Phelps P E, Barber R P, Houser C R, Crawford G D, Salvaterra P M, Vaughn J E
J Comp Neurol. 1984 Nov 1;229(3):347-61. doi: 10.1002/cne.902290306.
A monoclonal antibody to choline acetyltransferase (ChAT) has been used in an immunocytochemical study of the postnatal development of ChAT-containing neurons in cervical and thoracic spinal cord. Specimens from rat pups ranging in age from 1 to 28 days postnatal (dpn) were studied and compared with adult specimens (Barber et al., '84). The development of established cholinergic neurons, the somatic motoneurons and sympathetic preganglionic cells, has been described as has that of previously unidentified ChAT-positive neurons in the dorsal, intermediate, and central gray matter. Cell bodies of somatic and visceral motoneurons contained moderate amounts of ChAT-positive reaction product at birth that gradually increased in intensity until 14-21 dpn. The most intensely stained ChAT-positive neurons in 1-5-dpn specimens were named partition cells because this cell group extended from the central gray to an area dorsal to the lateral motoneurons, and thereby divided the spinal cord into dorsal and ventral halves. Partition cells were medium to large in size with 5-7 primary dendrites, and axons that, in fortuitous sections, could be traced into the ventrolateral motoneuron pools, the ventral funiculi, or the ventral commissure. Small ChAT-positive cells clustered around the central canal and scattered in laminae III-VI of the dorsal horn were detectable at birth. These neurons were moderately immunoreactive at 11-14 dpn and intensely ChAT positive by 21 dpn. The band of ChAT-positive terminal-like structures demonstrated in lamina III of adult specimens (Barber et al., '84) was first visible in 11-14-dpn specimens. By 28 dpn, both laminae I and III contained punctate bands that approximated the density of those observed in adult spinal cord. This investigation has demonstrated ChAT within individual neurons of developing spinal cord, and has identified a group of neurons, the partition cells, that exhibit intense ChAT-positive immunoreactivity earlier than any other putative cholinergic cells in spinal cord, including motoneurons. Another important observation has been that each ChAT-positive neuronal type achieves adult levels of staining intensity at different times during development. A likely explanation for this differential staining is that various groups of neurons acquire their mature concentration of ChAT molecules at different developmental stages. In turn, this may correlate with the maturation of cholinergic synaptic activity manifest by individual cells or groups of neurons.
一种针对胆碱乙酰转移酶(ChAT)的单克隆抗体已被用于对颈段和胸段脊髓中含ChAT神经元出生后发育的免疫细胞化学研究。研究了出生后1至28天(dpn)的大鼠幼崽标本,并与成年标本进行了比较(Barber等人,1984年)。已描述了成熟胆碱能神经元、躯体运动神经元和交感神经节前细胞的发育情况,以及背侧、中间和中央灰质中先前未鉴定的ChAT阳性神经元的发育情况。躯体和内脏运动神经元的细胞体在出生时含有适量的ChAT阳性反应产物,其强度逐渐增加,直至14 - 21 dpn。在1 - 5 dpn标本中染色最强烈的ChAT阳性神经元被命名为分隔细胞,因为这个细胞群从中央灰质延伸到外侧运动神经元背侧的区域,从而将脊髓分为背侧和腹侧两半。分隔细胞大小为中等至大型,有5 - 7条初级树突,其轴突在偶然的切片中可追踪到腹外侧运动神经元池、腹侧索或腹侧连合。出生时可检测到围绕中央管聚集并散布在背角III - VI层的小ChAT阳性细胞。这些神经元在11 - 14 dpn时免疫反应性中等,到21 dpn时ChAT呈强阳性。成年标本中在III层显示的ChAT阳性终末样结构带在11 - 14 dpn标本中首次可见。到28 dpn时,I层和III层都含有点状带,其密度与成年脊髓中观察到的相近。这项研究在发育中的脊髓单个神经元内证实了ChAT,并鉴定出一组神经元,即分隔细胞,它们比脊髓中任何其他假定的胆碱能细胞,包括运动神经元,更早地表现出强烈的ChAT阳性免疫反应性。另一个重要观察结果是,每种ChAT阳性神经元类型在发育过程中的不同时间达到成年水平的染色强度。这种差异染色的一个可能解释是,不同组的神经元在不同的发育阶段获得其成熟浓度的ChAT分子。反过来,这可能与单个细胞或神经元群体表现出的胆碱能突触活动的成熟有关。