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Data from: The Chloride/proton Antiporter ClC3 is Critical for Normal Bone Formation in Mice

Blair, Harry (2024) Data from: The Chloride/proton Antiporter ClC3 is Critical for Normal Bone Formation in Mice. [Dataset] (Submitted)

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[img] Microsoft Excel (Dynamic bone formation data Acridine Xylenol Orange wild type vs CLC3 and CLC5 knockout mice)
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Abstract

Materials, methods, data, and figures for a paper currently under review.

Acid transport is required for bone synthesis by osteoblasts. The osteoblast basolateral surface extrudes acid by Na+/H+ exchange, but proton uptake is unclear. We found high expression of the Cl-/H+ exchanger ClC3 at the bone apical surface. In mammals ClC3 functions in intracellular vesicular chloride transport, but when we found Cl- dependency of H+ transport in osteoblast membranes, we queried whether ClC3 Cl-/H+ exchange functions in bone formation. We used ClC3 knockout animals, and closely-related ClC5 knockout animals: In vitro studies suggested that both ClC3 and ClC5 might support bone formation. Genotypes were confirmed by total exon sequences. Expression of ClC3, and to a lesser extent of ClC5, at osteoblast apical membranes was demonstrated by fluorescent antibody labeling and electron microscopy with nanometer gold labeling. Animals with ClC3 or ClC5 knockouts were viable. In ClC3 or ClC5 knockouts, bone formation decreased ~30% by calcein and xylenol orange labeling in vivo. In very sensitive micro-computed tomography, ClC5 knockout reduced bone relative to wild type, consistent with effects of ClC3 knockout, but was quantitatively smaller. Regrettably, ClC5-ClC3 double knockouts are not viable. We conclude that ClC3 has a direct role in bone formation with overlapping but probably slightly smaller effects of ClC5. The mechanism in mineral formation might include ClC H+ uptake, in contrast to ClC3 abd ClC5 function in cell vesicles or other organs.

Note for mass spectrometry data: These are the essential data for the MS evaluation of ClC3 expression. The KO expresses a shorter protein at about 60% of the level of the wild type. First two are wild type, second two are KO.

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Details

Item Type: Dataset
Status: Submitted
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Blair, Harryhcblair@pitt.eduhcblair0000-0002-1152-3718
Date: 11 January 2024
Type of Data: Text
Copyright Holders: University of Pittsburgh
Date Deposited: 07 Feb 2024 16:18
Last Modified: 01 Apr 2024 13:18
URI: http://d-scholarship.pitt.edu/id/eprint/45759

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