Gain of function AMP‐activated protein kinase γ3 mutation (AMPKγ3R200Q) in pig muscle increases glycogen storage regardless of AMPK activation

摘要

Chronic activation of AMP‐activated protein kinase (AMPK) increases glycogen content in skeletal muscle. Previously, we demonstrated that a mutation in the ryanodine receptor (RyR1^R615C) blunts AMPK phosphorylation in longissimus muscle of pigs with a gain of function mutation in the AMPK γ3 subunit (AMPK γ3^R200Q); this may decrease the glycogen storage capacity of AMPK γ3^R200Q + RyR1^R615C muscle. Therefore, our aim in this study was to utilize our pig model to understand how AMPK γ3^R200Q and AMPK activation contribute to glycogen storage and metabolism in muscle. We selected and bred pigs in order to generate offspring with naturally occurring AMPK γ3^R200Q, RyR1^R615C, and AMPK γ3^R200Q + RyR1^R615C mutations, and also retained wild‐type littermates (control). We assessed glycogen content and parameters of glycogen metabolism in longissimus muscle. Regardless of RyR1^R615C, AMPK γ3^R200Q increased the glycogen content by approximately 70%. Activity of glycogen synthase (GS) without the allosteric activator glucose 6‐phosphate (G6P) was decreased in AMPK γ3^R200Q relative to all other genotypes, whereas both AMPK γ3^R200Q and AMPK γ3^R200Q + RyR1^R615C muscle exhibited increased GS activity with G6P. Increased activity of style="fixed-case">GS with G6P was not associated with increased abundance of style="fixed-case">GS or hexokinase 2. However, style="fixed-case">AMPK γ3^R200Q enhanced style="fixed-case">UDP‐glucose pyrophosphorylase 2 ( style="fixed-case">UGP2) expression approximately threefold. Although style="fixed-case">UGP2 is not generally considered a rate‐limiting enzyme for glycogen synthesis, our model suggests that style="fixed-case">UGP2 plays an important role in increasing flux to glycogen synthase. Moreover, we have shown that the capacity for glycogen storage is more closely related to the style="fixed-case">AMPK γ3^R200Q mutation than activity.