Heme oxygenase function is highly conserved between vertebrates where it plays important roles in normal embryonic development and controls oxidative stress. Expression of the zebrafish heme oxygenase 1 genes is known to be responsive to oxidative stress suggesting a conserved physiological function. In this study, we generate a knockout allele of zebrafish hmox1a and characterize the effects of hmox1a and hmox1b loss on embryonic development. We find that loss of hmox1a or hmox1b causes developmental defects in only a minority of embryos, in contrast to Hmox1 gene deletions in mice that cause loss of most embryos. Using a tail wound inflammation assay we find a conserved role for hmox1a, but not hmox1b, in normal macrophage migration to the wound site. Together our results indicate that zebrafish hmox1a has clearly a partitioned role from hmox1b that is more consistent with conserved functions of mammalian Heme oxygenase 1.
Get full access to this article
View all access options for this article.
References
1.
FraserST, MidwinterRG, BergerBS, StockerR. Heme oxygenase-1: a critical link between iron metabolism, erythropoiesis, and development. Adv Hematol, 2011; 2011:473709.
2.
NoguchiM, YoshidaT, KikuchiG. Specific requirement of NADPH-cytochrome c reductase for the microsomal heme oxygenase reaction yielding biliverdin IX alpha. FEBS Lett, 1979; 98:281–284.
3.
CruseI, MainesMD. Evidence suggesting that the two forms of heme oxygenase are products of different genes. J Biol Chem, 1988; 263:3348–3353.
4.
AyerA, ZarjouA, AgarwalA, StockerR. Heme oxygenases in cardiovascular health and disease. Physiol Rev, 2016; 96:1449–1508.
5.
DunnLL, MidwinterRG, NiJ, et al.New insights into intracellular locations and functions of heme oxygenase-1. Antioxid Redox Signal, 2014; 20:1723–1742.
6.
TenhunenR, MarverHS, SchmidR. Microsomal heme oxygenase. Characterization of the enzyme. J Biol Chem, 1969; 244:6388–6394.
7.
RyterSW, AlamJ, ChoiAM. Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications. Physiol Rev, 2006; 86:583–650.
8.
DwyerBE, NishimuraRN, LuSY, AlcarazA. Transient induction of heme oxygenase after cortical stab wound injury. Brain Res Mol Brain Res, 1996; 38:251–259.
9.
SunY, RotenbergMO, MainesMD. Developmental expression of heme oxygenase isozymes in rat brain. Two HO-2 mRNAs are detected. J Biol Chem, 1990; 265:8212–8217.
10.
MainesMD. The heme oxygenase system: a regulator of second messenger gases. Annu Rev Pharmacol Toxicol, 1997; 37:517–554.
11.
FujitaT, TodaK, KarimovaA, et al.Paradoxical rescue from ischemic lung injury by inhaled carbon monoxide driven by derepression of fibrinolysis. Nat Med, 2001; 7:598–604.
12.
DeveyL, FerenbachD, MohrE, et al.Tissue-resident macrophages protect the liver from ischemia reperfusion injury via a heme oxygenase-1-dependent mechanism. Mol Ther, 2009; 17:65–72.
13.
HouWH, RossiL, ShanY, et al.Iron increases HMOX1 and decreases hepatitis C viral expression in HCV-expressing cells. World J Gastroenterol, 2009; 15:4499–4510.
14.
OnoK, GotoY, TakagiS, et al.A promoter variant of the heme oxygenase-1 gene may reduce the incidence of ischemic heart disease in Japanese. Atherosclerosis, 2004; 173:315–319.
15.
YachieA, NiidaY, WadaT, et al.Oxidative stress causes enhanced endothelial cell injury in human heme oxygenase-1 deficiency. J Clin Invest, 1999; 103:129–135.
16.
OrozcoLD, KapturczakMH, BarajasB, et al.Heme oxygenase-1 expression in macrophages plays a beneficial role in atherosclerosis. Circ Res, 2007; 100:1703–1711.
17.
HolowieckiA, O'ShieldsB, JennyMJ. Characterization of heme oxygenase and biliverdin reductase gene expression in zebrafish (Danio rerio): basal expression and response to pro-oxidant exposures. Toxicol Appl Pharmacol, 2016; 311:74–87.
18.
SonnackL, KlawonnT, KriehuberR, et al.Concentration dependent transcriptome responses of zebrafish embryos after exposure to cadmium, cobalt and copper. Comp Biochem Physiol Part D Genomics Proteomics, 2017; 24:29–40.
19.
LuoK, StockerR, BrittonWJ, KikuchiK, OehlersSH. Haem oxygenase limits Mycobacterium marinum infection-induced detrimental ferrostatin-sensitive cell death in zebrafish. FEBS J, 2022. [Epub ahead of print]; DOI: 10.1111/febs.16209.
20.
ThisseC, ThisseB. High-resolution in situ hybridization to whole-mount zebrafish embryos. Nat Protoc, 2008; 3:59–69.
21.
CermakT, DoyleEL, ChristianM, et al.Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res, 2011; 39:e82.
22.
SakumaT, HosoiS, WoltjenK, et al.Efficient TALEN construction and evaluation methods for human cell and animal applications. Genes Cells, 2013; 18:315–326.
23.
SugimotoK, HuiSP, ShengDZ, NakayamaM, KikuchiK. Zebrafish FOXP3 is required for the maintenance of immune tolerance. Dev Comp Immunol, 2017; 73:156–162.
24.
WuRS, LamII, ClayH, et al.A rapid method for directed gene knockout for screening in G0 zebrafish. Dev Cell, 2018; 46:112.e4–125.e4.
25.
BlackHD, XuW, HortleE, et al.The cyclic nitroxide antioxidant 4-methoxy-TEMPO decreases mycobacterial burden in vivo through host and bacterial targets. Free Radic Biol Med, 2019; 135:157–166.
26.
ChengT, KamJY, JohansenMD, OehlersSH. High content analysis of granuloma histology and neutrophilic inflammation in adult zebrafish infected with Mycobacterium marinum. Micron, 2020; 129:102782.
27.
MattyMA, OehlersSH, TobinDM. Live imaging of host-pathogen interactions in zebrafish larvae. Methods Mol Biol, 2016; 1451:207–223.
28.
RossiA, KontarakisZ, GerriC, et al.Genetic compensation induced by deleterious mutations but not gene knockdowns. Nature, 2015; 524:230–233.
29.
KokFO, ShinM, NiCW, et al.Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish. Dev Cell, 2015; 32:97–108.
30.
KurreckJ. Antisense technologies. Improvement through novel chemical modifications. Eur J Biochem, 2003; 270:1628–1644.
31.
PrakashTP. An overview of sugar-modified oligonucleotides for antisense therapeutics. Chem Biodivers, 2011; 8:1616–1641.
32.
PossKD, TonegawaS. Heme oxygenase 1 is required for mammalian iron reutilization. Proc Natl Acad Sci U S A, 1997; 94:10919–10924.
33.
AbrahamNG. Molecular regulation—biological role of heme in hematopoiesis. Blood Rev, 1991; 5:19–28.
34.
SzadeK, ZukowskaM, SzadeA, et al.Heme oxygenase-1 deficiency triggers exhaustion of hematopoietic stem cells. EMBO Rep, 2020; 21:e47895.
RegevD, SuroliaR, KarkiS, et al.Heme oxygenase-1 promotes granuloma development and protects against dissemination of mycobacteria. Lab Invest, 2012; 92:1541–1552.
37.
WaltonEM, CronanMR, BeermanRW, TobinDM. The macrophage-specific promoter mfap4 allows live, long-term analysis of macrophage behavior during mycobacterial infection in zebrafish. PLoS One, 2015; 10:e0138949.
38.
SomloM, ClavilierL, DujonB, KermorgantM. The pho1 mutation. A frameshift, and its compensation, producing altered forms of physiologically efficient ATPase in yeast mitochondria. Eur J Biochem, 1985; 150:89–94.
39.
Carelle-CalmelsN, Saugier-VeberP, Girard-LemaireF, et al.Genetic compensation in a human genomic disorder. N Engl J Med, 2009; 360:1211–1216.
40.
El-BrolosyMA, StainierDYR. Genetic compensation: a phenomenon in search of mechanisms. PLoS Genet, 2017; 13:e1006780.
41.
FerecC, CuttingGR. Assessing the disease-liability of mutations in CFTR. Cold Spring Harb Perspect Med, 2012; 2:a009480.
42.
GenschelJ, SchmidtHH. Mutations in the LMNA gene encoding lamin A/C. Hum Mutat, 2000; 16:451–459.
43.
Silva-GomesS, AppelbergR, LarsenR, SoaresMP, GomesMS. Heme catabolism by heme oxygenase-1 confers host resistance to Mycobacterium infection. Infect Immun, 2013; 81:2536–2545.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
