Distinct axo-protective and axo-destructive roles for Schwann cells after injury in a novel compartmentalised mouse myelinating coculture system
Distinct axo-protective and axo-destructive roles for Schwann cells after injury in a novel compartmentalised mouse myelinating coculture system
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在新型小鼠髓鞘共培养系统中损伤后雪旺细胞具有独特的轴突保护和轴突破坏作用
DOI：
10.1101/2023.05.19.541371
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Mutschler C
中科院分区：
文献类型：
--
作者：
Mutschler C
在过去的40年里，雪旺细胞(SC) -背根神经节（DRG）神经元共培养已经成为了解周围神经系统（PNS）髓鞘形成、损伤和再生过程中细胞-细胞信号传导和相互作用的重要技术。虽然使用大鼠SCs和大鼠DRG神经元的方法是常见的，但在该领域还没有建立的方案来描述在游离髓鞘共培养中使用小鼠SCs和小鼠DRG神经元。有一个非常需要这样一个协议，因为这将允许使用来自许多不同的转基因小鼠系的细胞。在这里，我们描述了一种方案，共同培养分离的小鼠SCs和DRG神经元，并诱导强大的髓鞘形成。使用微流控室可以进行药物治疗的流体分离，可以对培养物进行解剖以研究损伤反应，并且可以很容易地用慢病毒转染细胞以进行实时成像。我们使用这个模型来量化创伤性轴突切开术后，髓鞘化SCs的存在和缺失以及未诱导髓鞘化的轴突排列SCs的退化率。我们发现，无论髓鞘形成状态如何，SCs都具有轴突保护作用，并在早期延缓轴突退化。在损伤后的后期时间点，我们使用共培养的实时成像显示，一旦轴突变性开始，SCs就会分解、摄取并清除轴突碎片。摘要：一种新的区隔化解离小鼠髓鞘SC-DRG共培养系统揭示了创伤后雪旺细胞对轴突完整性的不同轴保护和轴破坏阶段。
Myelinating Schwann cell (SC)– dorsal root ganglion (DRG) neuron cocultures have been an important technique over the last four decades in understanding cell-cell signalling and interactions during peripheral nervous system (PNS) myelination, injury, and regeneration. While methods using rat SCs and rat DRG neurons are commonplace, there are no established protocols in the field describing the use of mouse SCs with mouse DRG neurons in dissociated myelinating cocultures. There is a great need for such a protocol as this would allow the use of cells from many different transgenic mouse lines. Here we describe a protocol to coculture dissociated mouse SCs and DRG neurons and induce robust myelination. Use of microfluidic chambers permits fluidic isolation for drug treatments, allows cultures to be axotomised to study injury responses, and cells can readily be transfected with lentiviruses to permit live imaging. We used this model to quantify the rate of degeneration after traumatic axotomy in the presence and absence of myelinating SCs and axon aligned SCs that were not induced to myelinate. We find that SCs, irrespective of myelination status, are axo-protective and delay axon degeneration early on. At later time points after injury, we use live imaging of cocultures to show that once axonal degeneration has commenced SCs break up, ingest, and clear axonal debris.Summary statementA novel compartmentalised dissociated mouse myelinating SC-DRG coculture system reveals distinct axo-protective and axo-destructive phases of Schwann cells on axon integrity after trauma.
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影响因子：
7.2
作者：
Salzer, James L.
通讯作者：
Salzer, James L.
颈上神经节外植体培养中的轴突变性测定。
影响因子：
--
作者：
Andrea Loreto;J. Gilley
通讯作者：
J. Gilley
影响因子：
13.6
作者：
Hyung S;Lee SR;Kim J;Kim Y;Kim S;Kim HN;Jeon NL
通讯作者：
Jeon NL
使用微流体室模拟 PNS 和 CNS 髓鞘形成。
影响因子：
--
作者：
Adrien Vaquié;Alizée Sauvain;C. Jacob
通讯作者：
C. Jacob
影响因子：
5.3
作者：
Rosenberg, Allison F.;Isaacman-Beck, Jesse;Granato, Michael
通讯作者：
Granato, Michael