Jones S J, Gray C, Sakamaki H, Arora M, Boyde A, Gourdie R, Green C
Department of Anatomy and Developmental Biology, University College London, UK.
Anat Embryol (Berl). 1993 Apr;187(4):343-52. doi: 10.1007/BF00185892.
Polyclonal antisera to synthetic peptides matching sequences on the cytoplasmic regions of connexin-43, a gap junction protein first identified in rat heart, have been used to immunolabel gap junctions in the calvarial bone, maintained intact as in vivo, of 1- to 2-week-old rats. The specimens were examined in reflection and fluorescence modes by scanning laser confocal microscopy, and the numbers of gap junctions and their sizes estimated. The mean number of connexin-43 immunolabelled junctions per osteoblast (n = 65) was 15.3 (SD +/- 4.5). The mean length of 227 junctions, selected for the sharpness of the image of the fluorescent spot, was 0.67 micron (SD +/- 0.18; range 0.37-1.29 microns) and their mean area 0.26 micron2 (SD +/- 0.145; range 0.075-0.93 micron2); these probably fell within the upper half of the total size range. Gap junctions were detected between preosteoblasts, osteoblasts, osteocytes and chondrocytes, and between these juxtaposed cell types. In addition, connexin-43 immunolabelled junctions were found between some osteoclasts and overlying mononuclear cells at active sites of resorption.
针对与连接蛋白43胞质区域序列匹配的合成肽制备的多克隆抗血清,已用于免疫标记1至2周龄大鼠颅骨中的缝隙连接,颅骨如在体内一样保持完整。通过扫描激光共聚焦显微镜以反射和荧光模式检查标本,并估计缝隙连接的数量及其大小。每个成骨细胞(n = 65)上连接蛋白43免疫标记连接的平均数量为15.3(标准差±4.5)。为荧光点图像的清晰度而选择的227个连接的平均长度为0.67微米(标准差±0.18;范围0.37 - 1.29微米),其平均面积为0.26平方微米(标准差±0.145;范围0.075 - 0.93平方微米);这些可能处于总大小范围的上半部分。在成骨前体细胞、成骨细胞、骨细胞和软骨细胞之间,以及这些并列的细胞类型之间检测到了缝隙连接。此外,在一些破骨细胞与吸收活跃部位的上层单核细胞之间发现了连接蛋白43免疫标记的连接。