Wentzek L A, Bowers C W, Khairallah L, Pilar G
Physiology and Neurobiology Department, University of Connecticut, Storrs 06269-3042.
J Neurosci. 1993 Jul;13(7):3143-54. doi: 10.1523/JNEUROSCI.13-07-03143.1993.
It is well established that during in vivo development the neurons of the avian ciliary ganglion are dependent for their survival on structures in the eye. Separate neuron populations innervate intraocular smooth and striated muscle targets. All ciliary neurons survive when cocultured with striated muscle. We demonstrate that when ciliary ganglion neurons are plated on explants of the choroid coat (a smooth muscle-containing target tissue) using a defined medium (N2), the neurons survive and grow vigorously into the tissue, forming contacts between axons and target cells identified as smooth muscle. Conditioned medium from choroid explants also rescues all the neurons, as does coculturing ciliary ganglion neurons with dissociated choroid cells. However, the presence of horse serum and chick embryo extract in the medium inhibits the choroid's ability to support ciliary neurons. The effects of these additives on the phenotypic expression of the smooth muscle may explain the inability of previous investigators to demonstrate target-derived support from smooth muscle preparations. Because the choroid contains cell types other than smooth muscle (e.g., fibroblasts and endothelial cells), we could not identify smooth muscle as the only cell type responsible for the release of the soluble trophic factor present in the target tissue. However, indirect evidence using avian primary fibroblast cultures, a fibroblast cell line, and an anatomically simple smooth muscle preparation, the avian amnion, suggests that smooth muscle cells are sufficient to account for the observed trophic activity, and that similar target-derived molecules support the survival of both types of ciliary ganglion cells.
众所周知,在体内发育过程中,鸟类睫状神经节的神经元依靠眼睛中的结构来维持其存活。不同的神经元群体支配眼内的平滑肌和横纹肌靶标。当与横纹肌共培养时,所有睫状神经元都能存活。我们证明,当使用限定培养基(N2)将睫状神经节神经元接种到脉络膜外植体(一种含平滑肌的靶组织)上时,神经元能够存活并有力地生长到该组织中,在轴突与被鉴定为平滑肌的靶细胞之间形成联系。来自脉络膜外植体的条件培养基也能拯救所有神经元,将睫状神经节神经元与解离的脉络膜细胞共培养也有同样效果。然而,培养基中马血清和鸡胚提取物的存在会抑制脉络膜支持睫状神经元的能力。这些添加剂对平滑肌表型表达的影响可能解释了先前研究人员无法证明平滑肌制剂提供靶源性支持的原因。由于脉络膜除了平滑肌外还包含其他细胞类型(如成纤维细胞和内皮细胞),我们无法确定平滑肌是靶组织中释放可溶性营养因子的唯一细胞类型。然而,使用禽类原代成纤维细胞培养物、一种成纤维细胞系以及一种解剖结构简单的平滑肌制剂——禽类羊膜的间接证据表明,平滑肌细胞足以解释所观察到的营养活性,并且类似的靶源性分子支持两种类型的睫状神经节细胞的存活。