Watson-Wright W M, Wilkinson M
Muscle Nerve. 1986 Jun;9(5):416-22. doi: 10.1002/mus.880090506.
A new procedure is presented that characterizes the specific binding of the beta-adrenergic antagonist, [3H]CGP-12177, to thick (1 mm) slices from fast-twitch [extensor digitorum longus (EDL)] and slow-twitch (soleus) mouse skeletal muscle. Binding is reversible, saturable, stereospecific, of high affinity, and subject to agonist-induced desensitization, indicating that it is to beta-adrenoreceptors and not to other sites. In both muscles, the majority of specific binding is to the beta 2-receptor subtype. Bmax is approximately twice as high in the soleus (5.64 +/- 0.52 fmol/mg wet weight) as in the EDL (2.66 +/- 0.29 fmol/mg wet weight) (P less than 0.05), whereas affinity is higher in the fast-twitch (Kd = 0.30 +/- 0.08 nM) than the slow-twitch muscle (Kd = 0.45 +/- 0.08 nM). The minimal tissue disruption associated with this procedure, as well as its speed, simplicity and relatively low cost, suggest that the slice preparation may prove to be invaluable for the future study of beta-adrenergic receptor binding and associated responses in skeletal muscle.
本文介绍了一种新方法,该方法用于表征β-肾上腺素能拮抗剂[3H]CGP - 12177与来自快速收缩[趾长伸肌(EDL)]和慢速收缩(比目鱼肌)小鼠骨骼肌的厚(1毫米)切片的特异性结合。结合是可逆的、可饱和的、立体特异性的、高亲和力的,并且会受到激动剂诱导的脱敏作用,这表明它是与β-肾上腺素受体结合,而非其他位点。在这两种肌肉中,大部分特异性结合是与β2-受体亚型。比目鱼肌中的Bmax(最大结合容量)(5.64±0.52 fmol/mg湿重)约为EDL(2.66±0.29 fmol/mg湿重)的两倍(P<0.05),而快速收缩肌肉中的亲和力(Kd = 0.30±0.08 nM)高于慢速收缩肌肉(Kd = 0.45±0.08 nM)。该方法所涉及的组织损伤最小,且具有速度快、操作简单和成本相对较低的特点,这表明切片制备可能对未来骨骼肌中β-肾上腺素能受体结合及相关反应的研究具有极高的价值。