Миогенная и нейрональная дифференцировка клеток личинок мидии Mytilus trossulus in vivo и in vitro
Диссертация
Двустворчатые моллюски имеют бифазный жизненный цикл, в котором взрослому животному предшествует стадии свободно плавающих личинок. Очевидное эволюционное преимущество такого цикла — повышенная выживаемость и лучшее расселение потомков. Развитие некоторых двустворчатых моллюсков изучено на уровне электронной микроскопии (Ваупе, 1976; Cragg, 1985; Малахов, Медведева, 1985; Cragg, Crisp, 1991… Читать ещё >
Содержание
- СПИСОК СОКРАЩЕНИЙ
- ГЛАВА 1. ОБЗОР ЛИТЕРАТУРЫ
- 1. 1. Жизненный цикл и миогенез D. melanogaster
- 1. 2. Жизненный цикл и миогенез С. elegans
- 1. 3. Жизненный цикл и миогенез двустворчатых моллюсков
- 1. 4. Анатомия мышц взрослых двустворчатых моллюсков
- 1. 5. Маркеры мышечной дифференцировки
- 1. 6. Анатомия мышц личинок моллюсков
- 1. 7. Нейрогенез личинок моллюсков
- 1. 8. Роль белков, участвующих в сборке толстых филаментов
- 1. 9. Роль эластичных белков в сборке саркомеров
- 1. 10. Роль белков тонких филаментов в сборке саркомеров
- 1. 11. Миофибриллогенез позвоночных животных
- 1. 12. Миогенная и нейрональная дифференцировка клеток беспозвоночных in vitro
- ГЛАВА 3. МАТЕРИАЛ И МЕТОДЫ
- 3. 1. Животные
- 3. 2. Получение актомиозиновых (AM) экстрактов мидии
- 3. 2. 1. Выделение актомиозиновых экстрактов из яйцеклеток и личинок мидии и приготовление проб для электрофореза
- 3. 2. 2. Выделение актомиозиновых экстрактов из переднего аддуктора взрослых животных и приготовление проб для электрофореза
- 3. 3. Электрофорез в 8% полиакриламидном геле в присутствии додецилсульфата натрия (ДСН)
- 3. 3. 1. Приготовление геля
- 3. 3. 2. Условия разделения и окрашивания гелей
- 3. 4. Количественный анализ белков, разделенных электрофорезом
- 3. 5. Получение поликлональных антител кролика к белкам толстых нитей гладких мышц мидии
- 3. 6. Иммуноблотгинг
- 3. 7. Процедура получения первичной культуры клеток мидии
- 3. 7. /. Фракционирование клеток личинок мидии в градиенте Перколла
- 3. 8. Антитела
- 3. 9. Иммунохимия
- 3. 10. Сканирующая конфокальная лазерная микроскопия и обработка изображений
- 3. 12. Определение пролиферативной активности клеток in vitro
- 3. 12. 1. Детекция БДУ в культуре клеток мидии
- 3. 12. 2. Выявление активности щелочной фосфатазы в культуре клеток мидии 44 4.1. Выделение тотальной РНК и ее очистка
- 4. 1. ОТ-ПЦР
- 4. 2. Статистическая обработка полученных данных
- ГЛАВА 4. РЕЗУЛЬТАТЫ
- 4. 1. МИОГЕНЕЗ IN VIVO
- 4. 1. 1. Экспрессия мышечных белков мидии М. trossulus
- 4. 1. 1. 1. Изменение состава мышечных белков в онтогенезе мидии
- 4. 1. 1. 2. Количественная оценка соотношения основных белков мышц личинок и взрослых моллюсков
- 4. 1. 2. Анализ экспрессии твитчина на разных стадиях развития мидии
- 4. 1. 3. Детекция парамиозина в раннем эмбриогенезе мидии
- 4. 1. 4. Морфологический анализ развивающейся мышечной системы
- 4. 1. 5. Локализация мышечных и нейрональных элементов на ранних стадиях развития мидии
- 4. 1. 1. Экспрессия мышечных белков мидии М. trossulus
- 4. 2. МИОГЕНЕЗ IN VITRO
- 4. 2. 1. Первичная культура клеток личинок мидии М. trossulus
- 4. 2. 2. Пролиферация клеток на разных субстратах
- 4. 2. 3. Выявление активности щелочной фосфатазы
- 4. 2. 4. Миофибриллогенез in vitro
- 4. 2. 5. Миогенная и нейрональная дифференцировка в первичной культуре клеток мидии
- 4. 1. МИОГЕНЕЗ IN VIVO
- 4. 1. МИОГЕНЕЗ IN VIVO
- 4. 1. 1. Анализ белкового состава мышц личинок мидии
- 4. 1. 2. Миогенная и нейрональная дифференцировка личинок мидии М. trossulus
- 4. 2. МИОГЕНЕЗ IN VITRO
- 4. 2. 1. Поведение клеток в культуре
- 4. 2. 2. Миофибриллогенез
- 4. 2. 3. Миогенная и нейрональная дифференцировка клеток в культуре мидии
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