PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of neuroscibullNeuroscience Bulletin
 
Neurosci Bull. 2007 July; 23(4): 203–208.
Published online 2008 February 1. doi:  10.1007/s12264-007-0030-1
PMCID: PMC5550582

Language: English | Chinese

Expression changes of microtubule associated protein 1B in the brain of Fmr1 knockout mice

微맜相关蛋白 1B 在 Fmr1 基因敲除小鼠脑组织内的表达变化

Abstract

Objective

To explore the regulatory effect of fragile X mental retardation protein (FMRP) on the translation of microtubule associated protein 1B (MAP1B).

Methods

The expressions of MAP1B protein and MAP1B mRNA in the brains of 1-week and 6-week old fragile X mental retardation-1 (Fmr1) knockout (KO) mice were investigated by immunohistochemistry, Western blot, and in situ hybridization, with the age-matched wild type mice (WT) as controls.

Results

The mean optical density (MOD) of MAP1B was significantly decreased in each brain region in KO6W compared with WT6W, whereas in KO1W, this decrease was only found in the hippocampus and cerebellum. MAP1B in 6-week mice was much less than that in 1-week mice of the same genotype. The results of Western blot and in situ hybridization showed that MAP1B protein and MAP1B mRNA were significantly decreased in the hippocampus of both KO1W and KO6W.

Conclusion

The decreased MAP1B protein and MAP1B mRNA in the Fmr1 knockout mice indicate that FMRP may positively regulate the expression of MAP1B.

Keywords: fragile X syndrome, fragile X mental retardation protein, microtubule associated protein 1B, mice

摘要

目的

探讨脆性X智能低下蛋白(fragile X mental retardation protein, FMRP)对微맜相关蛋白1B(microtubule associated protein 1B, MAP1B) 是否具有调控作用。

方法

应用免疫组化、 免疫印记和原位杂交的方法, 对 1 周龄和 6 周龄的 Fmr1 基因敲除型 (KO) 和同龄野生型 (WT) 小鼠脑组织 MAP1B 及 MAP1B mRNA 进行分析。

结果

免疫组化的结果显示: 6 周龄 KO 小鼠各个脑区 MAP1B 的平均光密度值 (MOD) 值均显著低于同龄 WT 小鼠 (P < 0.05), 1 周龄 KO 小鼠仅在小脑和海马显著降低 (P < 0.01); 各脑区 MAP1B 的 MOD 值在 6 周龄小鼠均比同基因型的 1 周龄小鼠显著降低(P < 0.05)。 免疫印记和原位杂交结果分别显示 MAP1B 及 MAP1B mRNA 在 KO 小鼠的海马组织均显著降低(P < 0.05)。

结论

MAP1B和 MAP1B mRNA在 Fmr1 基因敲除小鼠脑组织的表达均显著减少, 提示 FMRP 可能正性调节 MAP1B 的表达。

关键词: 脆性X 综合征, 脆性X 智能低下蛋白, 微맜相关蛋白1B, s小鼠

Reference

[1] Brown V., Jin P., Ceman S., Darnell J.C., O’Donnell W.T., Tenenbaum S.A., et al. Microarray identification of FMRP-associated brain mRNAs and altered mRNA translational profiles in fragile X syndrome. Cell. 2001;107:477–487. doi: 10.1016/S0092-8674(01)00568-2. [PubMed] [Cross Ref]
[2] Darnell J.C., Jensen K.B., Jin P., Brown V., Warren S.T., Darnell R.B. Fragile X mental retardation protein targets G quartet mRNAs important for neuronal function. Cell. 2001;107:489–499. doi: 10.1016/S0092-8674(01)00566-9. [PubMed] [Cross Ref]
[3] Miyashiro K.Y., Beckel-Mitchener A., Purk T.P., Becker K.G., Barret T., Liu L., et al. RNA cargoes associating with FMRP reveal deficits in cellular functioning in Fmr1 null mice. Neuron. 2003;37:417–431. doi: 10.1016/S0896-6273(03)00034-5. [PubMed] [Cross Ref]
[4] Zalfa F., Giorgi M., Primerano B., Moro A., Di Penta A., Reis S., et al. The fragile X syndrome protein FMRP associates with BC1 RNA and regulates the translation of specific mRNAs at synapses. Cell. 2003;112:317–327. doi: 10.1016/S0092-8674(03)00079-5. [PubMed] [Cross Ref]
[5] Gonzalez-Billault C., Jimenez-Mateos E.M., Caceres A., Diaz-Nido J., Wandosell F., Avila J. Microtubule-associated protein 1B function during normal development, regeneration, and pathological conditions in the nervous system. J Neurobiol. 2004;58:48–59. doi: 10.1002/neu.10283. [PubMed] [Cross Ref]
[6] Zhang Y.Q., Bailey A.M., Matthies H.J., Renden R.B., Smith M.A., Speese S.D., et al. Drosophila fragile X-related gene regulates the MAP1B homolog Futsch to control synaptic structure and function. Cell. 2001;107:591–603. doi: 10.1016/S0092-8674(01)00589-X. [PubMed] [Cross Ref]
[7] Lu R., Wang H., Liang Z., Ku L., O’donnell W.T., Li W., et al. The fragile X protein controls microtubule-associated protein 1B translation and microtubule stability in brain neuron development. Proc Natl Acad Sci USA. 2004;101:15201–15206. doi: 10.1073/pnas.0404995101. [PubMed] [Cross Ref]
[8] Chen L., Yun S.W., Seto J., Liu W., Toth M. The Fragile X mental retardation protein binds and regulates a novel cless of mRNAs containing U rich target sequences. Neuroscience. 2003;120:1005–1017. doi: 10.1016/S0306-4522(03)00406-8. [PubMed] [Cross Ref]
[9] Bakker C.E., Verheij C., Willemsen R., van der Helm R., Oerlemans F., Vermey M., et al. Fmr1 knockout mice: a model to study fragile X mental retardation. Cell. 1994;78:23–33. [PubMed]
[10] Franklin K.B.J., Paxinos G. The Mouse Brain in Stereotaxic Coordinates. 2. San Diego: Academic Press; 2003. pp. 101–129.
[11] Brugg B., Reddy D., Matus A. Attenuation of microtubule-associated protein 1B expression by antisense oligodeoxynucleotides inhibits initiation of neurite outgrowth. Neuroscience. 1993;52:489–496. doi: 10.1016/0306-4522(93)90401-Z. [PubMed] [Cross Ref]
[12] Gonzalez-Billault C., Avila J., Caceres A. Evidence for the role of MAP1B in axon formation. Mol Biol Cell. 2001;12:2087–2098. [PMC free article] [PubMed]
[13] Tucker R.P. The roles of microtubule-associated proteins in brain morphogenesis: a review. Brain Res Rev. 1990;15:101–120. doi: 10.1016/0165-0173(90)90013-E. [PubMed] [Cross Ref]
[14] Montesinos M.L., Foucher I., Conradt M., Mainguy G., Robel L., Prochiantz A., et al. The neuronal microtubule-associated protein 1B is under homeoprotein transcriptional control. J Neurosci. 2001;21:3350–3359. [PubMed]
[15] Ma D., Nothias F., Boyne L.J., Fischer I. Differential regulation of microtubule-associated protein 1B (MAP1B) in rat CNS and PNS during development. J Neurosci Res. 1997;49:319–332. doi: 10.1002/(SICI)1097-4547(19970801)49:3<319::AID-JNR7>3.0.CO;2-F. [PubMed] [Cross Ref]
[16] Darnell J.C., Jensen K.B., Jin P., Brown V., Warren S.T., Darnell R.B. Fragile X mental retardation protein targets G quartet mRNAs important for neuronal function. Cell. 2001;107:489–499. doi: 10.1016/S0092-8674(01)00566-9. [PubMed] [Cross Ref]
[17] Antar L.N., Dictenberg J.B., Plociniak M., Afroz R., Bassell G.J. Localization of FMRP-associated mRNA granules and requirement of microtubules for activity-dependent trafficking in hippocampal neurons. Genes Brain Behav. 2005;4:350–359. doi: 10.1111/j.1601-183X.2005.00128.x. [PubMed] [Cross Ref]
[18] Miyashiro K.Y., Beckel-Mitchener A., Purk T.P., Becker K.G., Barret T., Liu L., et al. RNA cargoes associating with FMRP reveal deficits in cellular functioning in Fmr1 null mice. Neuron. 2003;37:417–431. doi: 10.1016/S0896-6273(03)00034-5. [PubMed] [Cross Ref]
[19] Hinds H.L., Ashley C. T., Sutcliffe J. S., Nelson D.L., Warren S.T., Housman D.E., et al. Tissue specific expression of FMR1 provides evidence for a functional role in fragile X syndrome. Nature Genet. 1993;3:36–43. doi: 10.1038/ng0193-36. [PubMed] [Cross Ref]
[20] Lim J.H., Luo T., Sargent T.D., Fallon J.R. Developmental expression of Xenopus fragile X mental retardation-1 gene. Int J Dev Biol. 2005;49:981–984. doi: 10.1387/ijdb.052070jl. [PubMed] [Cross Ref]
[21] Jin P., Zarnescu D.C., Ceman S., Nakamoto M., Mowrey J., Jongens T.A., et al. Biochemical and genetic interaction between the fragile X mental retardation protein and the microRNA pathway. Nat Neurosci. 2004;7:113–117. doi: 10.1038/nn1174. [PubMed] [Cross Ref]

Articles from Neuroscience Bulletin are provided here courtesy of Springer