Hansen-Smith F, Banker K, Morris L, Joswiak G
Department of Biological Science, Oakland University, Rochester, Michigan 48309-4401.
Microvasc Res. 1992 Jul;44(1):112-6. doi: 10.1016/0026-2862(92)90105-x.
Except in thin muscles, the analysis of muscle capillary depends on direct ultrastructural visualization or histochemical identification of the capillaries using stains for basement membrane or reactions for endothelial cell enzymes. The accuracy of existing histochemical methods for detecting capillaries has not been systematically tested, however. The purpose of the present study was to compare muscle capillarity determined by two methods currently in use, Griffonia simplicifolia I lectin (GSI) and ATPase, with two alternative capillary markers, Lycopersicon esculentum lectin (LEA) and MRC OX.43 antigen. Capillarity, expressed as capillaries around fibers (CAF), was determined in the soleus, extensor digitorum longus, and sternomastoid muscles from 4-month-old female rats. The GSI, LEA, and MRC OX.43 capillary markers gave identical results for each muscle, confirming their validity as cytochemical probes. In contrast, the capillary ATPase method gave significantly lower CAF determinations, with a differential effect of preincubations at pH 4.4 vs 4.0. These data suggest that capillary ATPase acid stability varies within the capillary bed. The use of one or more of the alternative capillary markers tested is suggested for studies of muscle capillarity to confirm that binding sites or enzyme activity used for a given probe is expressed under both control and experimental conditions.
除了在薄肌肉中，肌肉毛细血管的分析依赖于直接的超微结构可视化，或使用基底膜染色或内皮细胞酶反应对毛细血管进行组织化学鉴定。然而，现有的检测毛细血管的组织化学方法的准确性尚未得到系统测试。本研究的目的是比较目前使用的两种方法（简叶灰叶豆凝集素（GSI）和ATP酶）测定的肌肉毛细血管密度，与两种替代的毛细血管标记物（番茄凝集素（LEA）和MRC OX.43抗原）。以纤维周围的毛细血管（CAF）表示的毛细血管密度，在4月龄雌性大鼠的比目鱼肌、趾长伸肌和胸锁乳突肌中进行测定。GSI、LEA和MRC OX.43毛细血管标记物对每块肌肉给出了相同的结果证实了它们作为细胞化学探针的有效性。相比之下，毛细血管ATP酶法测定的CAF显著较低，在pH 4.4和4.0预孵育时有不同的影响。这些数据表明，毛细血管ATP酶的酸稳定性在毛细血管床内有所不同。建议在肌肉毛细血管研究中使用一种或多种经过测试的替代毛细血管标记物，以确认给定探针所使用的结合位点或酶活性在对照和实验条件下均有表达。
