Analysis of heart function in flies expressing unc-45dsRNA.Scer\UAS.cUa under the control of Scer\GAL4HCH.Hand reveals severe heart defects compared with controls. Hearts with cardiac unc-45dsRNA.Scer\UAS.cUa-expression are significantly dilated in both the conical chamber and the third abdominal segment heart and exhibit dramatically arrhythmic beating patterns, compared with aged-matched control hearts. Severe cardiac dysfunction is observed as early as late pupal stages. In contrast, no major apparent cardiac defects are observed in hearts of third instar larvae and young (white) pupae expressing unc-45dsRNA.Scer\UAS.cUa via Scer\GAL4HCH.Hand regarding cardiac contractility compared to wild-type. However, quantitative analyses of cardiac data obtained from both third instar larvae and young pupae reveal that the unc-45dsRNA.Scer\UAS.cUa-expressing hearts show a small but significant increase in heart rate (tachycardia) compared with age-matched control. In addition to tachycardia, expression of unc-45dsRNA.Scer\UAS.cUa in third instar larval hears results in mild but significant cardiac dilation, which is more evident after metamorphosis. Both diastolic and systolic diameters of unc-45dsRNA.Scer\UAS.cUa-expressing third instar larval hearts are significantly larger than control hearts. Although unc-45dsRNA.Scer\UAS.cUa-expressing third instar larval hearts show tachycardia, their cardiac performance is depressed compared with control third instar larvae.
Conditional expression of unc-45dsRNA.Scer\UAS.cUa under the control of both Scer\GAL4tin.CΔ4 and Scer\GAL80ts.αTub84B results in a severe cardiac phenotype in adult hearts throughout life. For example, one week old hearts conditionally expressing unc-45dsRNA.Scer\UAS.cUa beginning before metamorphosis (third instar larval stage) show prolonged systolic and diastolic intervals compared with controls. In addition, such unc-45dsRNA.Scer\UAS.cUa-expressing hearts are dilated, their fractional shortening is reduced, and cardiac arrhythmias are increased. Severe cardiac defects are also apparent in newly enclosed adults and late pupae when unc-45dsRNA.Scer\UAS.cUa expression is induced at the beginning of the pupal stage instead of the third instar larval stage. In contrast to the above date, when unc-45dsRNA.Scer\UAS.cUa is expressed in the heart after metamorphosis (adult flies), dilation of the heart does not occur, however, systolic and diastolic intervals are prolonged and arrhythmias are elevated. Similar results are obtained when examining the hearts of older flies.
Quantitative analysis of various cardiac physiological parameters in 1-3 week old fly hearts expressing unc-45dsRNA.Scer\UAS.cUa via Scer\GAL4HCH.Hand reveals a significant prolongation of the heartbeat length, compared with wild-type, that manifests in an increase of both systolic and diastolic intervals. Particularly dramatic is the cardiac dilation due to both systolic and diastolic heart diameter increases in unc-45dsRNA.Scer\UAS.cUa-expressing hearts compared with control hearts. The observed extreme cardiac dilation is accompanied by a significant reduction in heart contractility, which is quantified in a decreased fractional shortening. The incidence of arrhythmias is also much higher in unc-45dsRNA.Scer\UAS.cUa-expressing hearts than that in controls. This is in large part due to the frequent occurrence of asystolies, intermittent stoppages in heartbeat.
Cardiac expression of unc-45dsRNA.Scer\UAS.cUa under the control of Scer\GAL4HCH.Hand has a drastic negative impact on the life-span of flies.
Mesodermal expression of unc-45dsRNA.Scer\UAS.cUa under the control of Scer\GAL4how-24B results in early developmental lethality.
Myofibrillar disorganization is apparent in hearts expressing unc-45dsRNA.Scer\UAS.cUa under the control of Scer\GAL4HCH.Hand compared with wild-type hearts.
unc-45+t significantly improves the cardiac phenotypes and life-span defects resulting from the expression of unc-45dsRNA.Scer\UAS.cUa under the control of Scer\GAL4HCH.Hand.