Zafirov S, Heimrich B, Frotscher M
Institute of Anatomy, University of Freiburg, Germany.
J Comp Neurol. 1994 Jul 15;345(3):472-80. doi: 10.1002/cne.903450312.
Dendrites and spines are postsynaptic structures that develop in association with presynaptic fibers. Recent studies have shown that granule cells of the fascia dentata survive in slice cultures and differentiate in a manner known from in situ studies. However, all extrinsic afferent fibers are absent under culture conditions. In the present study, we study whether dendrites and spines of granule cells in slice cultures differentiate normally, although they are not contacted by their normal layer-specific afferents. Slices of hippocampus were prepared from rat pups at the day of birth. After 5, 10, 15, and 20 days of incubation, granule cells in these cultures were Golgi impregnated. For comparison, perfusion-fixed hippocampal sections of 5-, 10-, 15-, and 20-day-old rats were impregnated the same way. Our results show that the total density of spines on granule cell dendrites in culture increased as in perfusion-fixed animals. However, after 20 days of incubation, the absolute number of dendritic spines on cultured neurons was reduced because of a reduction of peripheral dendrites. This reduction was accompanied by an increase in the number of stem dendrites originating from the perikaryon. The density of spines on these proximal dendrites was larger in cultured granule cells than in controls. Our results suggest that the lack of major extrinsic (entorhinal) afferents that normally terminate on peripheral granule cell dendrites causes retraction of these dendrites. At the same time, there is growth of proximal dendritic portions. Proximal dendrites are targets of associational fibers, which are known to sprout under these culture conditions.
树突和棘是与突触前纤维相关发育的突触后结构。最近的研究表明,齿状回颗粒细胞在脑片培养物中存活,并以原位研究中已知的方式分化。然而,在培养条件下所有外在传入纤维均不存在。在本研究中,我们研究脑片培养物中颗粒细胞的树突和棘是否能正常分化,尽管它们未与正常的层特异性传入纤维接触。出生当天从新生大鼠幼崽制备海马脑片。培养5、10、15和20天后,对这些培养物中的颗粒细胞进行高尔基染色。作为对照,对5、10、15和20日龄大鼠的灌注固定海马切片进行同样的染色。我们的结果表明,培养物中颗粒细胞树突上棘的总密度与灌注固定动物中的情况一样增加。然而,培养20天后,由于外周树突减少,培养神经元上树突棘的绝对数量减少。这种减少伴随着源自胞体的主干树突数量增加。培养的颗粒细胞中这些近端树突上棘的密度比对照组大。我们的结果表明,通常终止于外周颗粒细胞树突上的主要外在(内嗅)传入纤维的缺失导致这些树突回缩。与此同时,近端树突部分生长。近端树突是联合纤维的靶点,已知在这些培养条件下联合纤维会出芽。