхолина.

Между 1959 и 1964 гг. В. Уиттэкер в Кембриджском университете и Э. де Робертис в Университете Буэнос-Айреса вместе со своими коллегами предприняли первые шаги в изучении нервных окончаний прямыми методами. Они показали, что при гомогенизации мозговой ткани грызунов нервные окончания отрываются от аксонов и как бы запаиваются, образуя так называемые синаптосомы. Их можно затем отделить от остальных фрагментов ткани путем центрифугирования в градиенте плотности: в результате центрифугирования синаптосомы собираются в зоне с определенной плотностью. Таким образом, одна из важнейших частей нейрона может быть выделена и проанализирована независимо от остальных компонентов мозга.

Уиттэкер и де Робертис (независимо друг от друга) осуществили и следующий шаг. Они обнаружили, что можно разрушить внешнюю мембрану синаптосом и выделить имеющиеся внутри везикулы. Везикулы, как и ожидалось, действительно содержали ацетилхолин. Но оказалось, что не весь ацетилхолин синаптосом упакован в везикулы — часть его находится непосредственно в цитоплазме. Р. Марчбэнкс из Кембриджского университета тогда предположил, что этот цитоплазматический ацетилхолин также играет важную функциональную роль в проведении нервного импульса.





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