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Animal Models

Cdc2-like kinase 2 in the hypothalamus is necessary to maintain energy homeostasis

Abstract

Objective:

To investigate whether the Cdc2-like kinase 2 (CLK2) is expressed in hypothalamic neurons and if it is, whether the hypothalamic CLK2 has a role in the regulation of energy balance.

Subjects:

Swiss mice on chow or high-fat diet (HFD) and db/db mice on chow diet were used to address the role of CLK2 in the hypothalamus.

Results:

Hypothalamic CLK2Thr343 phosphorylation, which induces CLK2 activity, is regulated in vivo by refeeding, insulin and leptin, in a PI3K (phosphoinositide 3-kinase)-dependent manner. The reduction of CLK2 expression in the hypothalamus, by chronic pharmacological inhibition with TG003 or by chronic knockdown with small interfering RNA was sufficient to abolish the anorexigenic effect of insulin and leptin, to increase body weight, fat mass, food intake and to decrease energy expenditure in mice on chow. In contrast, CLK2Thr343 phosphorylation in the hypothalamus in response to insulin, leptin or refeeding was impaired in mice on HFD or in db/db mice. Chronic CLK2 inhibition in the hypothalamus was associated with a slight increase in the fasting blood glucose levels, reduction in PEPCK (phosphoenolpyruvate carboxykinase) expression in the liver and enhanced glucose production from pyruvate, suggesting a regulation of hepatic glucose production. Further, overexpressing CLK2 in the mediobasal hypothalami of mice on HFD or in db/db mice by adenovirus partially reversed the obese phenotype.

Conclusions:

Thus, our results suggest that protein CLK2 integrates some important hypothalamic pathways, and may be a promising molecule for new therapeutic approaches for obesity and diabetes.

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Acknowledgements

We are grateful for financial support from State University of Campinas (FUNCAMP), FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo): Auxílio Regular 2012/10338-6 and 2015/00343-0 and CEPID 2013/07607-8, São Paulo, Brazil and from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) Universal (481084/2013-4) and INCT (Instituto Nacional Ciência e Tecnologia de Obesidade e Diabetes; 573856/2008-7). The authors acknowledge the technical assistance of Luis Janeri, Jósimo Pinheiro, Dioze Guadagnini, Lígia Parreira Muniz and Ramon Henrick Zorzeto dos Santos from the Department of Internal Medicine, State University of Campinas. We thank Professor Armando Morais Ventura for kindly provide pShires vector. The present work received financial support from FAPESP (Fundação de Amparo a Pesquisa do Estado de São Paulo): Auxílio Regular 2012/10338-6 and CEPID 2013/07607-8, São Paulo, Brazil and from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) Universal (481084/2013-4) and INCT (Instituto Nacional Ciência e Tecnologia de Obesidade e Diabetes) (573856/2008-7).

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Correspondence to P O Prada.

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PGFQ designed, planned and performed all the experiments, analyzed data and wrote the manuscript; TMZ, ACS and LW performed experiments; JDJ and ICF performed and analyzed the immunohistochemistry assays; AHBM and ILC contributed with siRNA assays designing; FMS and PGFQ performed the adenovirus production and purification and reviewed the manuscript; POP directed the study, provided financial support and reviewed data analysis and the manuscript.

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Quaresma, P., Weissmann, L., Zanotto, T. et al. Cdc2-like kinase 2 in the hypothalamus is necessary to maintain energy homeostasis. Int J Obes 41, 268–278 (2017). https://doi.org/10.1038/ijo.2016.174

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