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Related Experiment Videos

Insulin uptake and processing by cultured mouse glomerular endothelial cells

R Rabkin1, T Tsao, S J Elliot

  • 1Stanford University School of Medicine, Department of Veterans Affairs Medical Center, Palo Alto.

The American Journal of Physiology
|August 1, 1993
PubMed
Summary

Glomerular endothelial cells (GEC) specifically bind and degrade insulin. These cells internalize insulin, with some degradation occurring within the cell, a process influenced by insulin and chloroquine.

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Area of Science:

  • Endocrinology
  • Cell Biology
  • Nephrology

Background:

  • Endothelial cells (EC) transport insulin but have low insulin degradation activity.
  • Vascular endothelial cells show heterogeneity, and the kidney is a primary site for insulin breakdown.

Purpose of the Study:

  • To investigate insulin processing by glomerular endothelial cells (GEC).
  • To understand the specific binding, degradation, and internalization mechanisms of insulin by GEC.

Main Methods:

  • Incubation of GEC with 125I-labeled insulin.
  • Measurement of insulin degradation using trichloroacetic acid (TCA) solubility.
  • Separation of membrane-bound and internalized insulin via pH changes.
  • Analysis of internalized insulin fate and release.

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  • High-performance liquid chromatography (HPLC) to identify insulin metabolites.

    • Main Results:

    • GEC specifically bind insulin, with degradation inhibited by excess insulin or ACTH.
    • Approximately 42% of 125I-insulin was degraded by GEC within 90 minutes.
    • In steady state, 43% of cell-associated insulin was membrane-bound and 57% internalized.
    • Internalized insulin was released as both TCA-soluble and TCA-insoluble products.
    • Chloroquine increased insulin release but decreased degradation, inhibiting intermediate formation.

    Conclusions:

    • GEC actively process insulin, involving specific binding, internalization, and degradation.
    • Insulin degradation by GEC is a regulated process, influenced by hormonal factors and potentially lysosomal pathways.
    • GEC contribute to renal insulin metabolism, with implications for understanding insulin clearance and action in the kidney.