title: Seeing the trees in the forest: selective electroporation of adipocytes within adipose tissue.
abstract: Electroporation has been recently adapted for the transfer of macromolecules into cells of tissues in vivo. Although mature adipocytes constitute <20% of cells residing in adipose tissue, we hypothesized that fat cells might be susceptible to selective electrotransfer of plasmid DNA owing to their large size relative to other cells in the tissue. Results demonstrate the feasibility of electroporating DNA into mature fat cells with >99% selectivity over other cells in the tissue. Further experiments used the "adiporation" technique to image the subcellular targeting of fluorescent bioreporter molecules to the nucleus, mitochondria, and lipid droplets of adipocytes within intact adipose tissue. Finally, we utilized fluorescent bioreporters to examine the effects of constitutive activation of the beta-adrenergic signaling pathway in adipocytes. These results demonstrate that overexpression of rat beta1-adrenergic receptors alters the cellular morphology of white adipocytes in a fashion that mimics the effects of systemic infusion of beta3-adrenergic receptor agonists. Hallmarks of the altered morphology include pronounced fragmentation of the single lipid droplet, repositioning of the nucleus, and induction of mitochondrial biogenesis. These results indicate that activation of beta-adrenergic signaling within adipocytes is sufficient to induce a phenotype that resembles typical brown adipocytes and suggest that in vivo electroporation will allow molecular dissection of the mechanisms involved.
authors.affiliation: Center for Integrative Metabolic and Endocrine Research, Department of Pathology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. jgranne@med.wayne.edu
authors.forename: James G
authors.initials: JG
authors.lastname: Granneman
authors.forename: Pipeng
authors.initials: P
authors.lastname: Li
authors.forename: Yuyan
authors.initials: Y
authors.lastname: Lu
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Tilak
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D030161:Avian Proteins; D002352:Carrier Proteins / Q000502:physiology; D048429:Cell Size; D004247:DNA; D004268:DNA-Binding Proteins / Q000502:physiology; D018274:Electroporation; D005240:Feasibility Studies; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000648:ultrastructure; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; D000070778:Perilipin-1; D010750:Phosphoproteins / Q000235:genetics; D051381:Rats; D018342:Receptors, Adrenergic, beta-1 / Q000378:metabolism / Q000502:physiology; D015398:Signal Transduction; D017154:Stromal Cells / Q000166:cytology; D014162:Transfection
chemical_list: D030161:Avian Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D007202:Indicators and Reagents; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins; D000070778:Perilipin-1; D010750:Phosphoproteins; C085646:RBF-1 protein, Gallus gallus; D018342:Receptors, Adrenergic, beta-1; D049452:Green Fluorescent Proteins; D004247:DNA; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C434934:trans-2-enoyl-CoA reductase (NADPH)