MiR92b-3p Synthetic Analogue Impairs Zebrafish Embryonic Development, Leading to Ocular Defects, Decreased Movement and Hatching Rate, and Increased Mortality

Overview

Journal J Appl Genet

Publisher Springer

Specialty Genetics

Date 2022 Oct 23

PMID 36274083

Authors

Kilian Kranert

Maciej Wozny

Piotr Podlasz

Krzysztof Wasowicz

Pawel Brzuzan

Affiliations

Soon will be listed here.

Abstract

The aim of this study was to examine the effect of microRNA 92b-3p (MiR92b-3p) overexpression on the embryonic development of zebrafish. A synthetic MiR92b-3p analogue (mirVana™ mimic, in vivo-ready) was injected at doses up to 5 ng/embryo into the yolk sac of embryos (2-16 cell stage). At 24 h post fertilization (hpf), the locomotor activity of the embryos was measured, and after hatching (72 hpf), the rates of malformation occurrence, hatching, and mortality were determined. Next, the larvae were fixed for histological and molecular examinations. Exposure to the MiR92b-3p mimic impaired embryonic development, leading to increased occurrence of malformations (i.e., pericardial edema, spine curvature, smaller eyes), decreased locomotor activity and hatching rate, and increased mortality. Importantly, the mimic affected retinal differentiation and lens formation during zebrafish embryogenesis, which suggests that MiR92b-3p could be an important factor in the regulation of fish embryogenesis and ocular development. The expression level of MiR92b-3p was substantially higher in the exposed larvae than in the untreated larvae, indicating that the mimic was successfully delivered to the zebrafish. Although screening of potential MiR92b-3p target genes suggested some changes in their expression levels, these results were inconclusive. Together, this study indicates that MiR92b-3p mimic impairs zebrafish embryonic development, and further research is necessary to identify the MiR92b-3p-regulated cell pathways involved in the impairment of the fish's development.

References

Kleinjan D, Bancewicz R, Gautier P, Dahm R, Schonthaler H, Damante G . Subfunctionalization of duplicated zebrafish pax6 genes by cis-regulatory divergence. PLoS Genet. 2008; 4(2):e29. PMC: 2242813. DOI: 10.1371/journal.pgen.0040029. View

Amrhein V, Korner-Nievergelt F, Roth T . The earth is flat ( > 0.05): significance thresholds and the crisis of unreplicable research. PeerJ. 2017; 5:e3544. PMC: 5502092. DOI: 10.7717/peerj.3544. View

Wang G, Cheng B, Jia R, Tan B, Liu W . Altered expression of microRNA-92b-3p predicts survival outcomes of patients with prostate cancer and functions as an oncogene in tumor progression. Oncol Lett. 2020; 21(1):4. PMC: 7681231. DOI: 10.3892/ol.2020.12265. View

Kloosterman W, Steiner F, Berezikov E, de Bruijn E, van de Belt J, Verheul M . Cloning and expression of new microRNAs from zebrafish. Nucleic Acids Res. 2006; 34(9):2558-69. PMC: 3303176. DOI: 10.1093/nar/gkl278. View

Basnet R, Guarienti M, Memo M . Zebrafish Embryo as an In Vivo Model for Behavioral and Pharmacological Characterization of Methylxanthine Drugs. Int J Mol Sci. 2017; 18(3). PMC: 5372612. DOI: 10.3390/ijms18030596. View

Leach L, Hanovice N, George S, Gabriel A, Gross J . The immune response is a critical regulator of zebrafish retinal pigment epithelium regeneration. Proc Natl Acad Sci U S A. 2021; 118(21). PMC: 8166181. DOI: 10.1073/pnas.2017198118. View

Reiter J, Kikuchi Y, Stainier D . Multiple roles for Gata5 in zebrafish endoderm formation. Development. 2000; 128(1):125-35. DOI: 10.1242/dev.128.1.125. View

Deveale B, Swindlehurst-Chan J, Blelloch R . The roles of microRNAs in mouse development. Nat Rev Genet. 2021; 22(5):307-323. DOI: 10.1038/s41576-020-00309-5. View

Cumming G . The new statistics: why and how. Psychol Sci. 2013; 25(1):7-29. DOI: 10.1177/0956797613504966. View

10.

Majewski M, Kasica N, Jakimiuk A, Podlasz P . Toxicity and cardiac effects of acute exposure to tryptophan metabolites on the kynurenine pathway in early developing zebrafish (Danio rerio) embryos. Toxicol Appl Pharmacol. 2018; 341:16-29. DOI: 10.1016/j.taap.2018.01.004. View

11.

Jonas S, Izaurralde E . Towards a molecular understanding of microRNA-mediated gene silencing. Nat Rev Genet. 2015; 16(7):421-33. DOI: 10.1038/nrg3965. View

12.

Macaulay I, Svensson V, Labalette C, Ferreira L, Hamey F, Voet T . Single-Cell RNA-Sequencing Reveals a Continuous Spectrum of Differentiation in Hematopoietic Cells. Cell Rep. 2016; 14(4):966-977. PMC: 4742565. DOI: 10.1016/j.celrep.2015.12.082. View

13.

Wienholds E, Kloosterman W, Miska E, Alvarez-Saavedra E, Berezikov E, de Bruijn E . MicroRNA expression in zebrafish embryonic development. Science. 2005; 309(5732):310-1. DOI: 10.1126/science.1114519. View

14.

Takamiya M, Weger B, Schindler S, Beil T, Yang L, Armant O . Molecular description of eye defects in the zebrafish Pax6b mutant, sunrise, reveals a Pax6b-dependent genetic network in the developing anterior chamber. PLoS One. 2015; 10(2):e0117645. PMC: 4334901. DOI: 10.1371/journal.pone.0117645. View

15.

Kozomara A, Birgaoanu M, Griffiths-Jones S . miRBase: from microRNA sequences to function. Nucleic Acids Res. 2018; 47(D1):D155-D162. PMC: 6323917. DOI: 10.1093/nar/gky1141. View

16.

Cheng R, Jia Y, Dai L, Liu C, Wang J, Li G . Tris(1,3-dichloro-2-propyl) phosphate disrupts axonal growth, cholinergic system and motor behavior in early life zebrafish. Aquat Toxicol. 2017; 192:7-15. DOI: 10.1016/j.aquatox.2017.09.003. View

17.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7):RESEARCH0034. PMC: 126239. DOI: 10.1186/gb-2002-3-7-research0034. View

18.

Long M, Zhan M, Xu S, Yang R, Chen W, Zhang S . miR-92b-3p acts as a tumor suppressor by targeting Gabra3 in pancreatic cancer. Mol Cancer. 2017; 16(1):167. PMC: 5659029. DOI: 10.1186/s12943-017-0723-7. View

19.

Li N, Wei C, Olena A, Patton J . Regulation of endoderm formation and left-right asymmetry by miR-92 during early zebrafish development. Development. 2011; 138(9):1817-26. PMC: 3074454. DOI: 10.1242/dev.056697. View

20.

Wang X . Composition of seed sequence is a major determinant of microRNA targeting patterns. Bioinformatics. 2014; 30(10):1377-83. PMC: 4016705. DOI: 10.1093/bioinformatics/btu045. View