The Green Fluorescent Protein is an Efficient Biological Marker for Cardiac Myocytes

M. Xian, N. Honbo, J. Zhang, C.-C. Liew, J. S. Karliner and Y.-F. C. Lau. The Green Fluorescent Protein is an Efficient Biological Marker for Cardiac Myocytes. Journal of Molecular and Cellular Cardiology (1999) 31, 2155–2165. There is a need for a non-toxic marker for cardiac myocytes in studies of cardiac development and in experimentally induced pathophysiologic states in adult animals. We investigated the possibility of using the enhanced green fluorescent protein (EGFP) gene as such a biological marker for cardiac myocytes in both whole animal and cell culture systems. Several lines of transgenic mice were constructed harboring an EGFP gene directed by a 2.38-kb promoter fragment of the hamsterβ -myosin heavy chain gene. The transgene was preferentially expressed in the cardiac progenitor cells of embryos at E7.5, a developmental stage that precedes the formation of the cardiomyotube. It was specifically expressed in the cardiomyotube and myotomes along the somites of embryos at E8.5. The EGFP transgene expression continued in the heart throughout gestation and became very intense at birth. When neonatal cardiac cells were fractionated into myocytes and non-myocytes by a differential plating procedure, only myocytes from the transgenic mice showed specific green fluorescence of the transgene product that can be used as a marker for flow cytometry analysis. Although the expression levels were heterogeneous, EGFP expression persisted in the hearts of postnatal animals. In addition to the heart, some skeletal and smooth muscles from transgenic animals also expressed the transgene. The transgenic mice were healthy and had a normal life span, identical to their non-transgenic littermates. These results demonstrate that EGFP is an efficient non-toxic biological marker for cardiac myocytes.