Abstract
Our study was aimed to investigate the effects of lgals3a (Gal-3 encoding gene) on the development of zebrafish embryo and its underlying mechanisms. Morpholino (MO) technology was used to inhibit the expression of zebrafish lgals3a, and the effect of lgals3a gene knockdown on zebrafish embryo development and the number of monocyte macrophages was observed. Effect of lgals3a-e3i3-MO on apoptosis of zebrafish was detected by acridine orange staining. In addition, the mRNA expression levels of Wnt/β-catenin signaling pathway-related genes were detected by RT-qPCR. Compared with control-MO group, the zebrafish embryos injected with lgals3a-e3i3-MO had obvious defects in the head, eyes, and tail, and pericardial edema. Lgals3a-e3i3-MO significantly reduced the number of mononuclear macrophages in zebrafish embryos compared with the control-MO group. The results of acridine orange staining showed that compared with the control-MO group, lgals3a-e3i3-MO promoted cardiomyocyte apoptosis in zebrafish. Furthermore, lgals3a-e3i3-MO significantly up-regulated the expression of dkk1b, wnt9a, lrp5, fzd7a, β-catenin, Gsk-3β, mycn, myca in the Wnt/β-catenin pathway, and decreased the expression of lef1. These results indicate that lgals3a-e3i3-MO inhibits zebrafish embryo development, reduces the number of mononuclear macrophages, activates Wnt/β-catenin signaling pathway and promotes cardiomyocyte apoptosis.
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References
Ahmed H, Du SJ, O’Leary N, Vasta GR (2004) Biochemical and molecular characterization of galectins from zebrafish (Danio rerio): notochord-specific expression of a prototype galectin during early embryogenesis. Glycobiology 14:219–232. https://doi.org/10.1093/glycob/cwh032
Baptiste TA, James A, Saria M, Ochieng J (2007) Mechano-transduction mediated secretion and uptake of galectin-3 in breast carcinoma cells: implications in the extracellular functions of the lectin. Exp Cell Res 313:652–664. https://doi.org/10.1016/j.yexcr.2006.11.005
Basu S, Sachidanandan C (2013) Zebrafish: a multifaceted tool for chemical biologists. Chem Rev 113:7952–7980. https://doi.org/10.1021/cr4000013
Blanchard H, Yu X, Collins PM, Bum-Erdene K (2014) Galectin-3 inhibitors: a patent review (2008–present). Expert Opin Ther Pat 24:1053–1065. https://doi.org/10.1517/13543776.2014.947961
Clevers H (2006) Wnt/ beta-catenin signaling in development and disease. Cell 127:469–480. https://doi.org/10.1016/j.cell.2006.10.018
Colnot C, Ripoche M, Milon G, Montagutelli X, Poirier F (2010) Maintenance of granulocyte numbers during acute peritonitis is defective in galectinnull mutant mice. Immunology 94:290–296. https://doi.org/10.1046/j.1365-2567.1998.00517.x
De Boer RA, van Veldhuisen DJ, Gansevoort RT, Muller Kobold AC, van Gilst WH, Hillege HL, Bakker SJL, van der Harst P (2012) The fibrosis marker galectin-3 and outcome in the general population. J Intern Med 272:55–64. https://doi.org/10.1111/j.1365-2796.2011.02476.x
Ebrahim AH, Alalawi Z, Mirandola L, Rakhshanda R, Dahlbeck S, Nguyen D, Jenkins M, Grizzi F, Cobos E, Figueroa JA, Chiriva-Internati M (2014) Galectins in cancer: carcinogenesis, diagnosis and therapy. Ann Transl Med 2:88. https://doi.org/10.3978/j.issn.2305-5839.2014.09.12
Fairbairn EA, Bonthius J, Cherr GN (2012) Polycyclic aromatic hydrocarbons and dibutyl phthalate disrupt dorsal-ventral axis determination via the Wnt/β-catenin signaling pathway in zebrafish embryos. Aquat Toxicol 124–125:188–196. https://doi.org/10.1016/j.aquatox.2012.08.017
Fortuna-Costa A, Gomes AM, Kozlowski EO, Stelling MP, Pavão MS (2014) Extracellular galectin-3 in tumor progression and metastasis. Front Oncol 4:138. https://doi.org/10.3389/fonc.2014.00138
Glinskii OV, Sud S, Mossine VV, Mawhinney TP, Anthony DC, Glinsky GV, Pienta KJ, Glinsky VV (2012) Inhibition of prostate cancer bone metastasis by synthetic TF antigen mimic/galectin-3 inhibitor lactulose-L-leucine. Neoplasia 14:65–73. https://doi.org/10.1593/neo.111544
Henderson NC, Sethi T (2009) The regulation of inflammation by galectin-3. Immunol Rev 230:160–171. https://doi.org/10.1111/j.1600-065X.2009.00794.x
Ikeda S, Kishida S, Yamamoto H, Murai H, Koyama S, Kikuchi A (1998) Axin, a negative regulator of the Wnt signaling pathway, forms a complex with GSK-3β and β-catenin and promotes GSK-3β-dependent phosphorylation of β-catenin. EMBO J 17:1371–1384. https://doi.org/10.1093/emboj/17.5.1371
Klyosov AA, Traber PG (2012) Galectins in disease and potential therapeutic approaches. In: Galectins and disease implications for targeted therapeutics. ACS Publications, pp 3–43. https://doi.org/10.1021/bk-2012-1115.ch001
MacKinnon AC, Liu X, Hadoke PW, Miller MR, Newby DE, Sethi T (2013) Inhibition of galectin-3 reduces atherosclerosis in apolipoprotein E-deficient mice. Glycobiology 23:654–663. https://doi.org/10.1093/glycob/cwt006
Nachtigal M, Ghaffar A, Mayer EP (2008) Galectin-3 Gene inactivation reduces atherosclerotic lesions and adventitial inflammation in ApoE-deficient mice. Am J Pathol 172:247–255. https://doi.org/10.2353/ajpath.2008.070348
Nasevicius A, Ekker SC (2000) Effective targeted gene ‘knockdown’ in zebrafish. Nat Genet 26:216–220. https://doi.org/10.1038/79951
Nelson WJ, Nusse R (2004) Convergence of Wnt, ß-catenin, and cadherin pathways. Science 303:1483–1487. https://doi.org/10.1126/science.1094291
Newlaczyl AU, Yu L-G (2011) Galectin-3–a jack-of-all-trades in cancer. Cancer Lett 313:123–128. https://doi.org/10.1016/j.canlet.2011.09.003
Papaspyridonos M, McNeill E, de Bono JP, Smith A, Burnand KG, Channon KM, Greaves DR (2008) Galectin-3 is an amplifier of inflammation in atherosclerotic plaque progression through macrophage activation and monocyte chemoattraction. Arterioscler Thromb Vasc Biol 28:433–440. https://doi.org/10.1161/ATVBAHA.107.159160
Pereira AR, Falcão LM (2015) Galectin-3, a prognostic marker–and a therapeutic target? Rev Port Cardiol 34:201–208. https://doi.org/10.1016/j.repc.2014.10.005
Santoriello C, Zon LI (2012) Hooked! modeling human disease in zebrafish. J Clin Invest 122:2337–2343. https://doi.org/10.1172/JCI60434
Shu W, Guttentag S, Wang Z, Andl T, Ballard P, Lu MM, Piccolo S, Birchmeier W, Whitsett JA, Millar SE, Morrisey EE (2005) Wnt/β-catenin signaling acts upstream of N-myc, BMP4, and FGF signaling to regulate proximal–distal patterning in the lung. Dev Biol 283:226–239. https://doi.org/10.1016/j.ydbio.2005.04.014
Smirnova A, Mentor A, Ranefall P, Bornehag CG, Brunström B, Mattsson A, Jönsson M (2021) Increased apoptosis, reduced Wnt/β-catenin signaling, and altered tail development in zebrafish embryos exposed to a human-relevant chemical mixture. Chemosphere 264:128467. https://doi.org/10.1016/j.chemosphere.2020.128467
Song L, Tang JW, Owusu L, Sun M-Z, Wu J, Zhang J (2014) Galectin-3 in cancer. Clin Chim Acta 431:185–191. https://doi.org/10.1016/j.cca.2014.01.019
Ueno S, Weidinger G, Osugi T, Kohn AD, Golob JL, Pabon L, Reinecke H, Moon RT, Murry CE (2007) Biphasic role for Wnt/β-catenin signaling in cardiac specification in zebrafish and embryonic stem cells. Proc Natl Acad Sci U S A 104:9685–9690. https://doi.org/10.1073/pnas.0702859104
Üstündağ ÜV, Ünal İ, Ateş PS, Alturfan AA, Yiğitbaşı T, Emekli-Alturfan E (2017) Bisphenol A and di(2-ethylhexyl) phthalate exert divergent effects on apoptosis and the Wnt/β-catenin pathway in zebrafish embryos: a possible mechanism of endocrine disrupting chemical action. Toxicol Ind Health 33:901–910. https://doi.org/10.1177/0748233717733598
Wallace KN, Pack M (2003) Unique and conserved aspects of gut development in zebrafish. Dev Biol 255:12–29. https://doi.org/10.1016/s0012-1606(02)00034-9
Yu X, Sun Y, Zhao Y, Zhang W, Yang Z, Gao Y, Cai H, Li Y, Wang Q, Bian B, Nie J (2015) Prognostic value of plasma galectin-3 levels in patients with coronary heart disease and chronic heart failure. Int Heart J 56:314–318. https://doi.org/10.1536/ihj.14-304
Zhang H, Luo M, Liang X, Wang D, Gu X, Duan C, Gu H, Chen G, Zhao X, Zhao Z, Liu C (2014) Galectin-3 as a marker and potential therapeutic target in breast cancer. PLoS ONE 9:e103482. https://doi.org/10.1371/journal.pone.0103482
Zhang Y, Bai XT, Zhu KY, Jin Y, Deng M, Le HY, Fu YF, Chen Y, Zhu J, Look AT, Kanki J, Chen Z, Chen SJ, Liu TX (2008) In vivo interstitial migration of primitive macrophages mediated by JNK-matrix metalloproteinase 13 signaling in response to acute injury. J Immunol 181:2155–2164. https://doi.org/10.4049/jimmunol.181.3.2155
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This work was supported by the National Natural Science Foundation of China [grant number 81500331] and the Science and Technology Commission of Shanghai [grant number 17140902600].
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CW conceived and designed the study. KC, CW, YF, JG, ZH, YW, HZ, CM and LG performed the main experiments and analyzed the data. KC and CW wrote the manuscript. All the authors have read and approved the final version of the manuscript.
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Chen, K., Fan, Y., Gu, J. et al. Effect of lgals3a on embryo development of zebrafish. Transgenic Res 30, 739–750 (2021). https://doi.org/10.1007/s11248-021-00276-5
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DOI: https://doi.org/10.1007/s11248-021-00276-5