Fig_5.tif (1.49 MB)

DGAT-2 and ACS-22 exhibit redundant and diet-dependent effects on the longevity and lipid content of germline-less mutants.

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posted on 2016-02-10, 10:39 authored by Francis Raj Gandhi Amrit, Elizabeth Marie Steenkiste, Ramesh Ratnappan, Shaw-Wen Chen, T. Brooke McClendon, Dennis Kostka, Judith Yanowitz, Carissa Perez Olsen, Arjumand Ghazi

A. RNAi inactivation ‘dgat’ genes shortens glp-1 mutants longevity modestly. glp-1 mutants were subjected to RNAi during adulthood by feeding bacteria containing empty control vector (green) or bacteria expressing dsRNA targeting ‘dgat’ genes. Control (green; m = 23.2 ± 0.3, n = 63/83), daf-16 (red; m = 15.2 ± 0.3, n = 79/79; P vs. control <0.0001), dgat-2 (pink; m = 18.5 ± 0.7, n = 71/80; P vs. control <0.0001), acs-22 (purple, m = 19.7 ± 1.3, n = 123/123, P vs. control, 0.006), Y53G8B.2 (blue, m = 20.7 ± 0.5, n = 59/71, P vs. control 0.02) and K07B1.4 (sand, m = 19.9 ± 1.0, n = 97/97, P vs. control, <0.0001). B, C. Effect of dgat-2(hj44) and acs-22(hj26) mutations on the lifespan of glp-1 mutants and wild-type (N2) worms on E. coli OP50. B.glp-1 (green; m = 26.5 ± 0.5, n = 76/85), dgat-2;glp-1 (pink; m = 28.5 ± 1.0, n = 52/61; P vs. glp-1 0.64), N2 (black; m = 16.7 ± 0.7, n = 76/91, P vs. glp-1 <0.0001), dgat-2 (gray; m = 22.6 ± 0.6, n = 75/88; P vs. N2 <0.0001). C:glp-1 (green; m = 21.0 ± 0.9, n = 92/96), acs-22;glp-1 (purple; m = 23.6 ± 0.6, n = 97/103; P vs. glp-1 0.87), N2 (black; m = 17.0 ± 0.7, n = 69/100), acs-22 (gray; m = 19.1 ± 0.5, n = 72/83; P vs. N2 0.1). Additional trials are shown in S6B and S6C and S7(A) Tables. D. Effect of dgat-2(hj44) and acs-22(hj26) mutations on the lifespan of glp-1 mutants grown during adulthood on E. coli HT115 with the empty control vector. N2 (black; m = 19.16 ± 0.5, n = 57/75), glp-1 (green; m = 24.2 ± 1.6, n = 43/60), dgat-2;glp-1 (pink; m = 16.3 ± 0.4, n = 87/98; P vs. glp-1 <0.0001), acs-22;glp-1 (purple; m = 15.7 ± 0.4, n = 75/86). E, F. Effect of dgat-2(hj44), acs-22(hj26) and fasn-1(fr8) mutations on fat levels of glp-1 mutants. Quantification of lipid levels compared between different strains through ORO staining on day 2 adults grown on E. coli OP50 (E) or E. coli HT115 (F). The mutations caused reduced ORO staining in both fertile and glp-1 backgrounds when grown on E. coli OP50. On E. coli HT115, dgat-2;glp-1 and acs-22;glp-1 did not show a significant ORO diminution. The box plots illustrate data from two biological replicates tested for each strain. Asterisks represent the P values {<0.05 (*), <0.005 (**) or <0.0001 (***)} derived using the multcomp package. See S6 Fig for ORO data of day 4 and 8 adults, and Methods for details of the statistical analyses.