In vivo imaging of MLC2v-luciferase, a cardiac-specific reporter gene expression in mice

Peter J. Gruber, Zijun Li, Hui Li, Diane Worrad, Bin Huang, Ibrahim Abdullah, Wenge Wang, Wafik El-Deiry, Victor A. Ferrari, Rong Zhou

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

A reporter or marker gene that is detectable by in vivo imaging permits longitudinal monitoring of certain fundamental biological processes (eg, differentiation) within the context of physiologically authentic environments. Tissue-specific expression of a reporter gene can be achieved when it is under the transcriptional control of a tissue-specific promoter. The objective of this study was to construct a plasmid vector containing firefly luciferase (Fluc) marker gene downstream of the promoter sequence of rat ventricular myosin light chain 2 (MLC2v); to detect the in vivo expression of this cardiac-specific reporter (MLC2v-Fluc) in the mouse heart by bioluminescent imaging; and to correlate the bioluminescent signal with postmortem luminometer assay. MLC2v-Fluc plasmid was generated by molecular cloning of 3 kb promoter sequence into a pGL3-Basic vector containing the Fluc reporter. Twenty μg of MLC2v-Fluc plasmid DNA in phosphate-buffered saline was directly injected into mouse myocardium through a midline sternotomy. At 1 week after injection, MLC2v-Fluc expression was detected by in vivo bioluminescent imaging in 60% of injected animals; the average in vivo signal intensity was (1.5 × 0.6) × 10 4 radiance (p/sec/cm 2/sr); in vivo signal was well above the detection threshold over 3 weeks after injection. In vivo bioluminescent signal is correlated (r 2 = 0.8) with the luminometer assay results from homogenized heart samples. The capability of noninvasive imaging of the MLC2v-Fluc in the heart will encourage applications that aim at monitoring and tracking the marker gene expression over time in cells undergoing cardiac differentiation.

Original languageEnglish (US)
Pages (from-to)1022-1028
Number of pages7
JournalAcademic Radiology
Volume11
Issue number9
DOIs
Publication statusPublished - Sep 1 2004

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Keywords

  • Cardiac ventricular myosin light chain 2 (MLC2v)
  • bioluminescence
  • cardiac
  • luciferase
  • reporter gene

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging

Cite this

Gruber, P. J., Li, Z., Li, H., Worrad, D., Huang, B., Abdullah, I., ... Zhou, R. (2004). In vivo imaging of MLC2v-luciferase, a cardiac-specific reporter gene expression in mice. Academic Radiology, 11(9), 1022-1028. https://doi.org/10.1016/j.acra.2004.05.022