QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online April 8, 2004
Journal of Experimental Biology 207, 1749-1756 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.00953

This Article Summary Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Ziegler, A. Articles by Bleher, R. Search for Related Content PubMed PubMed Citation Articles by Ziegler, A. Articles by Bleher, R. Social Bookmarking                         What's this?

Expression and polarity reversal of V-type H⁺-ATPase during the mineralization–demineralization cycle in Porcellio scaber sternal epithelial cells

Andreas Ziegler^1,*, Dirk Weihrauch², Monica Hagedorn¹, David W. Towle³ and Reiner Bleher¹

¹ Central Facility for Electron Microscopy, University of Ulm, Albert-Einstein-Allee 11, 89069 Ulm, Germany
² University of Osnabrück, Department of Animal Physiology, Barbarastraße 11, 49076 Osnabrück, Germany
³ Mount Desert Island Biological Laboratory, Salsbury Cove ME 04672, USA

View larger version (18K): [in a new window]   Fig. 1. Schematic representation of the epithelial H+, Ca2+ and transport during CaCO3 formation (A) and resorption (B).

View larger version (65K): [in a new window]   Fig. 3. Semiquantitative RT-PCR analysis of V-type H+-ATPase (VHA) mRNA abundance in sternal epithelial cells of Porcellio scaber during the mineralization–demineralization cycle. (A) Demonstration of template-dependent quantification of VHA B-subunit mRNA. (B) Relative expression of PMCA in the anterior (ASE) and posterior sternal epithelium (PSE).

View larger version (17K): [in a new window]   Fig. 2. Alignment of Porcellio scaber V-type H+-ATPase B-subunit amino acid fragment sequence (GenBank accession no. AY278992) with the corresponding isoforms of Callinectes sapidus (accession no. AF189780) and Manduca sexta (accession no. X64354). The black background indicates the agreement between the Porcellio and Callinectes or Manduca sequences.

View larger version (35K): [in a new window]   Fig. 4. Western blot analysis of homogenized sternal tissue of Porcellio scaber. The monoclonal antibody against the mouse monoclonal anti yeast V-type H+-ATPase B-subunit protein specifically binds to one band of about 54 kD (arrow).

View larger version (93K): [in a new window]   Fig. 5. Immunocytochemical localization of the V-type H+-ATPase (A–D and G–I) and Na+/K+-ATPase (E,F). (A,C,E,G) Phase contrast and (B,D,F,H,I) fluorescence micrographs of 0.7 µm thick cryosections of the anterior sternal epithelium. (A,B) Control stage. (C–F) Stage of CaCO3 deposit formation. (G–I) Stage of CaCO3 deposit resorption. (I) Staining without amplification step. bc, blood cell; bl, basal lamina; cl, cuticle; n, nucleus. Scale bars, 25 µm (A–H); 16 µm (I).

View larger version (189K): [in a new window]   Fig. 6. Electron micrographs of the anterior sternal epithelium of Porcellio scaber. (A) Overview of chemically fixed epithelial cells during formation of the CaCO3 deposits showing the distribution of the basolateral plasma membrane (bm) by extracellular invaginations. (B,C) Details of high-pressure frozen and freeze-substituted epithelial cells during the resorption of the CaCO3 deposits showing portasome coats (arrows) at the apical plasma membrane. bl, basal lamina; cl, cuticle; m, mitochondrion; n, nucleus. Scale bars, 2 µm (A); 200 nm (B–D).

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

Twitter