Результаты опытов с синаптосомами

Результаты наших опытов с синаптосомами согласовались с данными, полученными ранее на интактной ткани: первым из синаптосом высвобождался цитоплазматический ацетилхолин. Даже если мы лишали синаптосомы ббльшей части их везикул и затем повторно заполняли экзогенным ацетилхолином, кальциевая стимуляция приводила к высвобождению ацетилхолина. Следовательно, присутствие везикулярного ацетилхолина не обязательно для процесса высвобождения ацетилхолина из синаптосом.

Нам казалось важным провести быстрый биохимический анализ интактной ткани во время высвобождения в ней ацетилхолина. С этой целью мы сконструировали в Женевском университете специальную установку, позволяющую стимулировать несколько выделенных из электрического органа столбиков в быстрой последовательности (см. рисунок на с. 30). При такой стимуляции к одному и тому же моменту времени столбики достигали разных стадий в процессе высвобождения ацетилхолина, которое можно было резко останавливать, быстро охлаждая препараты.

В одном из экспериментов мы проводили стимуляцию короткими сериями импульсов каждые 10 мс в течение 200 мс. Такой характер стимуляции был выбран потому, что он близок к естественному возбуждению в электрическом органе Torpedo. Опять, и на этот раз уже без особого удивления, мы обнаружили, что содержание ацетилхолина в синаптических везикулах при стимуляции не меняется.

Более того, оказалось, что количество вневезикулярного ацетилхолина снижается в течение первых 100 мс стимуляции, на протяжении следующих 100 мс оно увеличивается, а затем вновь снижается. Образование ацетилхолина в интактных нервных окончаниях, несомненно, представляет собой чрезвычайно быстрый процесс. То, что содержание ацетилхолина в конце концов снижается, обусловлено, видимо, тем, что нервные окончания не успевают синтезировать ацетилхолин в количестве, достаточном, чтобы скомпенсировать его высвобождение.

БЫЛО трудно поверить, что ацетилхолин может проходить через мембрану окончания аксона с высокой скоростью, не вызывая пусть временных, но все же доступных для наблюдения структурных изменений. Если бы синаптические везикулы принимали непосредственное участие в высвобождении ацетилхолина, то мы бы могли увидеть их слияние с мембраной нервного окончания и открывание в синаптическую щель, что должно было бы происходить при каждом акте высвобождения ацетилхолина из нейрона.

К настоящему времени накопилось множество электронных микрофотографий, на которых видно слияние везикул с мембраной нервного окончания в нервно-мышечных синапсах. Такое слияние, однако, не всегда наблюдается в тех условиях, когда происходит высвобождение ацетилхолина. Мы получили несколько электронных микрофотографий нервных окончаний электрического .

 

 

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