Department of Kinesiology, University of Waterloo, Ontario, Canada.
Am J Physiol Cell Physiol. 2011 Oct;301(4):C841-9. doi: 10.1152/ajpcell.00409.2010. Epub 2011 Jun 22.
Sarcolipin (SLN) inhibits sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) pumps. To evaluate the physiological significance of SLN in skeletal muscle, we compared muscle contractility and SERCA activity between Sln-null and wild-type mice. SLN protein expression in wild-type mice was abundant in soleus and red gastrocnemius (RG), low in extensor digitorum longus (EDL), and absent from white gastrocnemius (WG). SERCA activity rates were increased in soleus and RG, but not in EDL or WG, from Sln-null muscles, compared with wild type. No differences were seen between wild-type and Sln-null EDL muscles in force-frequency curves or maximum rates of force development (+dF/dt). Maximum relaxation rates (-dF/dt) of EDL were higher in Sln-null than wild type across a range of submaximal stimulation frequencies, but not during a twitch or peak tetanic contraction. For soleus, no differences were seen between wild type and Sln-null in peak tetanic force or +dF/dt; however, force-frequency curves showed that peak force during a twitch and 10-Hz contraction was lower in Sln-null. Changes in the soleus force-frequency curve corresponded with faster rates of force relaxation at nearly all stimulation frequencies in Sln-null compared with wild type. Repeated tetanic stimulation of soleus caused increased (-dF/dt) in wild type, but not in Sln-null. No compensatory responses were detected in analysis of other Ca(2+) regulatory proteins using Western blotting and immunohistochemistry or myosin heavy chain expression using immunofluorescence. These results show that 1) SLN regulates Ca(2+)-ATPase activity thereby regulating contractile kinetics in at least some skeletal muscles, 2) the functional significance of SLN is graded to the endogenous SLN expression level, and 3) SLN inhibitory effects on SERCA function are relieved in response to repeated contractions thus enhancing relaxation rates.
肌浆网钙 ATP 酶(SERCA)的负调控因子肌联蛋白(SLN)。为了评估 SLN 在骨骼肌中的生理意义,我们比较了 Sln-/-和野生型小鼠的肌肉收缩和 SERCA 活性。野生型小鼠的 SLN 蛋白在比目鱼肌和红肌(RG)中表达丰富,在伸趾长肌(EDL)中表达较低,在白肌(WG)中不表达。与野生型相比,Sln-/-肌肉的比目鱼肌和 RG 的 SERCA 活性率增加,但 EDL 和 WG 肌肉没有增加。在力-频率曲线或最大力发展速率(+dF/dt)方面,野生型和 Sln-/-EDL 肌肉之间没有差异。在一系列亚最大刺激频率下,Sln-/-的 EDL 最大松弛率(-dF/dt)高于野生型,但在颤搐或最大强直收缩时则不然。对于比目鱼肌,在最大强直力或+dF/dt 方面,野生型和 Sln-/-之间没有差异;然而,力-频率曲线显示,在颤搐和 10-Hz 收缩时,Sln-/-的峰值力较低。与野生型相比,Sln-/-比目鱼肌的力-频率曲线变化导致几乎所有刺激频率下的力松弛速率更快。反复强直刺激比目鱼肌导致野生型的(-dF/dt)增加,但 Sln-/-则没有。通过 Western 印迹和免疫组化分析其他 Ca2+调节蛋白或免疫荧光分析肌球蛋白重链表达,未检测到代偿性反应。这些结果表明:1)SLN 调节 Ca2+-ATP 酶活性,从而调节至少一些骨骼肌的收缩动力学;2)SLN 的功能意义是分级的,与内源性 SLN 表达水平有关;3)SLN 对 SERCA 功能的抑制作用在反复收缩时得到缓解,从而提高松弛速率。