Carlson E C, Kenney M C
J Morphol. 1982 Feb;171(2):195-211. doi: 10.1002/jmor.1051710207.
Freshly harvested kidneys from New Zealand white rabbits, Sprague-Dawley white rats, rhesus monkeys, and transplant-quality human kidneys were used in this study. Minced renal cortical tissue blocks (less than 2 mm3) were treated with 1 mM EDTA, 3% Triton X-100, 0.025% DNAse, and 4% sodium deoxycholate in an effort to remove all cellular elements and leave the extracellular matrix (ECM) intact. These preparations showed remarkable structural preservation and all components of the ECM, including basement membranes (BMs), maintained their in vivo histoarchitectural relationships. By light microscopy, at least four major BM types were recognizable, including Bowman's capsular BM (BCBM), tubular BM (TBM), glomerular BM (GBM), and peritubular capillary BM (PTCBM). Scanning electron microscopy demonstrated that, despite the lack of supporting interstitium, GBMs in human, monkey, and rat (and rabbit to a lesser degree) exhibit intrinsic structural rigidity such that their convoluted spheroidal shapes are maintained following cell removal. Transmission electron microscopy showed that major BM types are morphologically heterogeneous and vary markedly within and between species. Randomized measurements showed that isolated BM thicknesses (lamina densa only) compared favorably with those reported in cellular preparations. Mean thicknesses of GBMs were within normal ranges in all species with or without power transformations to reduce right-sided skew of distribution curves. In all species, thickness of BCBM greater than TBM greater than GBM greater than PTCBM. The striking morphologic heterogeneity of major BM types demonstrated in the acellular renal cortex is not surprising in view of recent biochemical analyses that show that BMs derived from different sources are compositionally disparate. We conclude that BMs should be evaluated and characterized individually and that morphologic definition of isolated BMs necessary prior to further analysis.
本研究使用了刚从新西兰白兔、Sprague-Dawley白鼠、恒河猴以及移植级别的人类肾脏中获取的新鲜肾脏。将肾皮质组织块切碎（小于2立方毫米），用1毫摩尔/升的乙二胺四乙酸（EDTA）、3%的 Triton X-100、0.025%的脱氧核糖核酸酶（DNAse）和4%的脱氧胆酸钠进行处理，以去除所有细胞成分并使细胞外基质（ECM）保持完整。这些制剂显示出显著的结构保存，并且ECM的所有成分，包括基底膜（BMs），都维持了它们在体内的组织架构关系。通过光学显微镜观察，至少可以识别出四种主要的BM类型，包括鲍曼囊基底膜（BCBM）、肾小管基底膜（TBM）、肾小球基底膜（GBM）和肾小管周围毛细血管基底膜（PTCBM）。扫描电子显微镜显示，尽管缺乏支持性间质，但人类、猴子和大鼠（兔子的程度较轻）的GBM表现出内在的结构刚性，以至于在细胞去除后仍能保持其盘绕的球形形状。透射电子显微镜显示，主要的BM类型在形态上是异质的，并且在物种内部和物种之间有显著差异。随机测量表明，分离出的BM厚度（仅指致密层）与细胞制剂中报道的厚度相当。在所有物种中，无论是否进行幂变换以减少分布曲线右侧的偏态，GBM的平均厚度都在正常范围内。在所有物种中，BCBM的厚度大于TBM大于GBM大于PTCBM。鉴于最近的生化分析表明，来自不同来源的BM在组成上存在差异，因此在无细胞肾皮质中显示出的主要BM类型惊人的形态异质性并不令人惊讶。我们得出结论，应该对BM进行单独评估和表征，并且在进一步分析之前，有必要对分离出的BM进行形态学定义。
