Patch-clamp capacitance measurements: new insights into the endocytic uptake of transferrin

Blood Cells Mol Dis. 2002 Nov-Dec;29(3):459-64. doi: 10.1006/bcmd.2002.0584.

Abstract

Since its introduction by Neher and Sakmann in 1976 the patch-clamp technique has been extensively used to study processes such as signalling and synaptic transmission, but also for monitoring endo- and exocytosis. Since biological membranes behave like electrical capacitors high-resolution measurements of membrane capacitance allow detection of small changes in membrane surface area that accompany exocytosis and endocytosis. We here describe our recent work on patch-clamp capacitance measurements in stably transfected HeLa cells expressing HFE, the hereditary hemochromatosis gene product, under the control of a tetracycline-sensitive promotor. By means of whole-cell and cell-attached techniques we were able to reveal transferrin-induced decreases in membrane capacitance reflecting increased endocytosis at the single cell level. Moreover, cell-attached recordings revealed significant alterations in the formation of single endocytic vesicles. Time-resolved measurements of cell membrane capacitance provide a new methodological approach to study the endocytic uptake of transferrin and its regulation by HFE, the hereditary hemochromatosis protein.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Cytoplasmic Vesicles / metabolism
  • Endocytosis / physiology*
  • HeLa Cells
  • Hemochromatosis Protein
  • Histocompatibility Antigens Class I / metabolism
  • Humans
  • Iron / metabolism
  • Membrane Proteins / metabolism
  • Patch-Clamp Techniques / methods
  • Transferrin / metabolism*

Substances

  • HFE protein, human
  • Hemochromatosis Protein
  • Histocompatibility Antigens Class I
  • Membrane Proteins
  • Transferrin
  • Iron