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Am J Physiol Endocrinol Metab (October 14, 2003). doi:10.1152/ajpendo.00147.2003
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Submitted on April 4, 2003
Accepted on October 6, 2003

Expression profiling of the {gamma}-subunit isoforms of AMP-activated protein kinase suggests a major role for {gamma}3 in white skeletal muscle

Margit Mahlapuu1, Carina Johansson1, Kerstin Lindgren1, Goran Hjalm2, Brian R. Barnes3, Anna Krook3, Juleen R. Zierath3, Leif Andersson2, and Stefan Marklund2*

1 Arexis AB, Gothenburg, Sweden
2 Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Uppsala Biomedical Center, Uppsala, Sweden
3 Department of Surgical Sciences, Karolinska Institutet, Stockholm, Sweden

* To whom correspondence should be addressed. E-mail: Stefan.Marklund{at}bmc.uu.se.

Expression patterns of the three isoforms of the regulatory {gamma}-subunit of adenosine monophosphate-activated protein kinase (AMPK) were determined in various tissues from adult humans, mice and rats, as well as in human primary muscle cells. Real-time PCR based quantification of mRNA showed similar expression patterns in the three species and a good correlation with protein expression in mice and rats. The {gamma}3-isoform appeared highly specific to skeletal muscle, whereas {gamma}1 and {gamma}2 showed broad tissue distributions. Moreover, the proportion of white, type IIb fibers in the mouse and rat muscle samples, as indicated by real-time PCR quantification of Atp1b2 mRNA, showed a strong positive correlation to the expression of {gamma}3. In samples of white skeletal muscle, {gamma}3 clearly appeared to be the most abundant {gamma}-isoform. Differentiation of human primary muscle cells from myoblasts into multinucleated myotubes was accompanied by upregulation of {gamma}3-mRNA expression, while levels of {gamma}1 and {gamma}2 remained largely unchanged. However, even in these cultured myotubes, {gamma}2 was the most highly expressed isoform, indicating a considerable difference compared to adult skeletal muscle. Immunoblot analysis of mouse gastrocnemius and quadriceps muscle extracts precipitated with a {gamma}3-specific antibody showed that {gamma}3 was exclusively associated with the {alpha}2- and {beta}2-subunit isoforms. The observation that the AMPK {gamma}3-isoform is primarily expressed in white skeletal muscle, in which it is the predominant {gamma}-isoform, strongly suggests that {gamma}3 has a key role in this tissue.




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