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Targeting of the Enhanced Green Fluorescent Protein Reporter to Adrenergic Cells in Mice

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Abstract

Adrenaline and noradrenaline are important neurotransmitter hormones that mediate physiological stress responses in adult mammals, and are essential for cardiovascular function during a critical period of embryonic/fetal development. In this study, we describe a novel mouse model system for identifying and characterizing adrenergic cells. Specifically, we generated a reporter mouse strain in which a nuclear-localized enhanced green fluorescent protein gene (nEGFP) was inserted into exon 1 of the gene encoding Phenylethanolamine n-methyltransferase (Pnmt), the enzyme responsible for production of adrenaline from noradrenaline. Our analysis demonstrates that this knock-in mutation effectively marks adrenergic cells in embryonic and adult mice. We see expression of nEGFP in Pnmt-expressing cells of the adrenal medulla in adult animals. We also note that nEGFP expression recapitulates the restricted expression of Pnmt in the embryonic heart. Finally, we show that nEGFP and Pnmt expressions are each induced in parallel during the in vitro differentiation of pluripotent mouse embryonic stem cells into beating cardiomyocytes. Thus, this new mouse genetic model should be useful for the identification and functional characterization of adrenergic cells in vitro and in vivo.

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Acknowledgments

This study was supported by funding from the NIH (HL78716) to S.N.E., and intramural NICHD funds to K.P.

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Authors and Affiliations

  1. Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 6900 Lake Nona Blvd, Orlando, FL, 32827-7047, USA

    Jixiang Xia, Namita Varudkar, Candice N. Baker, Ibrahim Abukenda, Celines Martinez & Steven N. Ebert

  2. Department of Pediatrics, Medstar-Georgetown University Hospital, Washington, DC, USA

    Aruna Natarajan

  3. Program in Genomics of Differentiation, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA

    Alexander Grinberg & Karl Pfeifer

Authors
  1. Jixiang Xia
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  2. Namita Varudkar
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  3. Candice N. Baker
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  4. Ibrahim Abukenda
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  5. Celines Martinez
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  6. Aruna Natarajan
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  7. Alexander Grinberg
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  8. Karl Pfeifer
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  9. Steven N. Ebert
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Corresponding author

Correspondence to Steven N. Ebert.

Electronic Supplementary Material

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12033_2012_9570_MOESM1_ESM.tif

Supplemental Figure 1. Control Pnmt +/+ mouse adrenal gland sections. (A) nEGFP expression (green). (B) Anti-GFP IF staining (red). (C) DAPI nuclear stain (blue). (D) Overlay of images obtained in panels A-C. Scale bar, 30 μm. Only the medulla region of the adrenal section is shown for comparison with positive staining observed in Fig. 5. (TIFF 2599 kb)

12033_2012_9570_MOESM2_ESM.tif

Supplemental Figure 2. Fluorescence expression and staining in control undifferentiated mESCs. (A) nEGFP expression (green). (B) Anti-GFP IF staining (red). (C) DAPI nuclear stain (blue). (D) Overlay of images obtained in panels A-C. Scale bar, 10 μm. (TIFF 4653 kb)

12033_2012_9570_MOESM3_ESM.tif

Supplemental Figure 3. Fluorescence expression and staining in cardiac-differentiated (7 + 5d) mESCs. (A) nEGFP expression (green). (B) Anti-GFP IF staining (red). (C) DAPI nuclear stain (blue). (D) Overlay of images obtained in panels A-C. Scale bar, 10 μm. (TIFF 1607 kb)

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Xia, J., Varudkar, N., Baker, C.N. et al. Targeting of the Enhanced Green Fluorescent Protein Reporter to Adrenergic Cells in Mice. Mol Biotechnol 54, 350–360 (2013). https://doi.org/10.1007/s12033-012-9570-3

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  • DOI: https://doi.org/10.1007/s12033-012-9570-3

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