Analysis of the expression levels of genes that encode cytoskeletal proteins in Drosophila melanogaster larvae during micro- and hypergravity effect simulations of different durations
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Огнева И. В.
Свистунов А.А
Несвижский Юрий Владимирович
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Biophysics (Russian Federation) |
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The goal of this study was to find genes that encode cytoskeletal proteins that are potential candidates for the role of triggers in cell mechanosensitivity in the fruit fly. Centrifugation was used to simulate the hypergravity effects (2g group); the constantly changing orientation of the larvae in the gravity field was performed in order to simulate the effects of microgravity (0g group) for 1.5, 6, 12 and 24 h. mRNA levels of different genes that encode the components of both tubulin and actin cytoskeleton were assessed by qRT-PCR. In the 0g group the mRNA levels of beta-tubulin and Msps were reduced after 1.5 h of the exposure and remained unchanged until 12 h, while they exceeded the control level after 24 h. The mRNA level of chaperonin containing T-complex 1 polypeptide subunits recovered earlier: after 6 and 12 h of the microgravity exposure. At the same time, the hypergravity effect led to more significant changes in the mRNA level of TCP1 complex components compared with those of tubulin and Msps. The mRNA level of beta-actin isoforms under micro- and hypergravity was decreased up to 12 h of the exposure, however, it remained reduced under microgravity conditions, while it recovered (Act87E) and even exceeded (Act57B) the reference level under hypergravity conditions. The mRNA level of supervillin was almost unchanged. Under microgravity conditions the mRNA level of fimbrin was decreased (it recovered by the 24 h time point), while the mRNA level of alpha-actinin was significantly increased by the 12 h time point of the exposure and after 24 h it was reduced to the control level. In contrast, under hypergravity conditions the mRNA level of fimbrin initially increased, and after 24 h it dropped below the control, while the mRNA level of alpha-actinin was significantly reduced, and after 24 h it was higher than the reference level. Similar results were obtained earlier in the experiments in rodents, but similar dynamics were observed for alpha-actinin isoforms 1 and 4, although no changes were observed for fimbrin. Since Drosophila melanogaster has no alpha-actinin isoform 4, it is hypothesized that its role in the cell is played by fimbrin.
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Влияние введения смеси фосфотидилхолинов на состояние кортикального цитоскелета волокон камбаловидной мышцы крысы
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Огнева И. В.
Свистунов А.А.
Несвижский Юрий Владимирович
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Авиакосмическая и экологическая медицина |
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Цель работы - оценка состояния кортикального цитоскелета волокон камбаловидной мышцы крысы в результате 6-часового антиортостатического вывешивания на фоне предшествующего, в течение 3 сут по 100 мкг/сут, введения смеси фосфатидилхолинов (лецитина). Данные о периметре волокон, толщине подмембранного цитоскелета и доле разрывов в нем относительно периметра получали, используя иммуногистохимическую окраску на альфа-актинин-4. Периметр волокон оставался неизменным во всех группах исследования. При этом доля разрывов была выше в группах вывешивания, чем в соответствующих контрольных группах. Толщина окрашенного слоя, соответствующего подмембранному цитоскелету, не менялась в контрольных группах и в группе вывешивания без лецитина, однако в группе вывешивания на фоне лецитина она достоверно увеличивалась по сравнению с соответствующей контрольной группой на 27 % (p < 0,05) соответственно.
Purpose of the work was to assess the cortical cytoskeleton of m. soleus fibers from rats after 6-hr tail-suspension preceded by 3 days of phosphatidylcholine (lecithin) injections at a dose of 100 µг/d. Data about the fiber perimeter, submembrane cytoskeleton thickness and percentage of bonds breaks along the perimeter were obtained using the alpha-actinin-4 anti-body stain. The fiber perimeter remained unchanged in all groups under study. However, the percentage of breaks was high in suspension groups but not in respective groups of control. Thickness of the stained layer commensurate to the submembrane cytoskeleton did not change in the control groups and in the suspension group without lecithin injections but increased reliably in the suspension group that recieved injections by 27 % (p < 0.05) in comparison with its control.
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Публикация |
Analysis of the expression levels of genes that encode cytoskeletal proteins in Drosophila melanogaster larvae during micro- and hypergravity effect simulations of different durations
|
|
Огнева И. В. (Профессор)
Свистунов А.А (Первый проректор)
Несвижский Юрий Владимирович (Профессор)
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Biophysics (Russian Federation) |
|
|
The goal of this study was to find genes that encode cytoskeletal proteins that are potential candidates for the role of triggers in cell mechanosensitivity in the fruit fly. Centrifugation was used to simulate the hypergravity effects (2g group); the constantly changing orientation of the larvae in the gravity field was performed in order to simulate the effects of microgravity (0g group) for 1.5, 6, 12 and 24 h. mRNA levels of different genes that encode the components of both tubulin and actin cytoskeleton were assessed by qRT-PCR. In the 0g group the mRNA levels of beta-tubulin and Msps were reduced after 1.5 h of the exposure and remained unchanged until 12 h, while they exceeded the control level after 24 h. The mRNA level of chaperonin containing T-complex 1 polypeptide subunits recovered earlier: after 6 and 12 h of the microgravity exposure. At the same time, the hypergravity effect led to more significant changes in the mRNA level of TCP1 complex components compared with those of tubulin and Msps. The mRNA level of beta-actin isoforms under micro- and hypergravity was decreased up to 12 h of the exposure, however, it remained reduced under microgravity conditions, while it recovered (Act87E) and even exceeded (Act57B) the reference level under hypergravity conditions. The mRNA level of supervillin was almost unchanged. Under microgravity conditions the mRNA level of fimbrin was decreased (it recovered by the 24 h time point), while the mRNA level of alpha-actinin was significantly increased by the 12 h time point of the exposure and after 24 h it was reduced to the control level. In contrast, under hypergravity conditions the mRNA level of fimbrin initially increased, and after 24 h it dropped below the control, while the mRNA level of alpha-actinin was significantly reduced, and after 24 h it was higher than the reference level. Similar results were obtained earlier in the experiments in rodents, but similar dynamics were observed for alpha-actinin isoforms 1 and 4, although no changes were observed for fimbrin. Since Drosophila melanogaster has no alpha-actinin isoform 4, it is hypothesized that its role in the cell is played by fimbrin.
Читать
тезис
Публикация |
Влияние введения смеси фосфотидилхолинов на состояние кортикального цитоскелета волокон камбаловидной мышцы крысы
|
|
Огнева И. В. (Профессор)
Свистунов А.А. (Первый проректор)
Несвижский Юрий Владимирович (Профессор)
|
Авиакосмическая и экологическая медицина |
|
|
Цель работы - оценка состояния кортикального цитоскелета волокон камбаловидной мышцы крысы в результате 6-часового антиортостатического вывешивания на фоне предшествующего, в течение 3 сут по 100 мкг/сут, введения смеси фосфатидилхолинов (лецитина). Данные о периметре волокон, толщине подмембранного цитоскелета и доле разрывов в нем относительно периметра получали, используя иммуногистохимическую окраску на альфа-актинин-4. Периметр волокон оставался неизменным во всех группах исследования. При этом доля разрывов была выше в группах вывешивания, чем в соответствующих контрольных группах. Толщина окрашенного слоя, соответствующего подмембранному цитоскелету, не менялась в контрольных группах и в группе вывешивания без лецитина, однако в группе вывешивания на фоне лецитина она достоверно увеличивалась по сравнению с соответствующей контрольной группой на 27 % (p < 0,05) соответственно.
Purpose of the work was to assess the cortical cytoskeleton of m. soleus fibers from rats after 6-hr tail-suspension preceded by 3 days of phosphatidylcholine (lecithin) injections at a dose of 100 µг/d. Data about the fiber perimeter, submembrane cytoskeleton thickness and percentage of bonds breaks along the perimeter were obtained using the alpha-actinin-4 anti-body stain. The fiber perimeter remained unchanged in all groups under study. However, the percentage of breaks was high in suspension groups but not in respective groups of control. Thickness of the stained layer commensurate to the submembrane cytoskeleton did not change in the control groups and in the suspension group without lecithin injections but increased reliably in the suspension group that recieved injections by 27 % (p < 0.05) in comparison with its control.
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Публикация |