Lee M T, Koebbe M J, O'Donovan M J
Department of Physiology and Biophysics, University of Iowa, Iowa City 52242.
J Neurosci. 1988 Jul;8(7):2530-43. doi: 10.1523/JNEUROSCI.08-07-02530.1988.
We have examined the development of synaptic connections between afferents and motoneurons in the lumbosacral spinal cord of the chick embryo between stages 28 and 39. The central projection of afferents was visualized following injection of dorsal root ganglia with HRP. Afferent fibers first entered the dorsal gray matter between stages 29 and 31. They grew in a ventrolateral direction, reaching motoneuron dendrites by stage 32. Quantitative analysis of axon numbers suggested that individual axons did not begin to branch extensively until they approached the lateral motor column at stage 36. Connectivity between afferents and motoneurons was assessed by stimulating dorsal roots or nerves supplying the femorotibialis muscle and recording the resulting motoneuron synaptic potentials intracellularly or from the cut ventral roots. At stages 37-39, low-intensity stimulation produced a short-latency positive potential that was followed at higher stimulus currents by slower positive potentials. All of these potentials were abolished in solutions that block chemical synaptic transmission (zero Ca2+/2 mM Mn2+). The early potential, which includes the monosynaptic EPSP produced by muscle afferents, persisted in the presence of the N-methyl-D-aspartate antagonist, 2-amino-5-phosphonovaleric acid (APV), but was largely eliminated by the more general excitatory amino acid antagonist, kynurenic acid. Therefore, in the chick, as in other species, a glutamate-like transmitter appears to be released at the synapses between muscle afferents and motoneurons. The APV-resistant potential was reduced in amplitude during bath application of the glycine and GABA antagonists, strychnine and picrotoxin, suggesting that it was composed of depolarizing inhibitory as well as excitatory components at these stages. The monosynaptic EPSP could be recorded in ventral roots as early as stages 32-33, when muscle afferents first grew into the vicinity of motoneuron dendrites. The EPSP in these young embryos was unaffected by picrotoxin and strychnine, but responded to APV and kynurenate in a manner similar to that at later stages. Between stages 28 and 32, only long-latency, slowly rising potentials could be evoked in the ventral roots by afferent activation. These potentials were abolished by superfusion with zero Ca2+/2 mM Mn2+, APV, or kynurenic acid, and could be revealed before stage 31 only by removing Mg2+ from the bath.(ABSTRACT TRUNCATED AT 400 WORDS)
我们研究了鸡胚在第28至39阶段腰荐脊髓中传入神经与运动神经元之间突触连接的发育情况。通过向背根神经节注射辣根过氧化物酶(HRP)来观察传入神经的中枢投射。传入纤维在第29至31阶段首次进入背侧灰质。它们向腹外侧方向生长,到第32阶段到达运动神经元树突。对轴突数量的定量分析表明,单个轴突直到第36阶段接近外侧运动柱时才开始广泛分支。通过刺激背根或供应股胫肌的神经并在细胞内或从切断的腹根记录由此产生的运动神经元突触电位,来评估传入神经与运动神经元之间的连接性。在第37至39阶段,低强度刺激产生一个短潜伏期的正电位,在较高刺激电流时随后会出现较慢的正电位。所有这些电位在阻断化学突触传递的溶液(零钙/2毫摩尔锰)中都会消失。早期电位,包括肌肉传入神经产生的单突触兴奋性突触后电位(EPSP),在存在N-甲基-D-天冬氨酸拮抗剂2-氨基-5-磷酸戊酸(APV)时持续存在,但在更通用的兴奋性氨基酸拮抗剂犬尿烯酸存在时基本消失。因此,在鸡中,与其他物种一样,在肌肉传入神经与运动神经元之间的突触处似乎释放一种类似谷氨酸的递质。在浴应用甘氨酸和GABA拮抗剂士的宁和印防己毒素期间,APV抗性电位的幅度降低,这表明在这些阶段它由去极化抑制性成分以及兴奋性成分组成。单突触EPSP最早可在第32至33阶段在腹根中记录到,此时肌肉传入神经首次长入运动神经元树突附近。这些幼胚中的EPSP不受印防己毒素和士的宁影响,但对APV和犬尿酸盐的反应方式与后期相似。在第28至32阶段之间,传入神经激活只能在腹根中诱发长潜伏期、缓慢上升的电位。这些电位通过用零钙/2毫摩尔锰、APV或犬尿烯酸进行灌流而消失,并且只有在第31阶段之前通过从浴中去除镁才能显示出来。(摘要截断于400字)