FURTHER OBSERVATIONS ON HIPPOCAMPAL-NEURONS IN DISPERSED CELL-CULTURE
FURTHER OBSERVATIONS ON HIPPOCAMPAL-NEURONS IN DISPERSED CELL-CULTURE
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DOI：
10.1002/cne.901870302
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发表时间：
1979-01-01
影响因子：
2.5
通讯作者：
COWAN, WM
中科院分区：
文献类型：
--
作者：
BANKER, GA;COWAN, WM
使用一种允许细胞以相对低的铺板密度维持长达3周的方法，在体外研究了来自18-20日龄大鼠胎儿的分离海马神经元突起的生长（Banker和Cown，1977）。大多数细胞在最初附着于基质（多聚赖氨酸或胶原）时为球形或卵形，但在24至48小时内，它们开始发出突起。在培养的第1周结束时，显著比例（约45%）与正常锥体细胞相似，具有或多或少的三角形索马体，从索马体的顶端出现的单个主导树突样突起，以及从细胞的对极出现的数个基底树突。比较这些树突状突起的长度与海马细胞在动物的大脑中处死的第4天出生后的一个变体的Golgi-Cox方法浸渍表明，在某些情况下，在体外的过程形成的速度接近，在体内和一般形式的神经元是非常像的未成熟的锥体细胞。在培养1周后，可以识别第二种类型的过程。它们往往比那些被鉴定为树突的更细;它们的直径相对均匀，经常以直角发出分支，并且在EM中可以看到在较大的突起和细胞体上形成突触。这些轴突样突起与正常锥体细胞的轴突在两个重要方面不同：最常见的是它们来自一个树突状突起，偶尔包括假定的顶端树突，很少来自核周体的基部;单个细胞可能有2个或更多个这样的突起。较厚的锥形突起（在3 H-尿苷中孵育后）含有大量的RNA;新合成的RNA不会延伸到较细的轴突样突起中。
The growth of the processes of dissociated hippocampal neurons from 18-20 day old rat fetuses was studied in vitro using a method that permits cells to be maintained at relatively low plating densities for periods up to 3 wk (Banker and Cown, 1977). Most of the cells are spherical or ovoid when they first attach to the substratum (polylysine or collagen) but within 24 to 48 h they begin to put out processes. By the end of the 1st wk in culture a significant proportion (.apprx. 45%) come to resemble normal pyramidal cells with a more or less triangular-shaped soma, a single dominant dendrite-like process emerging from the apex of the soma, and several basal dendrites arising from the opposite pole of the cell. Comparisons of the lengths of these dendrite-like processes with those of hippocampal cells in the brains of animals sacrificed on the 4th postnatal day and impregnated by a variant of the Golgi-Cox method indicate that in some cases the rate of process formation in vitro approximates that in vivo and that the general form of the neurons is remarkably like that of immature pyramidal cells. After 1 wk in culture a 2nd type of process can be recognized. These tend to be finer than those identified as dendrites; they are relatively uniform in diameter, frequently give off branches at right angles and in the EM can be seen to form synapses on the larger processes and cell somata. These axon-like processes differ from the axons of normal pyramidal cells in 2 important respects: most commonly they arise from one of the dendritic processes, occasionally including the putative apical dendrites and only rarely from the base of the perikaryon; there may be 2 or more such processes from a single cell. The thicker, tapering processes can be shown (after incubation in 3H-uridine) to contain large amounts of RNA; newly-synthesized RNA does not extend into the finer, axon-like processes.