Precise control of ion channel and gap junction expression is required for patterning of the regenerating axolotl limb.
Precise control of ion channel and gap junction expression is required for patterning of the regenerating axolotl limb.
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DOI：
10.1387/ijdb.200114jw
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Whited JL
中科院分区：
文献类型：
--
作者：
Sousounis K;Erdogan B;Levin M;Whited JL
蝾螈和其他蝾螈有能力在截肢或受伤后再生失去的组织。在许多情况下，细胞群内的生长和形态发生是通过转录网络和生物物理特性（如离子流和电压梯度）的相互作用来协调的。然而，目前尚不清楚细胞离子状态的调节是否与再生后期的肢体模式有关。本研究在蝾螈肢胚细胞中调控离子通道和间隙连接的表达和活性。在逆转录病毒感染后截肢，以驱动人类间隙连接蛋白Connexin 26 （Cx26）、钾（Kir2.1-Y242F和Kv1.5）和钠（NeoNav1.5）离子通道蛋白的表达以及EGFP控制。骨骼制备显示，与表达EGFP的对照肢相比，过表达Cx26导致并指，而过表达离子通道蛋白导致手指缺失和结构异常。此外，我们发现在再生过程中，将四肢暴露于间隙连接抑制剂林丹会导致手指丢失。我们的数据表明，操纵原生离子通道和缝隙连接功能会导致再生指骨数量和结构的图案缺陷。间隙连接和离子通道已被证明可以介导离子流动，控制内源性电压梯度，而内源性电压梯度与基因表达、细胞周期进程、迁移和其他细胞行为的调节密切相关。因此，我们假设这些通道的错误表达可能扰乱了这种调节，导致细胞行为不协调，从而导致形态缺陷。
Axolotls and other salamanders have the capacity to regenerate lost tissue after an amputation or injury. Growth and morphogenesis are coordinated within cell groups in many contexts by the interplay of transcriptional networks and biophysical properties such as ion flows and voltage gradients. It is not, however, known whether regulators of a cell’s ionic state are involved in limb patterning at later stages of regeneration. Here we manipulated expression and activities of ion channels and gap junctions in vivo, in axolotl limb blastema cells. Limb amputations followed by retroviral infections were performed to drive expression of a human gap junction protein Connexin 26 (Cx26), potassium (Kir2.1-Y242F and Kv1.5) and sodium (NeoNav1.5) ion channel proteins along with EGFP control. Skeletal preparation revealed that overexpressing Cx26 caused syndactyly, while overexpression of ion channel proteins resulted in digit loss and structural abnormalities compared to EGFP expressing control limbs. Additionally, we showed that exposing limbs to the gap junction inhibitor lindane during the regeneration process caused digit loss. Our data reveal that manipulating native ion channel and gap junction function in blastema cells results in patterning defects involving the number and structure of the regenerated digits. Gap junctions and ion channels have been shown to mediate ion flows that control the endogenous voltage gradients which are tightly associated with the regulation of gene expression, cell cycle progression, migration, and other cellular behaviors. Therefore, we postulate that mis-expression of these channels may have disturbed this regulation causing uncoordinated cell behavior which results in morphological defects.
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影响因子：
3.8
作者：
GUAN, XJ;BONNEY, WJ;RUCH, RJ
通讯作者：
RUCH, RJ
影响因子：
5.7
作者：
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影响因子：
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作者：
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内源性电场是用于细胞迁移的指导提示。
影响因子：
4
作者：
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通讯作者：
Funk RH
影响因子：
3.4
作者：
Durant, Fallon;Bischof, Johanna;Levin, Michael
通讯作者：
Levin, Michael