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Putative fossil life in a hydrothermal system of the Dellen impact structure, Sweden
Stockholm University, Faculty of Science, Department of Geological Sciences. (Geokemi)
Naturhistoriska riksmuséet.
Stockholm University, Faculty of Science, Department of Geological Sciences. (Geokemi)
Stockholm University, Faculty of Science, Department of Geological Sciences. (Geologi)
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2010 (English)In: International Journal of Astrobiology, ISSN 1473-5504, E-ISSN 1475-3006, Vol. 9, no 3, 137-146 p.Article in journal (Refereed) Published
Abstract [en]

The role of muscarinic acetylcholine receptors (mAChRs) in regulating glucose uptake in L6 skeletal muscle cells was investigated. [H-3]-2-Deoxyglucose uptake was increased in differentiated L6 cells by insulin, acetylcholine, oxotremorine-M and carbachol. mAChR-mediated glucose uptake was inhibited by the AMPK inhibitor Compound C. Whole cell radioligand binding using [H-3]-N-methyl scopolamine chloride identified mAChRs in differentiated but not undifferentiated L6 cells and M-3 mAChR mRNA was detected only in differentiated cells. M-3 mAChRs are Gq-coupled, and cholinergic stimulation by the rnAChR agonists acetylcholine, oxotremorine-M and carbachol increased Ca2+ in differentiated but not undifferentiated L6 cells. This was due to muscarinic but not nicotinic activation as responses were antagonised by the muscarinic antagonist atropine but not the nicotinic antagonist tubocurarine. Western blotting showed that both carbachol and the AMPK activator AICAR increased phosphorylation of the AMPK alpha subunit at Thr172, with responses to carbachol blocked by Compound C and the CaMKK inhibitor STO609 but not by the PI3K inhibitor wortmannin. AICAR-stimulated AMPK phosphorylation was not sensitive to STO-609, confirming that this compound inhibits CaMKK but not the classical AMPK kinase LKB1. The TAK1 inhibitor (5Z)-7-oxozeaenol and the G(i) inhibitor pertussis toxin both failed to block AMPK phosphorylation in response to carbachol. Using CHO-K1 cells stably expressing each of the mAChR subtypes (M-1-M-4), it was determined that only the M-1 and M-3 mAChRs phosphorylate AMPK, confirming a G(q)-dependent mechanism. This study demonstrates that activation of M-3 mAChRs in L6 skeletal muscle cells stimulates glucose uptake via a CaMKK-AMPK-dependent mechanism, independent of the insulin-stimulated pathway.

Place, publisher, year, edition, pages
Cambridge: Cambridge University Press , 2010. Vol. 9, no 3, 137-146 p.
Keyword [en]
early Earth, early Mars, hydrothermal system, impact structure, microfossils
National Category
Other Earth and Related Environmental Sciences
Research subject
Geochemistry
Identifiers
URN: urn:nbn:se:su:diva-44360DOI: 10.1017/S147355041000011XISI: 000278258600013OAI: oai:DiVA.org:su-44360DiVA: diva2:361122
Projects
hydrotermal organisk geokemi
Note
authorCount :6Available from: 2010-11-08 Created: 2010-11-08 Last updated: 2017-12-12Bibliographically approved

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Neubeck, AnnaBroman, CurtHolm, Nils
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