The hTH-GFP reporter rat model for the study of Parkinson's disease

PLoS One. 2014 Dec 2;9(12):e113151. doi: 10.1371/journal.pone.0113151. eCollection 2014.

Abstract

Parkinson disease (PD) is the second leading neurodegenerative disease in the US. As there is no known cause or cure for PD, researchers continue to investigate disease mechanisms and potential new therapies in cell culture and in animal models of PD. In PD, one of the most profoundly affected neuronal populations is the tyrosine hydroxylase (TH)-expressing dopaminergic (DA) neurons of the substantia nigra pars compacta (SNpc). These DA-producing neurons undergo degeneration while neighboring DA-producing cells of the ventral tegmental area (VTA) are largely spared. To aid in these studies, The Michael J. Fox Foundation (MJFF) partnered with Thomas Jefferson University and Taconic Inc. to generate new transgenic rat lines carrying the human TH gene promoter driving EGFP using a 11 kb construct used previously to create a hTH-GFP mouse reporter line. Of the five rat founder lines that were generated, three exhibited high level specific GFP fluorescence in DA brain structures (ie. SN, VTA, striatum, olfactory bulb, hypothalamus). As with the hTH-GFP mouse, none of the rat lines exhibit reporter expression in adrenergic structures like the adrenal gland. Line 12141, with its high levels of GFP in adult DA brain structures and minimal ectopic GFP expression in non-DA structures, was characterized in detail. We show here that this line allows for anatomical visualization and microdissection of the rat midbrain into SNpc and/or VTA, enabling detailed analysis of midbrain DA neurons and axonal projections after toxin treatment in vivo. Moreover, we further show that embryonic SNpc and/or VTA neurons, enriched by microdissection or FACS, can be used in culture or transplant studies of PD. Thus, the hTH-GFP reporter rat should be a valuable tool for Parkinson's disease research.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Axons / metabolism
  • Corpus Striatum / metabolism
  • Corpus Striatum / pathology
  • Disease Models, Animal
  • Dopamine / genetics
  • Dopamine / metabolism*
  • Dopaminergic Neurons / metabolism
  • Dopaminergic Neurons / pathology
  • Gene Expression Regulation
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics*
  • Humans
  • Mice
  • Olfactory Bulb / metabolism
  • Parkinson Disease / genetics*
  • Parkinson Disease / metabolism
  • Parkinson Disease / physiopathology
  • Pars Compacta / metabolism*
  • Promoter Regions, Genetic / genetics
  • Rats
  • Tyrosine 3-Monooxygenase / genetics

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Tyrosine 3-Monooxygenase
  • Dopamine

Grants and funding

This work was generously supported by a grant from the Michael J. Fox Foundation to LI. The funders: Sonal S. Das, Audrey Dufour, Terina N. Martinez, and Kuldip D. Dave analyzed the data. In addition, Kuldip D. Dave conceived and designed the experiments, and contributed to writing the manuscript.