Division of Health Sciences, Graduate School of Medicine, Kanazawa University, Kanazawa 920-0942, Japan.
Neurochem Int. 2012 Aug;61(3):423-32. doi: 10.1016/j.neuint.2012.06.004. Epub 2012 Jun 16.
Unlike in mammals, fish retinal ganglion cells (RGCs) have a capacity to repair their axons even after optic nerve transection. In our previous study, we isolated a tissue type transglutaminase (TG) from axotomized goldfish retina. The levels of retinal TG (TG(R)) mRNA increased in RGCs 1-6weeks after nerve injury to promote optic nerve regeneration both in vitro and in vivo. In the present study, we screened other types of TG using specific FITC-labeled substrate peptides to elucidate the implications for optic nerve regeneration. This screening showed that the activity of only cellular coagulation factor XIII (cFXIII) was increased in goldfish optic nerves just after nerve injury. We therefore cloned a full-length cDNA clone of FXIII A subunit (FXIII-A) and studied temporal changes of FXIII-A expression in goldfish optic nerve and retina during regeneration. FXIII-A mRNA was initially detected at the crush site of the optic nerve 1h after injury; it was further observed in the optic nerve and achieved sustained long-term expression (1-40days after nerve injury). The cells producing FXIII-A were astrocytes/microglial cells in the optic nerve. By contrast, the expression of FXIII-A mRNA and protein was upregulated in RGCs for a shorter time (3-10days after nerve injury). Overexpression of FXIII-A in RGCs achieved by lipofection induced significant neurite outgrowth from unprimed retina, but not from primed retina with pretreatment of nerve injury. Addition of extracts of optic nerves with injury induced significant neurite outgrowth from primed retina, but not from unprimed retina without pretreatment of nerve injury. The transient increase of cFXIII in RGCs promotes neurite sprouting from injured RGCs, whereas the sustained increase of cFXIII in optic nerves facilitates neurite elongation from regrowing axons.
与哺乳动物不同,鱼类的视网膜神经节细胞(RGC)在视神经切断后仍有修复轴突的能力。在我们之前的研究中,我们从切断的金鱼视网膜中分离出一种组织型转谷氨酰胺酶(TG)。在神经损伤后 1-6 周,视网膜 TG(TG(R))mRNA 的水平升高,促进了体外和体内视神经的再生。在本研究中,我们使用特定的 FITC 标记底物肽筛选了其他类型的 TG,以阐明其对视神经再生的影响。该筛选显示,只有细胞凝血因子 XIII(cFXIII)的活性在神经损伤后立即增加。因此,我们克隆了 FXIII A 亚基(FXIII-A)的全长 cDNA 克隆,并研究了 FXIII-A 在金鱼视神经和视网膜再生过程中的表达随时间的变化。FXIII-A mRNA 在损伤后 1 小时即可在视神经的挤压部位检测到;在视神经中进一步观察到,并持续长期表达(损伤后 1-40 天)。产生 FXIII-A 的细胞是视神经中的星形胶质细胞/小胶质细胞。相比之下,FXIII-A mRNA 和蛋白的表达在 RGC 中上调时间较短(损伤后 3-10 天)。通过脂质体转染过表达 RGC 中的 FXIII-A 可诱导未经预处理的视网膜上的神经突显著生长,但对预处理过的视网膜则没有作用。损伤的视神经提取物的添加可诱导预处理过的视网膜上的神经突显著生长,但对未经预处理的视网膜则没有作用。RGC 中 cFXIII 的短暂增加促进了受损 RGC 中的神经突发芽,而视神经中 cFXIII 的持续增加则促进了再生轴突中的神经突伸长。