Development of tg(Uas:Sec‐hsa.anxa5‐yfp,myl7:Rfp); casper(roy−/−,nacre−/−) transparent transgenic in vivo zebrafish model to study the cardiomyocyte function

Surendra K. Rajpurohit, Aaron Gopal, May Ye Mon, Nikhil G. Patel, Vishal Arora

Research output: Contribution to journalArticlepeer-review

Abstract

The zebrafish provided an excellent platform to study the genetic and molecular approach of cellular phenotype‐based cardiac research. We designed a novel protocol to develop the transparent transgenic zebrafish model to study annexin‐5 activity in the cardiovascular function by gen-erating homozygous transparent skin Casper (roy−/−,nacre−/−); myl7:RFP; annexin‐5:YFP transgenic zebrafish. The skin pigmentation background of any vertebrate model organism is a major obstruc-tion for in vivo confocal imaging to study the transgenic cellular phenotype‐based study. By developing Casper (roy−/−,nacre−/− ); myl7; annexin‐5 transparent transgenic zebrafish strain, we estab-lished time‐lapse in vivo confocal microscopy to study cellular phenotype/pathologies of cardiomy-ocytes over time to quantify changes in cardiomyocyte morphology and function over time, com-paring control and cardiac injury and cardio‐oncology. Casper contributes to the study by integrat-ing a transparent characteristic in adult zebrafish that allows for simpler transparent visualization and observation. The Casper (roy−/−,nacre−/− ) transgenic progenies developed through cross‐breed-ing with the transgenic strain of Tg (UAS:SEC‐Hsa.ANXA5‐YFP,myl7:RFP). Confocal and fluorescent microscopy were being used to obtain accurate, precise imaging and to determine fluorescent protein being activated. This study protocol was conducted under two sections; 1.1: Generation of homozygous Tg (UAS:SEC‐Hsa.ANXA5‐YFP,myl7:RFP); Casper (roy−/−,nacre−/− ) zebrafish (generation F01‐F06) and 1.2: Screening and sorting the transparent transgenic progeny and in vivo imaging to validate cardiac morphology through in vivo confocal imaging. We coined the newly developed strain as Tg (UAS:SEC‐Hsa.ANXA5‐YFP,myl7:RFP); Casper (roy−/−,nacre−/−)gmc1. Thus, the newly developed strain maintains transparency of the skin throughout the entire life of zebrafish and is ca-pable of application of a non‐invasive in vivo imaging process. These novel results provide an in vivo whole organism‐based platform to design high‐throughput screening and establish a new hori-zon for drug discovery in cardiac cell death and cardio‐oncology therapeutics and treatment.

Original languageEnglish (US)
Article number1963
JournalCells
Volume10
Issue number8
DOIs
StatePublished - Aug 2021

Keywords

  • Annexin‐5
  • Cardiomyocyte
  • Cellular phenotype
  • Cellular phenotype
  • Fluorescent screening
  • In vivo confocal imaging
  • Trans-genic strain
  • Transparent skin mutant‐Casper (roy,nacre )
  • Zebrafish

ASJC Scopus subject areas

  • Medicine(all)

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