Barrey E, Valette J P, Jouglin M, Picard B, Geay Y, Robelin J
INRA, station de génétique quantitative et appliquée, Jouy-en-Josas, France.
Reprod Nutr Dev. 1995;35(6):619-28. doi: 10.1051/rnd:19950602.
The content in slow and fast myosin heavy chains (MHC 1 and MHC 2) of 5 equine muscles was determined using an enzyme-linked immunosorbent assay. The results obtained with this immunoenzymatic method were compared with complementary techniques: electrophoresis and immunohistochemistry. Slices of masseter, diaphragm, tensor faciae latae, semitendinosus and cutaneus trunci were obtained from a 12-year-old saddle horse after slaughter. Muscular proteins were specifically extracted to be analysed by ELISA. The technique used 2 complimentary monoclonal antibodies (MAb). MAb 1 was prepared from a human atrium specimen that reacted specifically against MHC 1. Mab 2 was prepared from myosin of rabbit psoas muscle and reacted against MHC 2. The masseter muscle contained solely MHC 1 (100%) and this was confirmed by electrophoresis and immunohistochemistry. By contrast, the cutaneus trunci was very poor in MHC 1 (1.3%) and was entirely composed of MHC 2 (98.7%) which was confirmed by the other techniques. The diaphragm, tensor fasciae latae and semitendinosus contained 89, 40 and 2% of MHC 1, respectively. It was concluded that this ELISA method made it possible to measure a wide range of MHC contents in equine muscles with a good reproducibility. The results were consistent with those of the other fibre typing techniques. Moreover, this immunoenzymatic method is less time consuming than histological techniques and therefore offers new perspectives for muscle fibre typing in the horse.
采用酶联免疫吸附测定法测定了5种马肌肉中慢肌球蛋白重链和快肌球蛋白重链（MHC 1和MHC 2）的含量。将这种免疫酶法获得的结果与互补技术（电泳和免疫组织化学）进行了比较。从一匹12岁的鞍马屠宰后获取咬肌、膈肌、阔筋膜张肌、半腱肌和躯干皮肌的切片。特异性提取肌肉蛋白用于酶联免疫吸附测定分析。该技术使用了2种互补的单克隆抗体（MAb）。MAb 1由人心房标本制备，可特异性与MHC 1反应。Mab 2由兔腰大肌肌球蛋白制备，可与MHC 2反应。咬肌仅含有MHC 1（100%），这一点通过电泳和免疫组织化学得到了证实。相比之下，躯干皮肌中MHC 1含量极低（1.3%），完全由MHC 2组成（98.7%），其他技术也证实了这一点。膈肌、阔筋膜张肌和半腱肌中MHC 1的含量分别为89%、40%和2%。得出的结论是，这种酶联免疫吸附测定法能够以良好的重复性测量马肌肉中广泛的MHC含量。结果与其他纤维分型技术的结果一致。此外，这种免疫酶法比组织学技术耗时更少，因此为马的肌肉纤维分型提供了新的前景。
