JIP1基因敲除对青年小鼠视网膜形态和突触蛋白表达的影响
|
摘要
目的探讨JIP1基因对青年小鼠视网膜结构和突触蛋白表达的影响。方法选择1月龄JIP1基因敲除(JIP1-KO)雄鼠和野生型C57雄鼠各12只,通过HE染色观察1月龄野生型C57小鼠和JIP1-KO小鼠视网膜厚度变化,采用Western blot和免疫组化方法检测野生型C57小鼠和JIP1-KO小鼠视网膜神经节细胞标志物(neuronal classⅢβ-Tubulin,TUJ1)、神经丝蛋白(neurofilament, NF)和突触后密度蛋白95(postsynaptic density protein 95, PSD95)、突触素(synaptophysin, SYN)表达。结果 1月龄野生型C57小鼠视网膜全层、外核层(outer nuclear layer,ONL)、内核层(inner nuclear layer,INL)厚度分别为(119.60±11.70)、(41.17±3.59)、(21.90±2.27)μm,JIP1-KO小鼠视网膜全层、ONL层、INL层厚度分别为(110.70±7.21)、(36.83±2.26)、(20.15±1.89)μm,各对应层之间厚度比较均无统计学差异(P>0.05);与野生型C57小鼠相比,JIP1-KO小鼠视网膜中TUJ1、PSD95、SYN表达水平均无统计学差异(P>0.05),TUJ1定位于视网膜神经节细胞层,神经细胞突起NF、PSD95、SYN定位于外网层。结论 JIP1基因敲除对1月龄小鼠视网膜发育影响不显著。
Objective To investigate the role of JNK interacting protein 1(JIP1) gene in retinal structure and synaptic protein expression in young mice. Methods Twelve JIP1 knockout(KO) mice(1 month old) were employed in this study. HE staining was used to observe the changes of retinal thickness in 1-month-old wild type C57 mice and JIP1-KO mice. Western blotting and immunohistochemical assay were used to detect the expression of retinal ganglion cells' markers neuronal class Ⅲ β-tubulin(TUJ1), neurofilament(NF), postsynaptic density protein 95(PSD95) and synaptophysin(SYN) in the 2 groups. Results The thicknesses of the whole retinal layer, the inner nuclear layer(INL) and the outer nuclear layer(ONL) of 1 month old wild type C57 mice were 119.60±11.70, 41.17±3.59 and 21.90±2.27 μm, respectively. And the thicknesses of the 3 layers of the JIP1-KO mice were 110.70±7.21, 36.83±2.26 and 20.15±1.89 μm, respectively. There were no statistical differences in the thickness of the corresponding layers between the 2 kinds of mice(P>0.05). Compared with the wild C57 mice, the expression levels of TUJ1, PSD95 and SYN in the retina of JIP1-KO mice had no significant difference(P>0.05). TUJ1 was located in the retinal ganglion cell layer, while NF, PSD95 and SYN were located in the outer plexiform layer(OPL). Conclusion JIP1 knockout shows no obvious effect on the development of the retina in mice aged 1 month.
Objective To investigate the role of JNK interacting protein 1(JIP1) gene in retinal structure and synaptic protein expression in young mice. Methods Twelve JIP1 knockout(KO) mice(1 month old) were employed in this study. HE staining was used to observe the changes of retinal thickness in 1-month-old wild type C57 mice and JIP1-KO mice. Western blotting and immunohistochemical assay were used to detect the expression of retinal ganglion cells' markers neuronal class Ⅲ β-tubulin(TUJ1), neurofilament(NF), postsynaptic density protein 95(PSD95) and synaptophysin(SYN) in the 2 groups. Results The thicknesses of the whole retinal layer, the inner nuclear layer(INL) and the outer nuclear layer(ONL) of 1 month old wild type C57 mice were 119.60±11.70, 41.17±3.59 and 21.90±2.27 μm, respectively. And the thicknesses of the 3 layers of the JIP1-KO mice were 110.70±7.21, 36.83±2.26 and 20.15±1.89 μm, respectively. There were no statistical differences in the thickness of the corresponding layers between the 2 kinds of mice(P>0.05). Compared with the wild C57 mice, the expression levels of TUJ1, PSD95 and SYN in the retina of JIP1-KO mice had no significant difference(P>0.05). TUJ1 was located in the retinal ganglion cell layer, while NF, PSD95 and SYN were located in the outer plexiform layer(OPL). Conclusion JIP1 knockout shows no obvious effect on the development of the retina in mice aged 1 month.
引文
[1] KLINEDINST S, WANG X, XIONG X, et al. Independent pathways downstream of the Wnd/DLK MAPKKK regulate synaptic structure, axonal transport, and injury signaling[J]. J Neurosci, 2013, 33(31): 12764-12778. DOI:10.1523/JNEUROSCI.5160-12.2013.
[2] ZEKE A, MISHEVA M, REMENYI A, et al. JNK signaling: regulation and functions based on complex protein-protein partnerships[J]. Microbiol Mol Biol Rev, 2016, 80(3): 793-835. DOI: 10.1128/MMBR.00043-14.
[3] MOREL C, SHERRIN T, KENNEDDY N J, et al. JIP1-mediated JNK activation negatively regulates synaptic plasticity and spatial memory[J]. J Neurosci, 2018, 37(11): 3708-3728.DOI:10.1523/JNEUROSCI.1913-17.2018.
[4] CHIBA K, CHIEN K Y, SOBU Y, et al. Phosphorylation of KLC1 modifies interaction with JIP1 and abolishes the enhanced fast velocity of APP transport by kinesin-1[J]. Mol Biol Cell, 2017, 28(26): 3857-3869. DOI:10.1091/mbc.E17-05-0303.
[5] SUM T, LI Y, LI T, et al. JIP1 and JIP3 cooperate to mediate TrkB anterograde axonal transport by activating kinesin-1[J]. Cell Mol Life Sci, 2017, 74(21): 4027-4044. DOI: 10.1007/s00018-017-2568-z.
[6] DENG C Y, LEI W L, XU X H, et al. JIP1 mediates anterograde transport of Rab10 cargos during neuronal polarization[J]. J Neurosci, 34(5): 1710-1723. DOI: 10.1523/JNEUROSCI.4496-13.2014.
[7] JUNWON L, SANG H C, YOUNG B K, et al. Defined conditions for differentiation of functional retinal ganglion cells from human pluripotent stem cells[J]. IOVS, 2018, 59(8): 3531-3545. DOI: 10.1167/iovs.17-23439.
[8] MARGEVICIUS D R, BASTIAN C, FAN Q, et al. JNK-interacting protein 1 mediates Alzheimer’s-like pathological features in AICD-transgenic mice[J]. Neurobiol Aging, 2015, 36(8): 2370-2379. DOI: 10.1016/j.neurobiolaging.2015.04.013.
[9] DAJASBAILADOR F, BANTOUNAS I, JONES E V, et al. Regulation of axon growth by the JIP1-AKT axis[J]. J Cell Sci, 2014, 127(Pt 1): 230-239. DOI: 10.1242/jcs.137208.
[10] WANG P, ZHANG J Y, RICHARDSON G P, et al. Two cytokine signaling molecules co-operate to promote axonal transport and growth[J]. Exp Neurol, 2011, 228(2): 165-172. DOI: 10.1016/j.expneurol.2010.08.021.
[11] BARNAT M, ENSLEN H, PROPST F,et al. Distinct roles of c-Jun N-terminal kinase isoforms in neurite initiation and elongation during axonal regeneration[J]. J Neurosci, 2010, 30(23): 7804-7816.DOI:10.1523/JNEUROSCI.0372-10.2010.
[12] DAJAS-BAILAFOR F, JONES E V, WHITMARSH A J. The JIP1 scaffold protein regulates axonal development in cortical neurons[J]. Curr Biol, 2008, 18(3): 221-226. DOI: 10.1016/j.cub.2008.01.025.
[13] HEIN T W, WEN X, XIN X, et al. Acute and chronic hyperglycemia elicit JIP1/JNK-mediated endothelial vasodilator dysfunction of retinal arterioles[J]. IOVS, 2016, 57(10): 4333-4343. DOI: 10.1167/iovs.16-19990.
[14] ZHANG Y, QIU B, WANG J, et al. Effects of BDNF-transfected BMSCs on neural functional recovery and synaptophysin expression in rats with cerebral infarction[J]. Mol Neurobiol, 2017, 54(5): 1-12. DOI: 10.1007/s12035-016-9946-7.
[15] LING Y Z, MA W, YU L, et al. Decreased PSD95 expression in medial prefrontal cortex (mPFC) was associated with cognitive impairment induced by sevoflurane anesthesia[J]. J Zhejiang Univ Sci B, 2015,16(9): 763-771. DOI: 10.1631/jzus.B1500006.
[16] HAITAO W, MINFEI W, CHUANJUN Z, et al. Neurofilament proteins in axonal regeneration and neurodegenerative diseases[J]. Neural Regen Res, 2012, 7(8): 620-626. DOI: 10.3969/j.issn.1673-5374.2012.08.010.
[17] BRAUNGER B M, DEMMER C, TAMM E R. Oxygen Transport to Tissue XXXIII [M]. New York: Springer Science Business Media, 2014: 9-13.
[2] ZEKE A, MISHEVA M, REMENYI A, et al. JNK signaling: regulation and functions based on complex protein-protein partnerships[J]. Microbiol Mol Biol Rev, 2016, 80(3): 793-835. DOI: 10.1128/MMBR.00043-14.
[3] MOREL C, SHERRIN T, KENNEDDY N J, et al. JIP1-mediated JNK activation negatively regulates synaptic plasticity and spatial memory[J]. J Neurosci, 2018, 37(11): 3708-3728.DOI:10.1523/JNEUROSCI.1913-17.2018.
[4] CHIBA K, CHIEN K Y, SOBU Y, et al. Phosphorylation of KLC1 modifies interaction with JIP1 and abolishes the enhanced fast velocity of APP transport by kinesin-1[J]. Mol Biol Cell, 2017, 28(26): 3857-3869. DOI:10.1091/mbc.E17-05-0303.
[5] SUM T, LI Y, LI T, et al. JIP1 and JIP3 cooperate to mediate TrkB anterograde axonal transport by activating kinesin-1[J]. Cell Mol Life Sci, 2017, 74(21): 4027-4044. DOI: 10.1007/s00018-017-2568-z.
[6] DENG C Y, LEI W L, XU X H, et al. JIP1 mediates anterograde transport of Rab10 cargos during neuronal polarization[J]. J Neurosci, 34(5): 1710-1723. DOI: 10.1523/JNEUROSCI.4496-13.2014.
[7] JUNWON L, SANG H C, YOUNG B K, et al. Defined conditions for differentiation of functional retinal ganglion cells from human pluripotent stem cells[J]. IOVS, 2018, 59(8): 3531-3545. DOI: 10.1167/iovs.17-23439.
[8] MARGEVICIUS D R, BASTIAN C, FAN Q, et al. JNK-interacting protein 1 mediates Alzheimer’s-like pathological features in AICD-transgenic mice[J]. Neurobiol Aging, 2015, 36(8): 2370-2379. DOI: 10.1016/j.neurobiolaging.2015.04.013.
[9] DAJASBAILADOR F, BANTOUNAS I, JONES E V, et al. Regulation of axon growth by the JIP1-AKT axis[J]. J Cell Sci, 2014, 127(Pt 1): 230-239. DOI: 10.1242/jcs.137208.
[10] WANG P, ZHANG J Y, RICHARDSON G P, et al. Two cytokine signaling molecules co-operate to promote axonal transport and growth[J]. Exp Neurol, 2011, 228(2): 165-172. DOI: 10.1016/j.expneurol.2010.08.021.
[11] BARNAT M, ENSLEN H, PROPST F,et al. Distinct roles of c-Jun N-terminal kinase isoforms in neurite initiation and elongation during axonal regeneration[J]. J Neurosci, 2010, 30(23): 7804-7816.DOI:10.1523/JNEUROSCI.0372-10.2010.
[12] DAJAS-BAILAFOR F, JONES E V, WHITMARSH A J. The JIP1 scaffold protein regulates axonal development in cortical neurons[J]. Curr Biol, 2008, 18(3): 221-226. DOI: 10.1016/j.cub.2008.01.025.
[13] HEIN T W, WEN X, XIN X, et al. Acute and chronic hyperglycemia elicit JIP1/JNK-mediated endothelial vasodilator dysfunction of retinal arterioles[J]. IOVS, 2016, 57(10): 4333-4343. DOI: 10.1167/iovs.16-19990.
[14] ZHANG Y, QIU B, WANG J, et al. Effects of BDNF-transfected BMSCs on neural functional recovery and synaptophysin expression in rats with cerebral infarction[J]. Mol Neurobiol, 2017, 54(5): 1-12. DOI: 10.1007/s12035-016-9946-7.
[15] LING Y Z, MA W, YU L, et al. Decreased PSD95 expression in medial prefrontal cortex (mPFC) was associated with cognitive impairment induced by sevoflurane anesthesia[J]. J Zhejiang Univ Sci B, 2015,16(9): 763-771. DOI: 10.1631/jzus.B1500006.
[16] HAITAO W, MINFEI W, CHUANJUN Z, et al. Neurofilament proteins in axonal regeneration and neurodegenerative diseases[J]. Neural Regen Res, 2012, 7(8): 620-626. DOI: 10.3969/j.issn.1673-5374.2012.08.010.
[17] BRAUNGER B M, DEMMER C, TAMM E R. Oxygen Transport to Tissue XXXIII [M]. New York: Springer Science Business Media, 2014: 9-13.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |