Survival and morphology of auditory neurons in dissociated cultures of newborn mouse spiral ganglion
Survival and morphology of auditory neurons in dissociated cultures of newborn mouse spiral ganglion
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DOI：
10.1016/j.neuroscience.2005.11.030
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发表时间：
2006-01-01
期刊：
影响因子：
3.3
通讯作者：
Richter, CP
中科院分区：
文献类型：
--
作者：
Whitlon, DS;Ketels, KV;Richter, CP
我们系统地描述了新生/出生后1天小鼠耳蜗培养物中神经元的存活和生长。耳蜗螺旋神经节的分离培养为研究听觉神经元的存活、发育和生理的分子机制提供了实验环境。为了将存活率与源组织中存在的神经元总数联系起来，我们将来自不同CD-1新生小鼠的3只耳蜗包埋在Araldite树脂中，并以5 μ m的厚度连续切片。计数所有神经元。为了避免重复计数，每个部分作为下一个查找部分，每个神经节给出8240 +/- 423 （sd）神经元。维持在邻近非神经组织、脑源性神经营养因子、神经营养因子3、白血病抑制因子（LIF）和10%胎牛血清存在下的培养在镀后42小时恢复了最佳的总存活率（30%）。最佳总生存率要求血清成分持续存在大于100,000 MW。在有或没有LIF的情况下，镀效率（4小时后附着在孔上的神经元数量）相似。在42小时的培养过程中，LIF的加入使神经元的存活率达到100%；没有LIF，很大一部分神经元无法存活。LIF似乎通过优先保存双极神经元群体来维持生存，而对单极神经元的数量几乎没有影响。这项工作提供了体外听觉神经元存活和形态的定量测量。这些结果支持了螺旋神经节神经元在体内的存活可能依赖于与邻近的非神经组织的相互作用的观点，并提出了毛细胞损伤后双极形态的维持可能需要生化机制，而不是神经营养素诱导的机制。(c) 2005年由Elsevier Ltd代表IBRO出版。
We have systematically characterized neuronal survival and growth in cultures derived from newborn/postnatal day I mouse cochlea. Dissociated cultures of the cochlear spiral ganglion provide an experimental environment in which to examine molecular mechanisms of survival, development and physiology of auditory neurons. To relate survival to the total number of neurons present in the source tissue, three cochleas from different newborn CD-1 mice were embedded in Araldite resin and serially sectioned at 5 mu m thickness. All neurons were counted. To avoid overcounting, each section served as a lookup section for the next, giving 8240 +/- 423 (S.D.) neurons per ganglion. Cultures maintained in the presence of adjacent non-neural tissue, brain-derived neurotrophic factor, neurotrophin 3, leukemia inhibitory factor (LIF) and 10% fetal bovine serum returned the best overall survival (30%) at 42 h post-plating. Best overall survival required the continuous presence of a serum component(s) larger than 100,000 MW. Plating efficiency (number of neurons that attach to the well after 4 h) was similar in the presence or absence of LIF. Inclusion of LIF maintained 100% survival of plated neurons over 42 h of culture; without LIF, a large fraction of the neurons did not survive. LIF appeared to maintain survival by preferentially preserving a population of bipolar neurons, while having little effect on the number of monopolar neurons. This work provides quantitative measures of survival and morphology of auditory neurons in vitro. The results support the idea that survival of spiral ganglion neurons in vivo may depend on interactions with adjacent, non-neural tissue and raise the possibility that maintenance of bipolar morphology after hair cell damage may require biochemical mechanisms in addition to those induced by neurotrophins. (c) 2005 Published by Elsevier Ltd on behalf of IBRO.