Yan Meijuan, Cheng Chun, Ding Fei, Jiang Jing, Gao Lingling, Xia Chunlin, Shen Aiguo
The Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, People's Republic of China.
Glycoconj J. 2008 Oct;25(7):685-701. doi: 10.1007/s10719-008-9129-5. Epub 2008 May 30.
Glycosylation is one of the most important post-translational modifications. It is clear that the single step of beta1,4-galactosylation is performed by a family of beta1,4-galactosyltransferases (beta1,4-GalTs), and that each member of this family may play a distinct role in different tissues and cells. beta1,4-GalT I and V are involved in the biosynthesis of N-linked oligosaccharides and play roles in sciatic nerve regeneration after sciatic nerve injury. In the present study, the expression of beta1,4-galactosyltransferase (beta1,4-GalT) I, V mRNAs and Galbeta1-4GlcNAc group were examined in rat gastrocnemius muscles after sciatic nerve crush and transection. Real time PCR revealed that beta1,4-GalT I and V mRNAs expressed at a high level in normal gastrocnemius muscles and decreased gradually from 6 h, reached the lowest level at 2 weeks, then restored gradually to relatively normal level at 4 weeks after sciatic nerve crush. In contrast, in sciatic nerve transection model, beta1,4-GalT I and V mRNAs decreased gradually from 6 h, and remained on a low level at 4 weeks in gastrocnemius muscles after sciatic nerve transection. In situ hybridization indicated that beta1,4-GalT I and V mRNAs localized in numerous myocytes and muscle satellite cells under normal conditions and at 4 weeks after sciatic nerve crush, and in a few muscle satellite cells at 4 weeks after sciatic nerve transection. Furthermore, lectin blotting showed that the expression level of the Galbeta1-4GlcNAc group decreased from 6 h, reached the lowest level at 2 weeks, and restored to relatively normal level at 4 weeks after sciatic nerve crush. RCA-I lectin histochemistry demonstrated that Galbeta1-4GlcNAc group localized in numerous membranes of myocytes and muscle satellite cells in normal and at 4 weeks after sciatic nerve crush, and in a few muscle satellite cells at 2 and 4 weeks after sciatic nerve transection. These results indicated that the expressions of beta1,4-GalT I, V mRNAs and Galbeta1-4GlcNAc group were involved in the process of denervation and reinnervation, which suggests that beta1,4-GalT I, V mRNAs and Galbeta1-4GlcNAc group may play an important role in the muscle regeneration.
糖基化是最重要的翻译后修饰之一。很明显,β1,4-半乳糖基化的单一步骤由β1,4-半乳糖基转移酶(β1,4-GalTs)家族执行,并且该家族的每个成员可能在不同组织和细胞中发挥不同作用。β1,4-GalT I和V参与N-连接寡糖的生物合成,并在坐骨神经损伤后的坐骨神经再生中发挥作用。在本研究中,检测了坐骨神经挤压和横断后大鼠腓肠肌中β1,4-半乳糖基转移酶(β1,4-GalT)I、V mRNA和Galβ1-4GlcNAc基团的表达。实时PCR显示,β1,4-GalT I和V mRNA在正常腓肠肌中高水平表达,在坐骨神经挤压后6小时开始逐渐下降,在2周时达到最低水平,然后在4周时逐渐恢复到相对正常水平。相比之下,在坐骨神经横断模型中,β1,4-GalT I和V mRNA在坐骨神经横断后6小时开始逐渐下降,并在4周时在腓肠肌中维持在低水平。原位杂交表明,β1,4-GalT I和V mRNA在正常条件下以及坐骨神经挤压后4周定位于大量肌细胞和肌肉卫星细胞中,而在坐骨神经横断后4周定位于少数肌肉卫星细胞中。此外,凝集素印迹显示,Galβ1-4GlcNAc基团的表达水平在坐骨神经挤压后6小时开始下降,在2周时达到最低水平,并在4周时恢复到相对正常水平。RCA-I凝集素组织化学表明,Galβ1-4GlcNAc基团在正常情况下以及坐骨神经挤压后4周定位于大量肌细胞膜和肌肉卫星细胞膜中,而在坐骨神经横断后2周和4周定位于少数肌肉卫星细胞膜中。这些结果表明,β1,4-GalT I、V mRNA和Galβ1-4GlcNAc基团的表达参与了去神经和再神经支配过程,这表明β1,4-GalT I、V mRNA和Galβ1-4GlcNAc基团可能在肌肉再生中发挥重要作用