Helke C J, Verdier-Pinard D
Department of Pharmacology and Neuroscience Program, Uniformed Services University of the Health Science, 4301 Jones Bridge Road, Bethesda, MD 20814-4799, USA.
Brain Res. 2000 Nov 24;884(1--2):206-12. doi: 10.1016/s0006-8993(00)02988-7.
Mature nodose and petrosal ganglia neurons (placodally derived afferent neurons of the vagal and glossopharyngeal nerves) contain TrkA and TrkC, and transport specific neurotrophins [nerve growth factor (NGF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4)]. This study evaluated neurotrophin influences on the presence of neuropeptides and/or neurotransmitter enzymes in these visceral sensory neurons. NGF, NT-3 and NT-4 (10-100 ng/ml) were applied (5 days) to dissociated, enriched, cultures of mature nodose/petrosal ganglia neurons, and the neurons processed for tyrosine hydroxylase (TH), vasoactive intestinal peptide (VIP), calcitonin gene-related peptide (CGRP) and neurofilament (NF-200) immunocytochemistry. Addition of NGF to nodose/petrosal ganglia neuron-enriched cultures significantly increased the number of TH-immunoreactive (ir) neurons, decreased the number of VIP-ir neurons in the cultures, and did not affect the numbers of CGRP-ir neurons. The addition of an NGF neutralizing antibody attenuated the effects of NGF on TH and VIP-ir neurons. NT-3 increased the number of VIP-ir neurons in the nodose/petrosal ganglia cultures and did not alter the numbers of TH-, or CGRP-ir neurons. The addition of an NT-3 neutralizing antibody attenuated the effects of NT-3 on VIP-ir neurons. NT-4 had no significant effects on the numbers of TH, VIP and CGRP-ir neurons. The absence of neurotrophin-induced changes in the numbers of NF-200-ir neurons in culture showed the lack of neurotrophin-mediated changes in survival of mature vagal afferent neurons. These data demonstrate that specific neurotrophins influence the numbers of neurons labeled for specific neurochemicals in nodose/petrosal ganglia cultures. These data, coupled with previous evidence for the presence of TrkA and TrkC mRNA and of the retrograde transport of NGF and NT-3, suggest important roles for NGF and NT-3 in the maintenance of transmitter phenotype of these mature visceral afferent neurons.
成熟的结状神经节和岩神经节神经元(迷走神经和舌咽神经的板层衍生传入神经元)含有TrkA和TrkC,并运输特定的神经营养因子[神经生长因子(NGF)、神经营养因子-3(NT-3)、神经营养因子-4(NT-4)]。本研究评估了神经营养因子对这些内脏感觉神经元中神经肽和/或神经递质酶存在的影响。将NGF、NT-3和NT-4(10 - 100 ng/ml)应用于解离、富集的成熟结状/岩神经节神经元培养物(5天),并对神经元进行酪氨酸羟化酶(TH)、血管活性肠肽(VIP)、降钙素基因相关肽(CGRP)和神经丝(NF-200)免疫细胞化学处理。向富含结状/岩神经节神经元的培养物中添加NGF显著增加了TH免疫反应性(ir)神经元的数量,减少了培养物中VIP-ir神经元的数量,并且不影响CGRP-ir神经元的数量。添加NGF中和抗体减弱了NGF对TH和VIP-ir神经元的作用。NT-3增加了结状/岩神经节培养物中VIP-ir神经元的数量,并且不改变TH-ir或CGRP-ir神经元的数量。添加NT-3中和抗体减弱了NT-3对VIP-ir神经元的作用。NT-4对TH、VIP和CGRP-ir神经元的数量没有显著影响。培养物中神经营养因子诱导的NF-200-ir神经元数量变化的缺失表明成熟迷走传入神经元的存活缺乏神经营养因子介导的变化。这些数据表明特定的神经营养因子影响结状/岩神经节培养物中标记特定神经化学物质的神经元数量。这些数据,再加上先前关于TrkA和TrkC mRNA存在以及NGF和NT-3逆行运输的证据,表明NGF和NT-3在维持这些成熟内脏传入神经元的递质表型方面起着重要作用。