LeBlanc A C, Földvári M, Spencer D F, Breckenridge W C, Fenwick R G, Williams D L, Mezei C
Department of Neurology, Mayo Clinic, Rochester, Minnesota 55905.
J Cell Biol. 1989 Sep;109(3):1245-56. doi: 10.1083/jcb.109.3.1245.
The expression of the apolipoprotein A-I (apo A-I) gene was investigated in the myelinating sciatic nerve. Hybridization analysis with an apo A-I cDNA probe obtained from a cDNA library of mRNA isolated from rapidly myelinating chick sciatic nerve indicated that apo A-I coding transcripts increase during development in the chick sciatic nerve in parallel with the increase of myelin lamellae. Substantial apo A-I-like immunoreactivity in chick sciatic nerve homogenates was detected by Western blotting. The amount of antigen increased from the 15-d embryonic stage to 1 d posthatch and then decreased. Two subcellular fractions corresponding to the cytoplasmic compartments were particularly enriched in apo A-I. apo A-I immunoreactivity was also found in highly purified myelin preparations. Immunohistochemical staining provided further evidence for the presence of apo A-I in the endoneurial compartment of the sciatic nerve. Electron microscopic examination of these fractions after negative staining showed the presence of spherical and disc-shaped particles resembling high density lipoproteins. The presence of apo A-I, cholesterol esters, phospholipids, and triacylglycerols in ultracentrifugal fractions corresponding to serum lipoproteins and the behavior of apo A-I on nondenaturing gradient gels implied that apo A-I was associated with lipid. Studies with short-term organ cultures of sciatic nerves from 1-d chicks strengthened the evidence for local synthesis and secretion of apo A-I and apo A-I-containing lipoproteins by this tissue. These results establish that the apo A-I gene is actively expressed in developing sciatic nerve during the period of rapid myelination. These findings support the hypothesis that apo A-I synthesized within the nerve participates in the local transport of lipids used in myelin biosynthesis.
对髓鞘形成期坐骨神经中的载脂蛋白A-I(apo A-I)基因表达进行了研究。用从快速髓鞘形成期鸡坐骨神经分离的mRNA的cDNA文库获得的apo A-I cDNA探针进行杂交分析表明,在鸡坐骨神经发育过程中,apo A-I编码转录本随着髓鞘板层的增加而平行增加。通过蛋白质免疫印迹法检测到鸡坐骨神经匀浆中有大量apo A-I样免疫反应性。抗原量从胚胎15天期增加到出壳后1天,然后下降。对应于细胞质区室的两个亚细胞组分中apo A-I特别富集。在高度纯化的髓鞘制剂中也发现了apo A-I免疫反应性。免疫组织化学染色为坐骨神经内膜区室中存在apo A-I提供了进一步证据。对这些组分进行负染后的电子显微镜检查显示存在类似于高密度脂蛋白的球形和盘状颗粒。对应于血清脂蛋白的超速离心组分中存在apo A-I、胆固醇酯、磷脂和三酰甘油,以及apo A-I在非变性梯度凝胶上的行为表明apo A-I与脂质相关。对1日龄雏鸡坐骨神经进行短期器官培养的研究进一步证明了该组织可局部合成和分泌apo A-I以及含apo A-I的脂蛋白。这些结果表明,在快速髓鞘形成期,apo A-I基因在发育中的坐骨神经中活跃表达。这些发现支持了这样一种假说,即神经内合成的apo A-I参与了髓鞘生物合成中所用脂质的局部运输。