Стресс-реакция у имаго Drosophila: Механизм и генетический контроль тема диссертации и автореферата по ВАК РФ 03.00.15, доктор биологических наук Хлебодарова, Тамара Михайловна

  • Хлебодарова, Тамара Михайловна
  • доктор биологических наукдоктор биологических наук
  • 2000, Новосибирск
  • Специальность ВАК РФ03.00.15
  • Количество страниц 341
Хлебодарова, Тамара Михайловна. Стресс-реакция у имаго Drosophila: Механизм и генетический контроль: дис. доктор биологических наук: 03.00.15 - Генетика. Новосибирск. 2000. 341 с.

Оглавление диссертации доктор биологических наук Хлебодарова, Тамара Михайловна

СПИСОК СОКРАЩЕНИЙ.

ВВЕДЕНИЕ.

ОБЗОР ЛИТЕРАТУРЫ.

Ювенильный гормон./

Регуляция титра ЮГ.

Система деградации ЮГ.

Механизм действия ЮГ.

Роль ЮГ в размножении насекомых.

Развитие насекомых в неблагоприятных условиях среды.

Реакция системы деградации ЮГ на стресс.

Механизм стресс-реакции у личинок насекомых и его генетический контроль.

Биогенные амины насекомых./

Влияние стресса на содержание биогенных аминов у насекомых.

Система метаболизма дофамина и октопамина у ЭгоБорЬПа.

Щелочная фосфатаза.

Тирозин гидроксилаза.

Фенолоксидазы.

Дофадекарбоксилаза.

Тирозиндекарбоксилаза.

Ы-ацетилтрансфераза.

И-Р-аланилдофаминсинтетаза.

Дофамин-Р-гидроксилаза.

Внутриклеточный стресс-ответ.

Белки теплового шока и их функции в клетке.

Молекулярные механизмы регуляции экспрессии БТШ.

Рекомендованный список диссертаций по специальности «Генетика», 03.00.15 шифр ВАК

Заключение диссертации по теме «Генетика», Хлебодарова, Тамара Михайловна

ВЫВОДЫ.

1. Показано, что у самок И. и D. melanogaster система деградации ювенильного гормона участвует в реализации стрессорного ответа. Она играет ключевую роль в регуляции размножения, контролируя откладку яиц. Механизм этого контроля связан с сайт-специфической регуляцией титра ювенильного гормона, который обеспечивается у разных видов насекомых различными ЮГ-гидролизующими ферментами, в частности, у 1). утШ это ЮГ-эстераза, у молодых самок А melanogaster -ЮГ-эстераза и ЮГ-эпоксидгидраза, а у половозрелых - только ЮГ-эпоксидгидраза.

2. Установлено, что у самцов I). утШ и О. melanogaster система деградации ювенильного гормона не участвует в реализации стрессорного ответа и не играет существенной роли в их репродуктивной функции.

3. Выяснено, что у имаго И. \inlis повышение уровня биогенных аминов в начале развития стрессорной реакции связано, по-видимому, с их выбросом из депо, а последующее поддержание их высокого уровня определяется активностью ферментов их синтеза: щелочной фосфатазы, контролирующей пул тирозина, тирозингидроксилазы и тирозиндекарбоксилазы, конвертирующих тирозин в ДОФА и тирамин, соответственно

4. Обнаружено, что у имаго мутантной линии 147 О. утШ нарушены механизмы регуляции активности ферментов метаболизма ювенильного гормона и биогенных аминов, которые у ОгозорЫ1а участвуют в реализации стрессорного ответа, а именно: ЮГ-эстеразы, тирозингидроксилазы, тирозиндекарбоксилазы и щелочной фосфатазы.

5. Генетический анализ выявленных межлинейных различий в уровне активности ДФФ-чувствительной и ДФФ-нечувствительной ЮГ-эстераз, щелочной фосфатазы и тирозиндекарбоксилазы показал, что каждое из них контролируется моногенно. Гены, контролирующие активность ЩФ и ТДК были локализованы в одной хромосоме -хромосоме 6 В. \irilis. Гены, контролирующие стресс-реактивность системы дофамина и ответ системы деградации ЮГ на стресс у В. у/п/и также были локализованы в хромосоме 6.

6. Получены свидетельства того, что контроль двух важнейших защитных реакций - нейрогормональной стрессорной реакции и реакции теплового шока - у БгозорЫк осуществляется через независимые сигнальные пути, поскольку нарушение одной из них не препятствует развитию другой.

7. На основании полученных данных сформулировано представление о существовании единого генетического контроля изученных звеньев нейрогормональной стрессорной реакции: ювенильного гормона и биогенных аминов.

8. Из сопоставления фенотипических особенностей мутантной линии 147 В. уМШ и свойств генов, расположенных в хромосоме 4 В. те1ап(^а81ег (структурном аналоге хромосомы 6 В. утШ), следует, что геном, участвующим в контроле нейрогормональной стрессорной реакции у ОгозорМа, может быть ген рибосомного белка ЯрБЗа, обладающий множественными, в том числе и экстрарибосомными, функциями.

ЗАКЛЮЧЕНИЕ.

Итак, в настоящей работе впервые проведено исследование системы деградации ювенильного гормона и системы метаболизма дофамина и октопамина у имаго БгоБорНИа утШ, которые, как показано нами, участвуют в реализации стрессорного ответа и находятся под общим генетическим контролем.

Анализ полученных результатов свидетельствует о том, что система деградации ЮГ, которая у разных видов насекомых может обеспечиваться разными ферментами, ЭГ-эстеразой и/или ЮГ-эпоксигидразой, контролирует откладку яиц и играет, таким образом, ключевую роль в регуляции размножения насекомых. В то же время, наше исследование показало, что ювенильный гормон и, в том числе, система его деградации не играют существенной роли в репродуктивной функции самцов Бго8орЫ1а. Отсутствие в литературе каких-либо данных об участии ЮГ в регуляции репродуктивных функций у самцов других видов насекомых позволяет предположить, что эта характеристика не является особенностью Ого5орЫ1а.

Анализ системы метаболизма ДА и ОА у имаго БгозорЬйа показал, что повышение уровня биогенных аминов при стрессе у насекомых вызвано их выбросом из депо, а поддержание их высокого уровня в условиях длительного стресса связано с инактивацией системы их депонирования и последующим повышением уровня активности ферментов, контролирующих начальные этапы их синтеза.

Анализ развития стрессорной реакции на уровне клетки показал, что она комплексная и отдельные ее звенья могут контролироваться через независимые механизмы, как это мы показали для "хит-шок" ответа клетки, который, являясь одним из ее элементов, индуцируется хотя и одновременно, но независимо и не является пусковым звеном стрессорной реакции, развивающейся на уровне целого организма.

Полученные результаты позволили нам выдвинуть гипотезу о природе гена, мутация которого нарушила у D. virilis стресс-реактивность систем метаболизма ЮГ и ДА. Тот факт, что эта мутация нарушила не только ответ на стресс изученных систем, но и снизила эффективность трансляции белков, позволила нам предположить, что этот ген обладает множественными функциями в организме, а локализация его в хромосоме 6 D. virilis, которая является мини-хромосомой и содержит ограниченный набор генов, позволила предположить, что таким геном может быть ген рибосомного белка RpS3a. Как показано, этот белок входит в состав инициаторного комплекса, контролируя таким образом эффективность трансляции, участвует в регуляции клеточного цикла, репродуктивной функции, апоптоза, онкогенной трансформации и в ответе на активацию TNFa, что свидетельствует о его множественных функциях в организме.

В целом, представленные результаты свидетельствуют о том, что в генерализованном ответе на стрессорное воздействие у Drosophila участвует не одна сигнальная система, как на уровне клетки, где показана независимая индукция синтеза белков теплового шока, так и на уровне целого организма, о чем свидетельствуют полученные ранее данные, что у линии 147 D. virilis не нарушена реакция на стрессорное воздействие нейросекреторных клеток мозга, контролирующих синтез и секрецию проторакотропного гормона (Раушенбах, Лукашина, 1983; 1984; Rauschenbach etal., 1987; Раушенбах, 1990).

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