| ObjectiveThis study explore the effects of CG3714gene mutation on heart rhythm under condition of exercise intervention by building drosophila arm-Ga14mutation system and anti-aging drosophila movement model established according to the characteristics of drosophila inverse gravity climbing.This study explore the effects of aging by using reverse genetics gene function research methods conditions under exercise intervention CG3714gene mutations in Drosophila changes in heart rhythm,in order to provide the reference and basis for the study of the function of anti-aging CG3714candidate genes.Metairie and methodsConstruction of arm-Gal4-CG3714mutant strain:arm-Gal4(Offered by heart development research center of Hunan Normal University) male drosophila was crossed with W1118(Offered by heart development research center of Hunan Normal University) virgin female drosophila, and the F1progeny for control group, arm-Ga14male drosophila was crossed with (w1118; P{EP} CG3714EP2611/CyO) virgin female drosophila, and the F1progeny for gene mutation group, the virgin drosophila2400was collected in each group, and the two groups were randomly divided into12groups:1,3,5-week-old arm-Gal4-W1118quiet group,1,3,5-week-old arm-Gal4-W1118exercise group,1,3,5-week-old arm-Gal4-CG3714mutant quiet group,1,3,5-week-old arm-Gal4-CG3714mutant exercise group,400/group,20/tube. Flies were kept at25℃on a12hour light/12hour dark cycle and50%humidity.Using drosophila sports training platform (made by Hunan Normal University fitness and sports rehabilitation key laboratory) for Training, Expression of CG3714mRNA was measured by real-time quantitative PCR assay in12hours after the exercise, and Flies heart rate, systolic diameter, systolic interval, diastolic interval, heart period, arrhythmia index and fibrillation were detected by M-mode echocardiography. Climbing index calculation and statistical life span.Results3.1The results of mRNA in DrosophilaThe result of mRNA show that, the average relative expression(RQ) content of CG3714mRNA in the mutation group was less1/20times than the respective arm-Gal4-W1118control group, indicating that CG3714drosophila mutation makes mRNA expression level was reduced obviously, basically did not express the gene CG3714protein proved that the gene mutation was set up successfully.3.2The results of heart M-Mode in drosophila3.2.1The results of hear rate in drosophila Drosophilas’heart rate increased with age and decreased gradually; arm-Gal4-W1118strains of quiet group Drosophilas peers heart rate greater than their in the exercise group, and the heart rate have a great difference between the exercise group and the quiet in one week old Drosophilas (P<0.01), indicating the exercise can reduce the Drosophilas’heart rate effectively. The peers heart rate in the arm-Gal4-CG3714gene mutation exercise group was greater than that in the arm-Gal4-CG3714mutant quiet group, indicating that CG3714gene mutation Drosophila can lead to increased heart rate.3.2.2The results of systole period, diastolic period and cardiac cycle in drosophilaDrosophilas’cardiac systole period, diastolic period and cardiac cycle are showing an increasing trend with age increasing; exercise can increase Drosophilas’systolic period effectively, both diastolic period and cardiac cycle are increasing heavily; CG3714mutation in Drosophila heart has a little of effect on diastolic period, but it can increased inhibition of both the drosophilas’diastolic period and cardiac cycle.3.2.3The result of drosophilas’arrhythmia indexDrosophila arrhythmia index increases with age showing a rising trend, exercise can make Drosophila cardiac arrhythmias rose, CG3714mutations in drosophila cardiac arrhythmia index rose, implemented on the basis of gene mutations CG3714motion to intervene so that the magnitude of arrhythmia index rose increases.3.2.4The result of drosophilas’ cardiac fibrosis tremorDrosophilas’ cardiac fibrous tremor occurred with increasing age showing a rising trend, the greater the age, the more severer tremor fiber; movement increases the fiber tremor occurred; CG3714Drosophila gene Exercise promotes cardiac fibers tremor occurred, based on the implementation of the CG3714mutation occur in Drosophila cardiac motion to intervene so that the fibers tremor worsened, it is not conducive to a stable Drosophila heart rhythm.3.3The result of drosophilas’ life spanarm-Gal4-W1118exercise group flies lifetime span is higher than half of the deaths lifetime value arm-Gal4-W1118control flies(P<0.05); the average life expectancy arm-Gal4-CG3714mutant flies exercise group was lower than the arm-Gal4-CG3714Drosophila lifespan in the control group at the same age (P<0.05); The maximum life span of fruit flies in the detection of the arm-Gal4-CG3714mutant flies quiet group lower than arm-Gal4-W1118quiet group at the same age (P<0.05); arm-Ga14-CG3714mutant exercise group highest lifetime value lower than the arm-Gal4-CG3714mutant quiet group at the same age(P<0.05).Compared with the control group, arm-Gal4-W1118exercise group’s average life extension rate was increased, arm-Gal4-CG3714control group’s life extension rate was decreased compared with the arm-Gal4-W1118control group, while the arm-Gal4-CG3714drosophilas’ average life extension rate decreased significantly under the condition of the exercise intervention.3.4The result of drosophilas’climbing abilityThe ability to climb marked decline in drosophila with aging, while arm-Gal4-W1118exercise group’s climbing ability improved obviously under the condition of the exercise training, indicating that exercise training delays the drosophilas’climbing ability to decline. The arm-Ga14-CG3714gene mutation decreased drosophilas’climbing capacity and exacerbated the decline in fruit flies climbing ability with exercise intervention.4. Conclusion4.1The regular exercise intervene can effectively decrease fruit flies heart rate, increase fruit flies systole period and diastolic period and the cardiac cycle, prompted flies prolong life span delays the drosophilas’climbing ability to decline; but the exercise will lead to cardiac arrhythmia index rose and fibrous tremor.4.2CG3714mutations made CG3714mRNA expression decreased significantly, made fruit flies the heart rate increases, the flies between cardiac cycle and heart diastolic phase of the increase of inhibition, prompting flies the occurrence of cardiac arrhythmia index rose and fibrous tremor, result in drosophila life cycle shorten, athletic ability decline.4.3The exercise interference obvious increase in heart rate, make the fruit flies arrhythmia index rise obviously, can make flies tremor fibers aggravated and cause drosophila arrhythmia, resulting in drosophilas heart rhythm disorders based on CG3714gene mutation, the heart rhythm is not conducive to stable flies. We can speculate that the gene CG3714has the function to reduce the incidence of arrhythmias and fiber tremor caused by aging, and make a smooth rhythm in drosophila, also to protect normal cardiac function; The gene CG3714can improve exercise capacity and extend the life of drosophila and other functions. |
