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1 July 2004 Leptin-Receptor Gene Transfer into the Arcuate Nucleus of Female Fatty Zucker Rats Using Recombinant Adeno-Associated Viral Vectors Stimulates the Hypothalamo-Pituitary-Gonadal Axis
Erin Keen-Rhinehart, Satya P. Kalra, Pushpa S. Kalra
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Abstract

Fatty fa/fa Zucker rats with a missense mutation in the leptin receptor (OB-R) are obese and infertile with prolonged estrous cycles. To determine whether their reproductive deficits could be corrected by OB-R installation, we employed viral vectors to introduce the OB-R gene into either the arcuate nucleus (ARC) or the paraventricular nucleus (PVN) of the hypothalamus, sites of OB-R expression in wild-type rats. Recombinant adeno-associated viral (rAAV) vectors encoding the human leptin-receptor gene (rAAV-OB-Rb) were microinjected intraparenchymally to produce doxycycline-regulatable OB-R gene expression. Expression of the OB-R gene in the ARC and PVN was verified using reverse transcription-polymerase chain reaction. Expression of OB-R in the ARC, but not in the PVN, resulted in normalization of estrous cycle length, increased ovarian follicular development, and decreased serum progesterone levels. Compared to saline-injected rats, hypothalamic expression of neuropeptide Y (NPY) and pro-opiomelanocortin were decreased in ARC rAAV-OB-Rb-injected rats. Parallel decreases were noted in NPY and β-endorphin (β-END) concentrations in the hypothalamus, whereas luteinizing hormone-releasing hormone (LHRH) levels increased. These studies showed that rAAV vectors can be successfully used to install functional OB-R in the hypothalamus for extended periods. The resultant stimulation of the hypothalamo-pituitary-gonadal (HPG) axis in ARC-injected rats was probably brought about by the observed decreases in NPY and β-END, which inhibit hypothalamic LHRH. Because these changes were seen in ARC-injected, but not in PVN-injected, rats, the results suggest that the ARC may be the primary site where leptin acts to regulate the HPG axis.

Erin Keen-Rhinehart, Satya P. Kalra, and Pushpa S. Kalra "Leptin-Receptor Gene Transfer into the Arcuate Nucleus of Female Fatty Zucker Rats Using Recombinant Adeno-Associated Viral Vectors Stimulates the Hypothalamo-Pituitary-Gonadal Axis," Biology of Reproduction 71(1), 266-272, (1 July 2004). https://doi.org/10.1095/biolreprod.103.025858
Received: 11 December 2003; Accepted: 1 February 2004; Published: 1 July 2004
KEYWORDS
gonadotropin-releasing hormone
hypothalamic hormones
leptin
leptin receptor
neuropeptide Y
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