Bates C A, Stelzner D J
Developmental Biology Center, University of California at Irvine 92717.
Exp Neurol. 1993 Sep;123(1):106-17. doi: 10.1006/exnr.1993.1144.
It has been established that neonatal corticospinal (CS) axons are able to grow around lesions of the spinal cord early in neonacy (Bernstein and Stelzner, J. Comp. Neurol. 221:382-400; Firkins, Bates, and Stelzner, Exp. Neurol., 120:1-15). To determine if these corticospinal axons include regenerating as well as late developing axons a double-labeling paradigm is used in which CS neurons are retrogradely labeled from the cervical spinal cord by injections of fast blue (FB) on Postnatal Day (PND) 2, 4, or 10. Two days later, the FB is aspirated along with the left dorsal funiculus and the right hemicord (CHR). As adults, the animals receive an injection of diamidino yellow (DY) or rhodamine into the spinal cord caudal to the lesion site. Thus, FB neurons are those that originally projected to the spinal cord before the lesion and which survived axotomy, DY neurons are those whose axons reached the spinal cord after the lesion, and double-labeled neurons (FB/DY) are cells which projected to the spinal cord prior to the lesion and regenerated a spinal axon postlesion. Animals FB injected on PND 2 have a widespread distribution of FB-labeled neurons in cortex, including areas outside of sensorimotor cortex. These animals also had both DY- and FB/DY-labeled cells within sensorimotor cortex, indicating that the population of axons growing caudal to neonatal spinal lesions consists of both late growing and regenerating axons. In animals FB injected on PND 10, the FB neurons were all located in sensorimotor cortex. Very few DY and no FB/DY neurons were present. We have also looked at the topography of the CS neurons which project caudal to early spinal lesions. Rat pups received a CHR on PND 0, 3, 6, or 12. As adults, horseradish peroxidase was injected into the cervical or lumbar enlargement of the spinal cord and the distribution of labeled cells in the cerebral cortex was plotted and compared to normal and lesioned adult controls. In all experimental animals, the distribution of retrogradely labeled cells was restricted to the area containing CS projection neurons in the normal animal. This is despite the fact that the number of CS projection neurons is greatly reduced from normal and the normal pathway for CS axonal outgrowth has been completely disrupted by the neonatal lesion.
已经证实,新生期早期,新生大鼠的皮质脊髓(CS)轴突能够绕过脊髓损伤部位生长(伯恩斯坦和施特尔兹纳,《比较神经学杂志》221:382 - 400;菲尔金斯、贝茨和施特尔兹纳,《实验神经病学》,120:1 - 15)。为了确定这些皮质脊髓轴突是否包括再生轴突以及发育较晚的轴突,采用了一种双重标记模式,即通过在出生后第2天、第4天或第10天注射快蓝(FB),从颈脊髓对CS神经元进行逆行标记。两天后,将FB与左背侧索和右半脊髓(CHR)一并吸出。成年后,给动物在损伤部位尾侧的脊髓注射双脒基黄(DY)或罗丹明。因此,FB神经元是那些在损伤前最初投射到脊髓且在轴突切断后存活下来的神经元,DY神经元是那些其轴突在损伤后到达脊髓的神经元,而双重标记神经元(FB/DY)是那些在损伤前投射到脊髓且在损伤后再生出脊髓轴突的细胞。在出生后第2天注射FB的动物,FB标记的神经元在皮质中广泛分布,包括感觉运动皮质以外的区域。这些动物在感觉运动皮质内也有DY标记和FB/DY标记的细胞,这表明向新生脊髓损伤尾侧生长的轴突群体包括发育较晚的轴突和再生轴突。在出生后第10天注射FB的动物中,FB神经元都位于感觉运动皮质。几乎没有DY神经元,也没有FB/DY神经元。我们还研究了向早期脊髓损伤尾侧投射的CS神经元的拓扑结构。幼鼠在出生后第0天、第3天、第6天或第12天接受CHR。成年后,将辣根过氧化物酶注射到脊髓的颈膨大或腰膨大处,并绘制大脑皮质中标记细胞的分布图,与正常和损伤的成年对照进行比较。在所有实验动物中,逆行标记细胞的分布都局限于正常动物中含有CS投射神经元的区域。尽管CS投射神经元的数量比正常情况大幅减少,并且CS轴突生长的正常通路已被新生期损伤完全破坏。