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Hypothalamic insulin expression remains unaltered after short-term fasting in female rats

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Abstract

Purpose

Our previous study showed that 6-h fasting increased insulin expression in the hypothalamus of male rats. We, therefore, wanted to examine if this phenomenon occurs in female rats and whether it depended on the estrus cycle phase.

Methods

Female rats in proestrus or diestrus were either exposed to 6-h fasting or had ad libitum access to food. The serum, cerebrospinal fluid, and hypothalamic insulin levels were determined using radioimmunoassay. The hypothalamic insulin mRNA expression was measured by RT-qPCR, while the hypothalamic insulin distribution was assessed immunohistochemically.

Results

Albeit the short-term fasting lowered circulating insulin, both hypothalamic insulin mRNA expression and hypothalamic insulin content remained unaltered. As for the hypothalamic insulin distribution, strong insulin immunopositivity was noted primarily in ependymal cells lining the upper part of the third ventricle and some neurons mainly located within the periventricular nucleus. The pattern of insulin distribution was similar between the controls and the females exposed to fasting regardless of the estrous cycle phase.

Conclusion

The findings of this study indicate that the control of insulin expression in the hypothalamus differs from that in the pancreatic beta cells during short-term fasting. Furthermore, they also imply that the regulation of insulin expression in the female hypothalamus is different from males but independent of the estrus cycle phase.

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Data availability

The datasets generated and/or analyzed during this study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors wish to thank Institute for Biological Research “Sinisa Stankovic”, Belgrade, Serbia, for the access to their microtome facility where coronal sections of the brain were made.

Funding

This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, under the contract number 451-03-68/2022-14/ 200178.

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Authors and Affiliations

  1. Department for Comparative Physiology and Ecophysiology, Institute for Physiology and Biochemistry Ivan Djaja, University of Belgrade-Faculty of Biology, Studentski trg 16, 11000, Belgrade, Serbia

    Tamara B. Dakic, Aleksandra A. Ruzicic, Tanja V. Jevdjovic, Iva V. Lakic, Jelena D. Djordjevic & Predrag Z. Vujovic

  2. Department of Cell and Tissue Biology, Institute for Zoology, University of Belgrade-Faculty of Biology, Studentski trg 16, 11000, Belgrade, Serbia

    Milica B. Markelic

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  1. Tamara B. Dakic
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Contributions

P.V. made the study conception and design. Material preparation, animal handling, data collection and analysis were performed by T.D. M.M. performed immunohistochemistry and interpreted the data. A.R. performed qPCR analysis. I.L. performed animal handling, collected CSF and interpreted the data. T.J. contributed to the interpretation of the results. The first draft of the manuscript was written by T.D. P.V. and M.M. performed review and editing. All authors provided critical feedback and helped shape the manuscript. J.Dj. reviewed the manuscript and obtained financial support. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Tamara B. Dakic.

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Conflict of interest

The authors declare no competing interests.

Ethics approval

All procedures were performed according to the Serbian Animals Welfare Law (No. 41/2009) which is in accordance with rules proposed by the Federation of European Laboratory Animal Science and Directive 2010/63/EU. The project was approved by the Ethics Committee of the Faculty of Biology, University of Belgrade (Permit No. EK-BF-2020/10) and Serbian Ministry of Agriculture, Forestry and Water Management, Veterinary Directorate (Permit No. 323-07-01936/2021-05).

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Dakic, T.B., Markelic, M.B., Ruzicic, A.A. et al. Hypothalamic insulin expression remains unaltered after short-term fasting in female rats. Endocrine 78, 476–483 (2022). https://doi.org/10.1007/s12020-022-03235-0

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