Identification of hypothalamic genes implicated in the development of obesity in Psammomys obesus using differential display PCR.
The hypothalamus is a key central controller of energy homeostasis and is the source and/or site of action of many neuropeptides involved in this process. The aim of this study was to isolate hypothalamic genes differentially expressed between lean and obese Psammomys obesus, a polygenic animal model of obesity and type 2 diabetes. Differential display PCR was used to compare hypothalamic gene expression profiles of lean and healthy, obese and hyperinsulinemic, and obese, diabetic P. obesus in both the fed and fasted states. We conducted differential display with 180 separate primer combinations to amplify approximately 9,000 expressed transcripts. Sixty differentially expressed bands were excised. Taqman PCR was performed on 36 of these transcripts to confirm differential gene expression in a larger sample population. Of these 36 transcripts, 9 showed homology to known genes, and 27 were considered to be novel sequences. Gene expression profiles for two of these genes are presented here. In conclusion, differential display PCR was successfully used to isolate several transcripts that may be involved in the central regulation of energy balance. We are currently conducting numerous studies to further investigate the role of these genes in the development of obesity in P. obesus.
Trevaskis
James
J
Metabolic Research Unit, School of Health Sciences, Deakin University, Geelong, VIC 3217, Australia. jimt@deakin.edu.au
McMillan
Janine S
JS
Windmill
Kelly
K
Walder
Ken
K
Collier
Greg R
GR
D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D005223:Fats / Q000378:metabolism; D015870:Gene Expression; D005849:Gerbillinae / Q000235:genetics* / Q000378:metabolism; D007031:Hypothalamus / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D009765:Obesity / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods
D018076:DNA, Complementary; D005223:Fats