Free-flow electrophoretic analysis of endosome subpopulations of rat hepatocytes

Electrophoresis
I StefanerR Fuchs

Abstract

The separation of functional early and late endosomes from other cellular compartments by free-flow electrophoresis (FFE) has been previously demonstrated in nonpolarized cells. Here, using 125I-labeled anti-secretory component antibodies ([125I]SC Ab) and FITC-labeled asialoorosomucoid (FITC-ASOR) as markers of the transcytotic and lysosomal pathway, respectively, we demonstrate the separation of three distinct endosome subpopulations from polarized rat hepatocytes. Internalization of both markers at 16 degrees C resulted in their accumulation in a common endosome compartment, indicating that both the transcytotic and the lysosomal pathways are arrested in the sorting early endosome at temperatures below 20 degrees C. After chase of the markers from early endosomes into the transcytotic or the degradative route at 37 degrees C, transcytotic endosomes carrying [125I]SC Ab migrated with an electrophoretic motility between early and late endosomes while late endosomes labeled with FITC-ASOR were deflected more towards the anode than early endosomes. These data indicate that in rat hepatocytes, the transcytotic and lysosomal pathways utilize a common (i.e. early endosomes) and two distinct endosome subpopulations (i.e. transcytoti...Continue Reading

References

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Citations

Jun 2, 2005·Analytical and Bioanalytical Chemistry·Karen J OlsonEdgar A Arriaga
Nov 11, 2008·Wiener medizinische Wochenschrift·Isabella Ellinger, Renate Fuchs
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·C PasqualiL A Huber
Feb 20, 2010·Journal of Biomedicine & Biotechnology·Isabella Ellinger, Renate Fuchs
Jul 14, 1998·Electrophoresis·L Krivánková, P Bocek
Jul 12, 2002·The Journal of Gene Medicine·Christine GonçalvesPatrick Midoux
Jan 30, 2008·Current Protocols in Cell Biology·Renate Fuchs, Isabella Ellinger
Aug 30, 2008·Molecular Pharmaceutics·Lisa M BarefordPeter W Swaan

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