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Functional and structural characterization of endosomes from toad bladder epithelial cells.

M L Zeidel¹, T Hammond, B Botelho

¹Department of Veterans Affairs Medical Center, West Roxbury, Massachusetts 02132.

The American Journal of Physiology

|July 11, 1992

Summary

This study distinguishes toad bladder endosome subpopulations, identifying water channel-containing vesicles (WCV) responsible for antidiuretic hormone (ADH)-sensitive water transport and proton permeability.

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Area of Science:

Cell Biology
Membrane Transport
Physiology

Background:

Endosome heterogeneity complicates functional studies in toad bladder.
Distinguishing water channel-containing vesicles (WCV) from other endosomes is crucial.

Purpose of the Study:

To identify and characterize WCV and other endosome subpopulations.
To determine the role of ADH in regulating endosome water permeability.

Main Methods:

Flow cytometry to label and identify endosomes.
Centrifugation to separate endosome subpopulations (IP and HSP).
Permeability assays (water, proton) and structural analysis.

Main Results:

ADH-induced IP endosomes are purified WCV with high water permeability, sensitive to mercurials.

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HSP endosomes exhibit low permeability and contain constitutive, ADH-insensitive proteins.

ADH IP endosomes show high proton permeability, suggesting ADH water channels conduct protons.

Conclusions:

Toad bladder contains distinct ADH-sensitive (WCV) and constitutive endosome populations.
The ADH water channel facilitates both water and proton transport.
Multivesicular bodies do not contain functional water channels.