Lindsay R M, Rohrer H
Dev Biol. 1985 Nov;112(1):30-48. doi: 10.1016/0012-1606(85)90116-2.
Explant and dissociated neuron-enriched cultures of nodose ganglia (inferior or distal sensory ganglion of the Xth cranial nerve) were established from chick embryos taken between embryonic Day 4 (E4) and Day 16 (E16). The response of each type of culture to nerve growth factor (NGF) was examined over this developmental range. At the earliest ages taken (E4-E6), NGF elicited modest neurite outgrowth from ganglion explants cultured in collagen gel for 24 hr, although the effect of NGF on ganglia taken from E4 chicks was only marginally greater than spontaneous neurite extension from control ganglia of the same developmental age. The response of nodose explants to NGF was maximal at E6-E7, but declined to a negligible level in ganglia taken from E9-E10 or older chick embryos. In dissociated neuron-enriched cultures, nodose ganglion neurons were unresponsive to NGF throughtout the entire developmental age range between E5 and E12. In contrast to the lack of effect of NGF, up to 50% of nodose ganglion neurons survived and produced extensive neurites in dissociated cultures, on either collagen- or polylysine-coated substrates, in the presence of extracts of late embryonic or early posthatched chick liver (E18-P7). Antiserum to mouse NGF did not block the neurotrophic activity of chick (or rat or bovine) liver extracts. Whether cultured with chick liver extract alone or with chick liver extract plus NGF, nodose ganglion neurons taken from E6-E12 chick embryos and maintained in culture for 2 days were devoid of NGF receptors, as assessed by autoradiography of cultures incubated with 125I-NGF. Under similar conditions 70-95% of spinal sensory neurons (dorsal root ganglion--DRG) were heavily labeled. 2+
从胚胎第4天(E4)至第16天(E16)的鸡胚中建立了结节神经节(第10对脑神经的下或远侧感觉神经节)的外植体和富含解离神经元的培养物。在此发育范围内研究了每种培养物对神经生长因子(NGF)的反应。在最早取材的时期(E4 - E6),NGF可使培养在胶原凝胶中24小时的神经节外植体产生适度的神经突起生长,尽管NGF对取自E4鸡胚的神经节的作用仅略大于相同发育年龄对照神经节的自发神经突起延伸。结节外植体对NGF的反应在E6 - E7时最大,但在取自E9 - E10或更老鸡胚的神经节中降至可忽略不计的水平。在富含解离神经元的培养物中,结节神经节神经元在E5至E12的整个发育年龄范围内对NGF均无反应。与NGF缺乏作用形成对比的是,在晚期胚胎或早期孵出后鸡肝(E18 - P7)提取物存在的情况下,高达50%的结节神经节神经元在解离培养物中存活并产生广泛的神经突起,培养物置于胶原或聚赖氨酸包被的底物上。抗小鼠NGF的抗血清不能阻断鸡(或大鼠或牛)肝提取物的神经营养活性。通过用125I - NGF孵育培养物的放射自显影评估,取自E6 - E12鸡胚并在培养中维持2天的结节神经节神经元,无论单独用鸡肝提取物培养还是与鸡肝提取物加NGF一起培养,均无NGF受体。在类似条件下,70 - 95%的脊髓感觉神经元(背根神经节 - DRG)被大量标记。2+