Cellular and Molecular NeuroscienceResearch PaperCharacterization of Kiss1 neurons using transgenic mouse models
Research Highlights
▶Generation and validation of Kiss1-Cre mouse lines. ▶Kiss1 neurons in the arcuate nucleus (only) are direct target of leptin. ▶Kiss1 neurons coexpress melanocortin 4 receptor (MC4R). ▶Most of Kiss1 neurons in the AVPV/PeN (75%) are GABAergic. ▶Kiss1 neurons in the arcuate nucleus (95%) are glutamatergic.
Section snippets
Subjects
Adult male and female Kiss1-Cre, LepR-IRES-Cre/LacZ, MC4R-GFP and C57BL/6 mice were housed in the University of Texas Southwestern Medical Center Animal Resource Center, in a light- (12 h on/12 h off) and temperature- (21–23 °C) controlled environment. They were fed standard chow diet (Harlan Teklad Global Diet, Harlan Laboratories Inc., Indianapolis, IN, USA), unless otherwise mentioned and had free access to water. All experiments were carried out in accordance with the guidelines established
Production and validation of Kiss1-Cre mouse model
To generate transgenic mice that express Cre-recombinase driven by Kiss1 regulatory elements (Kiss1-Cre mouse model), we constructed two different Kiss1-Cre transgene-containing BACs (Fig. 1). We were successful in generating 14 different potential Kiss1-Cre founder mice. After crossing with reporter mice, we identified three lines with Cre activity in nuclei previously shown to express Kiss1 mRNA (Gottsch et al., 2004) (Fig. 2A, C, E, G). All three lines were originated from founders generated
Discussion
In the present study, we describe the generation and validation of a Kiss1-Cre transgenic mouse line (J2–4) which displayed Cre activity in areas known to express Kiss1 mRNA. We then generated Kiss1-reporter mice which allowed us to identify kisspeptin neurons in the mouse brain. We validated our mouse model through colocalization of the reporter gene and Kiss1 mRNA in the preoptic area (AVPV and Pe) and Arc, and confirmed the expression of Kiss1 in other sites, such as the medial nucleus of
Conclusion
In the present study we describe the generation of a novel Kiss1-Cre mouse model. This mouse model enabled us to generate reporters for Kiss1 neurons and to determine the chemical profiles of different populations of Kiss1 neurons. Future studies will take advantage of the Kiss1-specific expression of Cre recombinase to manipulate gene expression in Kiss1 neurons. This will allow us to further elucidate the mechanisms by which these neurons are influenced by changing metabolic states and by
Acknowledgments
We acknowledge the assistance of Robert Hammer and the UTSW Medical Center Transgenic Core Facility in generating the Kiss1-Cre transgenic animals, as well as of Angela Mobley and the UTSW Flow Cytometry Core Facility for the FACS procedure. We also thank Dr. Joyce Repa (UTSW) for Kiss1, Cyclophilin and GAD67 primers. We thank Dr. Jeffrey Friedman (Rockefeller University, New York) for kindly providing the LepR-IRES-Cre mice and Dr. Joel K. Elmquist (UTSW Medical Center, Dallas-TX) for kindly
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JMZ and CFE contributed equally to this work.