Experimental selection of hypoxia-tolerant Drosophila melanogaster.
Through long-term laboratory selection (over 200 generations), we have generated Drosophila melanogaster populations that tolerate severe, normally lethal, levels of hypoxia. Because of initial experiments suspecting genetic mechanisms underlying this adaptation, we compared the genomes of the hypoxia-selected flies with those of controls using deep resequencing. By applying unique computing and analytical methods we identified a number of DNA regions under selection, mostly on the X chromosome. Several of the hypoxia-selected regions contained genes encoding or regulating the Notch pathway. In addition, previous expression profiling revealed an activation of the Notch pathway in the hypoxia-selected flies. We confirmed the contribution of Notch activation to hypoxia tolerance using a specific γ-secretase inhibitor, N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT), which significantly reduced adult survival and life span in the hypoxia-selected flies. We also demonstrated that flies with loss-of-function Notch mutations or RNAi-mediated Notch knockdown had a significant reduction in hypoxia tolerance, but those with a gain-of-function had a dramatic opposite effect. Using the UAS-Gal4 system, we also showed that specific overexpression of the Notch intracellular domain in glial cells was critical for conferring hypoxia tolerance. Unique analytical tools and genetic and bioinformatic strategies allowed us to discover that Notch activation plays a major role in this hypoxia tolerance in Drosophila melanogaster.
Department of Pediatrics, University of California at San Diego, La Jolla, CA 92093, USA.
Dan
D
Zhou
Nitin
N
Udpa
Merril
M
Gersten
DeeAnn W
DW
Visk
Ali
A
Bashir
Jin
J
Xue
Kelly A
KA
Frazer
James W
JW
Posakony
Shankar
S
Subramaniam
Vineet
V
Bafna
Gabriel G
GG
Haddad
D000222:Adaptation, Physiological / Q000187:drug effects / Q000235:genetics*; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000818:Animals; D059006:Chromosomes, Insect / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004151:Dipeptides / Q000494:pharmacology; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster; D000860:Hypoxia / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D017434:Protein Structure, Tertiary; D051880:Receptors, Notch / Q000235:genetics* / Q000378:metabolism; D012641:Selection, Genetic; D014960:X Chromosome / Q000235:genetics* / Q000378:metabolism
D004151:Dipeptides; D029721:Drosophila Proteins; C497759:N protein, Drosophila; C419410:N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester; D051880:Receptors, Notch; D004247:DNA; D053829:Amyloid Precursor Protein Secretases
Nature. 1986 Sep 4-10;323(6083):89-91
3018588
Mol Cell Biol. 2005 Dec;25(23):10433-41
16287856
J Hered. 2009 Jan-Feb;100(1):97-105
18836147
PLoS Genet. 2008 Oct;4(10):e1000221
18927626
Dev Cell. 2005 Nov;9(5):617-28
16256737
Lab Invest. 2008 Jan;88(1):11-7
18059366
Development. 1993 Jun;118(2):401-15
8223268
Nature. 1988 Apr 28;332(6167):853-6
3128741
Nucleic Acids Res. 2000 Jan 1;28(1):27-30
10592173
Genetics. 1975 Sep;81(1):143-62
812766
Proc Natl Acad Sci U S A. 2008 Apr 29;105(17):6392-7
18427106
Trends Cardiovasc Med. 2007 Jul;17(5):177-82
17574126
Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3368-73
18299578
Nature. 2010 Sep 30;467(7315):587-90
20844486
Proc Natl Acad Sci U S A. 2010 Apr 13;107(15):6882-7
20339081
Science. 2010 Jul 2;329(5987):75-8
20595611
Nature. 1991 Jun 20;351(6328):652-4
1904993
Proc Natl Acad Sci U S A. 2002 May 14;99(10):7021-6
12011461
J Clin Invest. 2008 Nov;118(11):3660-70
18924608
EMBO J. 1987 Nov;6(11):3407-17
3123218
Nat Immunol. 2005 Sep;6(9):881-8
16056227
Oncogene. 2010 Apr 29;29(17):2488-98
20154720
PLoS One. 2007 May 30;2(5):e490
17534440
Physiol Genomics. 2005 Aug 11;22(3):292-307
15928208
Mech Dev. 1999 Aug;86(1-2):39-50
10446264
Nat Rev Genet. 2005 Jan;6(1):9-23
15630418
Cell Metab. 2007 Oct;6(4):257-66
17908555
FASEB J. 2006 Jun;20(8):1275-7
16645044
J Neurochem. 2001 Jan;76(1):173-81
11145990
Genome Res. 2001 May;11(5):863-74
11337480
Genome Res. 2008 Nov;18(11):1851-8
18714091
Nature. 2006 Aug 17;442(7104):823-6
16799564
J Mol Evol. 2005 Sep;61(3):315-24
16044248
Nat Med. 2009 Nov;15(11):1289-97
19855400
Genetics. 1989 Nov;123(3):585-95
2513255
Nat Rev Immunol. 2007 Jan;7(1):64-75
17170755
Genetics. 1993 Mar;133(3):693-709
8454210
Nat Rev Mol Cell Biol. 2006 Sep;7(9):678-89
16921404
Annu Rev Pathol. 2009;4:315-42
18842101
Stem Cells. 2010 Nov;28(11):1918-29
20827750