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Am J Physiol Endocrinol Metab (December 23, 2003). doi:10.1152/ajpendo.00404.2003
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Submitted on September 9, 2003
Accepted on December 11, 2003

An Important Role for the V-Type (H+)-ATPase and the Golgi Apparatus in the Recycling of PTH/PTHrP Receptor

Hesham A W Tawfeek1* and Abdul B. Abou-Samra1

1 Endocrine Unit, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA

* To whom correspondence should be addressed. E-mail: tawfeek{at}helix.mgh.harvard.edu.

Our previous studies demonstrated that a green fluorescent protein (GFP)-tagged parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor stably expressed in LLCPK-1 cells undergoes agonist-dependent internalization into clathrin-coated pits. The sub-cellular localization of the internalized PTH/PTHrP receptor is not known. In the current study, we explore the intracellular pathways of the internalized PTH/PTHrP receptor. Using immuno-fluorescence and confocal microscopy, we show that the internalized receptors localize at a juxta-nuclear compartment identified as the Golgi apparatus. The receptors do not co-localize with lysosomes. Furthermore, whereas the internalized receptors exhibit rapid recycling, treatment with proton pump inhibitors (bafilomycin-A1 and concanamycin A) or brefeldin A, Golgi disrupting agents, reduces PTH/PTHrP receptor recycling. Together, these data indicate an important role for the vacuolar-type hydrogen-ATPase and the Golgi apparatus in post-endocytic PTH/PTHrP receptor recovery.




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