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Related Experiment Videos

Functional glucokinase isoforms are expressed in rat brain.

I Roncero1, E Alvarez, P Vázquez

  • 1Department of Biochemistry and Molecular Biology, Faculty of Medicine, Complutense University, Madrid, Spain.

Journal of Neurochemistry
|May 9, 2000
PubMed
Summary

This study identified glucokinase mRNA and protein in the rat brain, particularly the hypothalamus, aiding adaptation to blood glucose changes. Its co-expression with GLUT-2 suggests a role in neural glucose sensing.

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Area of Science:

  • Neuroendocrinology
  • Molecular Biology
  • Biochemistry

Background:

  • Glucokinase (GCK) mRNA has been identified in neuroendocrine cells, prompting further investigation into its role in the brain.
  • Understanding GCK expression and function in the brain is crucial for comprehending glucose regulation and adaptation.

Purpose of the Study:

  • To characterize glucokinase gene expression and protein activity in the rat brain, focusing on the hypothalamus.
  • To investigate the presence and function of glucokinase regulatory protein (GCKRP) mRNA in the rat brain.

Main Methods:

  • Reverse transcription-polymerase chain reaction (RT-PCR) to detect glucokinase mRNA isoforms and GCKRP mRNA.
  • Enzyme activity assays to determine kinetic properties (Km, product inhibition) of brain glucokinase.

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  • Western blot analysis to identify the molecular weight of the brain glucokinase protein.

    • Main Results:

    • Glucokinase mRNA, predominantly the B1 isoform, was detected in rat hypothalamus and pancreatic islets, with similar splicing patterns.
    • Brain glucokinase exhibited high apparent Km for glucose, lacked glucose-6-phosphate product inhibition, and contributed significantly to glucose phosphorylating activity in the hypothalamus and cerebral cortex.
    • RT-PCR confirmed the presence of glucokinase regulatory protein mRNA in the rat brain, similar to liver.

    Conclusions:

    • Glucokinase in the rat brain, especially the hypothalamus, plays a role in adapting to blood glucose fluctuations.
    • Co-expression of glucokinase and GLUT-2 in hypothalamic neurons suggests a critical function in neural glucose sensing and regulation.