Sanes D H, Hafidi A
Center for Neural Science, New York University, New York 10003, USA.
J Neurobiol. 1996 Dec;31(4):503-11. doi: 10.1002/(SICI)1097-4695(199612)31:4<503::AID-NEU9>3.0.CO;2-D.
We previously demonstrated that inhibitory synaptic transmission influences dendrite development in vivo. We now report an analogous finding in an organotypic culture of a glycinergic projection nucleus, the medial nucleus of the trapezoid body (MNTB), and its postsynaptic target, the lateral superior olive (LSO) of gerbils. Cultures were generated at 6-7 days postnatal and grown in serum containing medium with or without the glycine receptor antagonist, strychnine (SN), at 2 microM. LSO neurons were then labeled with biocytin, and the dendritic arbors were analyzed morphometrically. Compared to neurons form age-matched in vivo tissue, the neurons cultured in control media were somewhat atrophic, including decreases in dendritic branching and length. Incubation in strychnine led to a dramatic increase in dendritic branching and total dendritic length. Control neurons averaged 6.3 branches compared to 18 branches/neuron in SN-treated cultures. There was a similar increase in primary dendrites and total dendritic length. The physical elimination of MNTB cells did not mimic SN treatment, presumably because glycinergic LSO neurons generated intrinsic connections. In fact, the LSO soma area was significantly greater following MNTB removal, suggesting that these afferents provide a second signal to postsynaptic neurons. These results suggest that spontaneous glycinergic transmission regulates the growth of postsynaptic processes.
我们先前证明,抑制性突触传递在体内影响树突发育。我们现在报告在沙鼠甘氨酸能投射核——梯形体内侧核(MNTB)及其突触后靶标——外侧上橄榄核(LSO)的器官型培养物中的类似发现。在出生后6 - 7天制备培养物,并在含有2 microM甘氨酸受体拮抗剂士的宁(SN)的血清培养基中生长,或不添加该拮抗剂。然后用生物素标记LSO神经元,并对树突分支进行形态计量学分析。与来自年龄匹配的体内组织的神经元相比,在对照培养基中培养的神经元有些萎缩,包括树突分支和长度的减少。在士的宁中孵育导致树突分支和总树突长度显著增加。对照神经元平均有6.3个分支,而在SN处理的培养物中每个神经元平均有18个分支。初级树突和总树突长度也有类似增加。物理去除MNTB细胞并不能模拟SN处理,推测是因为甘氨酸能LSO神经元产生了内在连接。事实上,去除MNTB后LSO胞体面积显著增大,表明这些传入神经为突触后神经元提供了第二个信号。这些结果表明,自发的甘氨酸能传递调节突触后突起的生长。