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Isolation of mouse neuritic mRNAs

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Abstract

Impaired local protein translation at postsynaptic sites has been hypothesized to be the cause of several neurological disorders such as fragile X syndrome, neurofibromatosis-1, Rett syndrome, and other syndromic and non-specific forms of mental retardation. Identification of which mRNAs are present in dendrites and the identification of the molecular pathways that they promote will be imperative to the understanding of the neuropathology of these diseases. Since mouse models are the most widely used animal models of human diseases we developed a cell culture based technique to isolate mRNAs from mouse neurites.

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References

  • Burgin KE, Waxham MN, Rickling S, Westgate SA, Mobley WC, Kelly PT (1990) In situ hybridization histochemistry of Ca2+/calmodulin-dependent protein kinase in developing rat brain. J Neurosci 10:1788–1798

    PubMed  CAS  Google Scholar 

  • Chamak B, Prochiantz A (1989) Influence of extracellular matrix proteins on the expression of neuronal polarity. Development 106:483–491

    PubMed  CAS  Google Scholar 

  • Che S, Ginsberg SD (2004) Amplification of RNA transcripts using terminal continuation. Lab Invest 84:131–137

    Article  PubMed  CAS  Google Scholar 

  • Comery TA, Harris JB, Willems PJ, Oostra BA, Irwin SA, Weiler IJ, Greenough WT (1997) Abnormal dendritic spines in fragile X knockout mice: Maturation and pruning deficits. Proc Natl Acad Sci USA 94:5401–5404

    Article  PubMed  CAS  ADS  Google Scholar 

  • Craven RA, Banks RE (2002). Use of laser capture microdissection to selectively obtain distinct populations of cells for proteomic analysis. Methods Enzymol 356:33–49

    PubMed  CAS  Google Scholar 

  • Demetrick DJ, Murthy SK, DiFrancesco LM (2002) Fluorescence in situ hybridization of LCM-isolated nuclei from paraffin sections. Methods Enzymol 356:63–69

    PubMed  CAS  Google Scholar 

  • Garner CC, Tucker RP, Matus A (1988). Selective localization of messenger RNA for cytoskeletal protein MAP2 in dendrites. Nature 336:674–677

    Article  PubMed  CAS  ADS  Google Scholar 

  • Gazzaley AH, Benson DL, Huntley GW, Morrison JH (1997) Differential subcellular regulation of NMDAR1 protein and mRNA in dendrites of dentate gyrus granule cells after perforant path transection. J Neurosci 17:2006–2017

    PubMed  CAS  Google Scholar 

  • Hahn S, Zhong XY, Holzgreve W (2002). Single cell PCR in laser capture microscopy. Methods Enzymol 356:295–301

    PubMed  CAS  Google Scholar 

  • Huang LE, Luzzi V, Ehrig T, Holtschlag V, Watson MA (2002) Optimized tissue processing and staining for laser capture microdissection and nucleic acid retrieval. Methods Enzymol 356:49–62

    Article  PubMed  CAS  Google Scholar 

  • Huber KM, Gallagher SM, Warren ST, Bear MF (2002) Altered synaptic plasticity in a mouse model of fragile X mental retardation. Proc Natl Acad Sci USA 99:7746–7750

    Article  PubMed  CAS  ADS  Google Scholar 

  • Irwin SA, Idupulapati M, Gilbert ME, Harris JB, Chakravarti AB, Rogers EJ, Crisostomo RA, Larsen BP, Mehta A, Alcantara CJ, et al. (2002) Dendritic spine and dendritic field characteristics of layer V pyramidal neurons in the visual cortex of fragile-X knockout mice. Am J Med Genet 111:140–146

    Article  PubMed  Google Scholar 

  • Irwin SA, Patel B, Idupulapati M, Harris JB, Crisostomo RA, Larsen BP, Kooy F, Willems PJ, Cras P, Kozlowski PB, et al. (2001) Abnormal dendritic spine characteristics in the temporal and visual cortices of patients with fragile-X syndrome: a quantitative examination. Am J Med Genet 98:161–167

    Article  PubMed  CAS  Google Scholar 

  • Kang H, Schuman EM (1996) A requirement for local protein synthesis in neurotrophin-induced hippocampal synaptic plasticity. Science 273:1402–1406

    PubMed  CAS  ADS  Google Scholar 

  • Kleiman R, Banker G, Steward O (1994) Development of subcellular mRNA compartmentation in hippocampal neurons in culture. J Neurosci 14:1130–1140

    PubMed  CAS  Google Scholar 

  • Kleinman HK, McGarvey ML, Liotta LA, Robey PG, Tryggvason K, Martin GR (1982) Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry 21:6188–6193

    Article  PubMed  CAS  Google Scholar 

  • Koekkoek SKE, Yamaguchi K, Milojkovic BA, Dortland BR, Ruigrok TJH, Maex R, Vellema M, Smit AE, van der Werf F, Bakker CE, et al. (2005) Enhanced LTD at enlarged Purkinje cell spines causes motor learning deficits in fragile X syndrome. Neuron (in press)

  • Kwon H, Menon V, Eliez S, Warsofsky IS, White CD, Dyer-Friedman J, Taylor AK, Glover GH, Reiss AL (2001) Functional neuroanatomy of visuospatial working memory in fragile x syndrome: relation to behavioral and molecular measures. Am J Psychiatry 158:1040–1051

    Article  PubMed  CAS  Google Scholar 

  • Laggerbauer B, Ostareck D, Keidel EM, Ostareck-Lederer A, Fischer U (2001) Evidence that fragile X mental retardation protein is a negative regulator of translation. Hum Mol Genet 10:329–338

    Article  PubMed  CAS  Google Scholar 

  • Lemke G (2001) Glial control of neuronal development. Annu Rev Neurosci 24:87–105

    Article  PubMed  CAS  Google Scholar 

  • Li J, Pelletier MR, Perez Velazquez JL, Carlen PL (2002) Reduced Cortical Synaptic Plasticity and GluR1 Expression Associated with Fragile X Mental Retardation Protein Deficiency. Mol Cell Neurosci 19:138–151

    Article  PubMed  CAS  Google Scholar 

  • Li YL, Sato M, Kojima N, Miura M, Senoo H (1999) Regulatory role of extracellular matrix components in expression of matrix metalloproteinases in cultured hepatic stellate cells. Cell Struct Funct 24:255–261

    Article  PubMed  CAS  Google Scholar 

  • Lu R, Wang H, Liang Z, Ku L, O’Donnell WT, Li W, Warren ST, Feng Y (2004) The fragile X protein controls microtubule-associated protein 1B translation and microtubule stability in brain neuron development. Proc Natl Acad Sci USA 101:15201–15206

    Article  PubMed  CAS  ADS  Google Scholar 

  • Lyford GL, Yamagata K, Kaufmann WE, Barnes CA, Sanders LK, Copeland NG, Gilbert DJ, Jenkins NA, Lanahan AA, Worley PF (1995) Arc, a growth factor and activity- regulated gene, encodes a novel cytoskeleton-associated protein that is enriched in neuronal dendrites. Neuron 14: 433–445

    Article  PubMed  CAS  Google Scholar 

  • Martin KC (2004) Local protein synthesis during axon guidance and synaptic plasticity. Curr Opin Neurobiol 14:305–310

    Article  PubMed  CAS  ADS  Google Scholar 

  • Martin KC, Casadio A, Zhu H, Yaping E, Rose JC, Chen M, Bailey CH, Kandel ER (1997) Synapse-specific, long-term facilitation of aplysia sensory to motor synapses: a function for local protein synthesis in memory storage. Cell 91:927–938

    Article  PubMed  CAS  Google Scholar 

  • Mayer A, Stich M, Brocksch D, Schutze K, Lahr G (2002) Going in vivo with laser microdissection. Methods Enzymol 356:25–33

    PubMed  Google Scholar 

  • Miller S, Yasuda M, Coats JK, Jones Y, Martone ME, Mayford M (2002) Disruption of dendritic translation of CaMKIIalpha impairs stabilization of synaptic plasticity and memory consolidation. Neuron 36:507–519

    Article  PubMed  CAS  Google Scholar 

  • Ohyama H, Mahadevappa M, Luukkaa H, Todd R, Warrington JA, Wong DT (2002). Use of laser capture microdissection-generated targets for hybridization of high-density oligonucleotide arrays. Methods Enzymol 356:323–333

    PubMed  CAS  Google Scholar 

  • Persson A, Backvall H, Ponten F, Uhlen M, Lundeberg J (2002). Single cell gene mutation analysis using laser-assisted microdissection of tissue sections. Methods Enzymol 356:334–343

    PubMed  CAS  Google Scholar 

  • Sherff CM, Carew TJ (1999) Coincident induction of long-term facilitation in Aplysia: cooperativity between cell bodies and remote synapses. Science 285:1911–1914

    Article  PubMed  CAS  Google Scholar 

  • Steward O, Levy WB (1982) Preferential localization of polyribosomes under the base of dendritic spines in granule cells of the dentate gyrus. J Neurosci 2:284–291

    PubMed  CAS  Google Scholar 

  • Steward O, Worley P (2002) Local synthesis of proteins at synaptic sites on dendrites: role in synaptic plasticity and memory consolidation? Neurobiol Learn Mem 78:508–527

    Article  PubMed  CAS  Google Scholar 

  • Torre ER, Steward O (1992). Demonstration of local protein synthesis within dendrites using a new cell culture system that permits the isolation of living axons and dendrites from their cell bodies. J Neurosci 12:762–772

    PubMed  CAS  Google Scholar 

  • Wittliff JL, Erlander MG (2002) Laser capture microdissection and its applications in genomics and proteomics. Methods Enzymol 356:12–25

    PubMed  CAS  Google Scholar 

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Correspondence to Rob Willemsen.

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Reis, S.A., Oostra, B.A. & Willemsen, R. Isolation of mouse neuritic mRNAs. J Mol Hist 37, 79–86 (2006). https://doi.org/10.1007/s10735-006-9036-7

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  • DOI: https://doi.org/10.1007/s10735-006-9036-7

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