TY - JOUR
T1 - Development of tg(Uas:Sec‐hsa.anxa5‐yfp,myl7:Rfp); casper(roy−/−,nacre−/−) transparent transgenic in vivo zebrafish model to study the cardiomyocyte function
AU - Rajpurohit, Surendra K.
AU - Gopal, Aaron
AU - Ye Mon, May
AU - Patel, Nikhil G.
AU - Arora, Vishal
N1 - Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/8
Y1 - 2021/8
N2 - 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.
AB - 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.
KW - Annexin‐5
KW - Cardiomyocyte
KW - Cellular phenotype
KW - Cellular phenotype
KW - Fluorescent screening
KW - In vivo confocal imaging
KW - Trans-genic strain
KW - Transparent skin mutant‐Casper (roy,nacre )
KW - Zebrafish
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U2 - 10.3390/cells10081963
DO - 10.3390/cells10081963
M3 - Article
C2 - 34440732
AN - SCOPUS:85115043011
SN - 2073-4409
VL - 10
JO - Cells
JF - Cells
IS - 8
M1 - 1963
ER -