NDLI: A quantitative assay for lysosomal acidification rates in human osteoclasts.

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A quantitative assay for lysosomal acidification rates in human osteoclasts.

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A quantitative assay for lysosomal acidification rates in human osteoclasts.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Jensen, Vicki Kaiser Nosjean, Olivier Dziegiel, Morten Hanefeld Boutin, Jean A. Sørensen, Mette Grøndahl Karsdal, Morten Asser Henriksen, Kim
Description	Country affiliation: Denmark Author Affiliation: Jensen VK ( Nordic Bioscience A/S, Herlev, Denmark.)
Abstract	The osteoclast initiates resorption by creating a resorption lacuna. The ruffled border surrounding the lacunae arises from exocytosis of lysosomes. To dissolve the inorganic phase of the bone, the vacuolar adenosine triphosphatase, located in the ruffled border, pumps protons into the resorption lacunae. The electroneutrality of the lacunae is maintained by chloride transport through the chloride-proton antiporter chloride channel 7. Inhibition of either proton or chloride transport prevents bone resorption. The aims of this study were to validate the human osteoclastic microsome- based influx assay with respect to lysosomal acidification and assess whether it is a reliable test of a compound's ability to inhibit acidification. Investigated were the expression levels of the lysosomal acidification machinery, the activation of the assay by adenosine triphosphate, H(+) and Cl(-) dependency, the effect of valinomycin, inhibitor sensitivity, and the ion profile of the human osteoclast microsomes. The expression level of chloride channel 7 was increased in the human osteoclastic microsomes compared with whole osteoclasts. Acid influx was induced by 1.25 mM adenosine triphosphate. Further 1.1 µM valinomycin increased the acid influx by 129%. Total abrogation of acid influx was observed using both H(+) and Cl(-) ionophores. Finally, investigation of the anion profile demonstrated that Cl(-) and Br(-) are the preferred anions for the transporter. In conclusion, the acid influx assay based on microsomes from human osteoclasts is a useful tool for detection of inhibitors of the osteoclastic acidification machinery, and thus may aid the identification of effective drugs for osteoporosis that target the acid secretion by osteoclasts.
File Format	HTM / HTML
ISSN	1540658X
Issue Number	2
Volume Number	9
e-ISSN	15578127
Journal	ASSAY and Drug Development Technologies
Language	English
Publisher	Mary Ann Liebert, Inc.
Publisher Date	2011-04-01
Publisher Place	United States
Access Restriction	Subscribed
Subject Keyword	Discipline Analytical Discipline Pharmacology Discipline Chemistry Techniques Adenosine Triphosphatases Biological Assay Methods Chloride Channels Lysosomes Chemistry Osteoclasts Acids Antagonists & Inhibitors Metabolism Animals Standards Cattle Chromaffin Cells Enzyme Inhibitors Pharmacology Humans Leukocytes, Mononuclear Comparative Study Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Drug Discovery Molecular Medicine

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