Abstract
Targeting of mRNAs to distinct subcellular regions occurs in all polarized cells. The mechanisms by which RNA transport occurs are poorly understood. With the advent of RNA amplification methodologies and expression profiling it is now possible to catalogue the RNAs that are targeted to particular subcellular regions. In particular, neurons are polarized cells in which dendrites receive signals from presynaptic neurons. Upon stimulation (information receipt) the dendrite processes the information such that an immediate dendritic response is generated as well as a longer-term somatic response. The integrated cellular response results in a signal that can be propagated through the axon to the next post-synaptic neuron. Much previous work has shown that mRNAs can be localized in dendrites and that local translation in dendrites can occur. In this chapter the methods for analysis of RNAs that are localized to dendrites are reviewed and a partial list of dendritically localized RNAs is presented. This information may be useful in identifying RNA regulatory regions that are responsible for specifying rate of RNA transport and the dendritic sites at which targeted RNAs dock so that they can be translated.
Similar content being viewed by others
REFERENCES
Tiedge, H. and Brosius, J. 1996. Translational machinery in dendrites of hippocampal neurons in culture. J. Neurosci. 16(22):7171-7181.
Steward, O. and Levy, W. B. 1982. Preferential localization of polyribosomes under the base of dendritic spines in granule cells of the dentale gyrus. J. Neurosci. 2(3):284-291.
Kiss, J. 1977. Synthesis and transport of newly formed proteins in dendrites of rat hippocampal pyramid cells. An electron microscope autoradiographic study. Brain Res. 124(2):237-250.
Weiler, I. J., Wang X., and Greenough, W. T. 1994. Synapse-activated protein synthesis as a possible mechanism of plastic neural change. Prog. Brain 100:189-194.
Crino, P. B. and Eberwine, J. 1996. Molecular characterization of the dendritic growth cone: regulated mRNA transport and local protein synthesis. Neuron 17(6):1173-1187.
Eberwine, J., Miyashiro, K., Kacharmina, J. E., and Job, C. 2001. Local translation of classes of mRNAs that are targeted to neuronal dendrites. Proc. Natl. Acad. Sci. USA 98(13):7080-7085.
Job, C. and Eberwine, J. 2001. Localization and translation of mRNA in dendrites and axons. Nat. Rev. Neurosci. 2(12):889-898.
Job, C. and Eberwine, J. 2001. Identification of sites for exponential translation in living dendrites. Proc. Natl. Acad. Sci. USA 98(23):13037-13042.
Kang, H. and Schuman, E. M. 1996. A requirement for local protein synthesis in neurotrophin-induced hippocampal synaptic plasticity. Science 273(5280):1402-1406.
Huber, K. M., Kayser, M. S., and Bear, M. F. 2000. Role for rapid dendritic protein synthesis in hippocampal mGluR-dependent long-term depression. Science 288(5469):1254-1257.
Spencer, C. M. and Eberwine, J. 1999. Cytoplasmic proteins interact with a translational control element in the protein-coding region of proopiomelanocortin mRNA. DNA Cell Biol. 18(1):39-49.
Eberwine, J. H., Spencer, C., Newell, D., and Hoffman, A. R. 1993. mRNA structure, in situ, as assessed by microscopic techniques. Microsc. Res. Tech. 25(1):19-28.
Roberts, J. L., Eberwine, J. H., and Gee, C. E. 1983. Analysis of POMC gene expression by transcription assay and in situ hybridization histochemistry. Cold Spring Harb. Symp. Quant. Biol. 48 Pt. 1:385-391.
Tecott, L. H., Barchas, J. D., and Eberwine, J. H. 1988. In situ transcription: specific synthesis of complementary DNA in fixed tissue sections. Science 240(4859):1661-1664.
Saiki, R. K., Bugawan, T. L., Horn, G. T., Mullis, K. B., and Erlich, H. A. 1986. Analysis of enzymatically amplified betaglobin and HLA-DQ alpha DNA with allele-specific oligonucleotide probes. Nature 324(6093):163-166.
Hagen-Mann, K. and Mann, W. 1995. RT-PCR and alternative methods to PCR for in vitro amplification of nucleic acids. Exp. Clin. Endocrinol. Diabetes 103(3):150-155.
Eberwine, J., Yeh, H., Miyashiro, K., Cao, Y., Nair, S., Finnell, R., Zettel, M., and Coleman, P. 1992. Analysis of gene expression in single live neurons. Proc. Natl. Acad. Sci. USA 89(7):3010-3014.
Van Gelder, R. N., von Zastrow, M. E., Yool, A., Dement, W. C., Barchas, J. D., and Eberwine, J. H. 1990. Amplified RNA synthesized from limited quantities of heterogeneous cDNA. Proc. Natl. Acad. Sci. USA 87(5):1663-1667.
Luo, L., Salunga, R. C., Guo, H., Bittner, A., Joy, K. C., Galindo, J. E., Xiao, H., Rogers, K. E., Wan, J. S., Jackson, M. R., and Erlander, M. G. 1999. Gene expression profiles of laser-captured adjacent neuronal subtypes. Nat. Med. 5(1):117-122.
Zarrinkar, P. P., Mainquist, J. K., Zamora, M., Stern, D., Welsh, J. B., Sapinoso, L. M., Hampton, G. M., and Lockhart, D. J. 2001. Arrays of arrays for high-throughput gene expression profiling. Genome Res. 11(7):1256-1261.
Miyashiro, K., Dichter, M., and Eberwine, J. 1994. On the nature and differential distribution of mRNAs in hippocampal neurites: implications for neuronal functioning. Proc. Natl. Acad. Sci. USA 91(23):10800-10804.
Steward, O., Wallace, C. S., Lyford, G. L., and Worley, P. F. 1998. Synaptic activation causes the mRNA for the IEG Arc to localize selectively near activated postsynaptic sites on dendrites. Neuron 21(4):741-751.
Steward, O. and Falk, P. M. 1985. Polyribosomes under developing spine synapses: growth specializations of dendrites at sites of synaptogenesis. J. Neurosci. Res. 13(1-2):75-88.
Torre, E. R. and Steward, O. 1996. Protein synthesis within dendrites: glycosylation of newly synthesized proteins in dendrites of hippocampal neurons in culture. J. Neurosci. 16(19):5967-5978.
Kacharmina, J. E., Job, C., Crino, P., and Eberwine, J. 2000. Stimulation of glutamate receptor protein synthesis and membrane insertion within isolated neuronal dendrites. Proc. Natl. Acad. Sci. USA 97(21):11545-11550.
Meshorer, E., et al. 2002. Alternative splicing and neuritic mRNA translocation under long-term neuronal hypersensitivity. Science 295(5554):508-512.
Kleiman, R., Banker, G., and Steward, O. 1994. Development of subcellular mRNA compartmentation in hippocampal neurons in culture. J. Neurosci. 14(3 Pt 1):1130-1140
Laurent-Demir, C., et al. 2000. Differential regulation of Ca(2+)-calmodulin stimulated and Ca(2+)-insensitive adenylyl cyclase messenger RNA in intact and denervated mouse hippocampus. Neuroscience 96(2):267-274.
Strong, M. J., et al. 1990. The temporal expression of amyloid precursor protein mRNA in vitro in dissociated hippocampal neuron cultures. Exp. Neurol. 109(2):171-179.
Lyford, G. L., et al. 1995. Arc, a growth factor and activity-regulated gene, encodes a novel cytoskeleton-associated protein that is enriched in neuronal dendrites. Neuron. 14(2):433-445.
Tongiorgi, E., Righi, M., and Cattaneo, A. 1997. Activity-dependent dendritic targeting of BDNF and TrkB mRNAs in hippocampal neurons. J. Neurosci. 17(24):9492-9505.
Berry, F. B., and Brown, I. R. 1996. CaMI mRNA is localized to apical dendrites during postnatal development of neurons in the rat brain. J. Neurosci. Res. 43(5):565-575.
Burgin, K. E., et al. 1990. In situ hybridization histochemistry of Ca2+/calmodulin-dependent protein kinase in developing rat brain. J. Neurosci. 10(6):1788-1798.
Eastwood, S. L. and Harrison, P. J. 2000. Complexin I and II mRNAs in the human hippocampus. Mol. Psychiatry 5(4):341.
Hevner, R. F., and Wong-Riley, M. T. 1991. Neuronal expression of nuclear and mitochondrial genes for cytochrome oxidase (CO) subunits analyzed by in situ hybridization: comparison with CO activity and protein. J. Neurosci. 11(7):1942-1958.
Neuner-Jehle, M., et al. 1996. Characterization and sleep deprivation-induced expression modulation of dendrin, a novel dendritic protein in rat brain neurons. J. Neurosci. Res. 46(2):138-151.
Herb, A., et al. 1997. Prominent dendritic localization in forebrain neurons of a novel mRNA and its product, dendrin. Mol. Cell Neurosci. 8(5):367-374.
Ishimoto, T., et al. 2000. Dendritic translocation of the rat ferritin H chain mRNA. Biochem. Biophys. Res. Commun. 272(3):789-793.
Sperk, G., et al. 1998. Expression of GABA(A) receptor subunits in the hippocampus of the rat after kainic acid-induced seizures. Epilepsy Res. 32(1-2):129-139.
Chicurel, M. E., Terrian, D. M., and Potter, H. 1993. mRNA at the synapse: analysis of a synaptosomal preparation enriched in hippocampal dendritic spines. J. Neurosci. 13(9):4054-4063.
Landry, C. F., et al. 1994. Cellular influences on RNA sorting in neurons and glia: an in situ hybridization histochemical study. Brain Res. Mol. Brain Res. 27(1):1-11.
Racca, C., Gardiol, A., and Triller, A. 1997. Dendritic and postsynaptic localizations of glycine receptor alpha subunit mRNAs. J. Neurosci. 17(5):1691-1700.
Watson, J. B., et al. 1994. G-protein gamma 7 subunit is selectively expressed in medium-sized neurons and dendrites of the rat neostriatum. J. Neurosci. Res. 39(1):108-116.
Link, W., et al. 1995. Somatodendritic expression of an immediate early gene is regulated by synaptic activity. Proc. Natl. Acad. Sci. USA 92(12):5734-5738.
Foster, J. A., and Brown, I. R. 1996. Intracellular localization of heat shock mRNAs (hsc70 and hsp70) to neural cell bodies and processes in the control and hyperthermic rabbit brain. J. Neurosci. Res. 46(6):652-665.
Furuichi, T., et al. 1993. Widespread expression of inositol 1,4,5-triphosphate receptor type 1 gene (Insp3r1) in the mouse central nervous system. Receptors Channels 1(1):11-24.
Bloch, B., et al. 1990. Presence of neuropeptide messenger RNAs in neuronal processes. Neurosci. Lett. 109(3):259-264.
Severt, W. L., et al. 1999. The suppression of testis-brain RNA binding protein and kinesin heavy chain disrupts mRNA sorting in dendrites. J. Cell. Sci. 112(Pt 21):3691-3702.
Garner, C. C., Tucker, R. P., and Matus, A. 1988. Selective localization of messenger RNA for cytoskeletal protein MAP2 in dendrites. Nature 336(6200):674-677.
Grant, A. L., et al. 1996. Characterization of the rat hippocalcin gene: the 5´ flanking region directs expression to the hippocampus. Neuroscience 75(4):1099-1115.
Matsuoka, Y., et al. 2002. A chromodomain-containing nuclear protein, MRG15 is expressed as a novel type of dendritic mRNA in neurons. Neurosci. Res. 42(4):299-308.
Paradies, M. A., and Steward, O. 1997. Multiple subcellular mRNA distribution patterns in neurons: a nonisotopic in situ hybridization analysis. J. Neurobiol. 33(4):473-493.
Krishna, N. S., et al. 1995. Human olfactory receptor neurons contain OMP mRNA in their dendritic and axonal processes. Neuroreport 6(5):817-821.
Bian, F., et al. 1996. Differential mRNA transport and the regulation of protein synthesis: selective sensitivity of Purkinje cell dendritic mRNAs to translational inhibition. Mol. Cell Neurosci. 7(2):116-133.
Moriya, M., and Tanaka, S. 1994. Prominent expression of protein kinase C (gamma) mRNA in the dendrite-rich neuropil of mice cerebellum at the critical period for synaptogenesis. Neuroreport. 5(8):929-932.
Gelot, A., et al. 1996. Alpha-brain spectrin mRNA belongs to the population of intradendritically transported mRNAs. Neuroreport. 8(1):113-116.
Kosik, K. S., et al. 1989. Tau in situ hybridization in normal and Alzheimer brain: localization in the somatodendritic compartment. Ann. Neurol. 26(3):352-361.
Dumas, S., et al. 1990. Tyrosine hydroxylase gene expression in human ventral mesencephalon: detection of tyrosine hydroxylase messenger RNA in neurites. J. Neurosci. Res. 25(4):569-575.
Mohr, E., Morris, J. F., and Richter, D. 1995. Differential subcellular mRNA targeting: deletion of a single nucleotide prevents the transport to axons but not to dendrites of rat hypothalamic magnocellular neurons. Proc. Natl. Acad. Sci. USA 92(10):4377-4381.
Eberwine, J., et al. 2001. Transcription factors in dendrites: dendritic imprinting of the cellular nucleus. Results Probl. Cell Differ. 34:57-68.
Miyashiro, K. and Eberwine, J. unpublished observations.
Bodian, D. 1965. A suggestive relationship of nerve cell RNA with specific synaptic sites. Proc. Natl. Acad. Sci. USA 53:418-425.
Miyashiro, K., Finnel, R. H., and Eberwine, J. Map kinase pathways regulate ank mRNA localization to neuronal dendrites. Submitted.
Therianos, S., et al. Transcription factor mRNA and protein in dendrites: Interneuronal transport of Nkx-2.2. Submitted.
Miyashiro, K., et al. RNA cargoes associating with FMRP reveal deficits in cellular frunctionin in FMR1 null mice. Submitted.
Crino, P., et al. 1998. Presence and phosphorylation of transcription factors in dendrites. Proc. Natl. Acad. Sci. USA 95:2313-2318.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Eberwine, J., Belt, B., Kacharmina, J.E. et al. Analysis of Subcellularly Localized mRNAs Using in Situ Hybridization, mRNA Amplification, and Expression Profiling. Neurochem Res 27, 1065–1077 (2002). https://doi.org/10.1023/A:1020956805307
Issue Date:
DOI: https://doi.org/10.1023/A:1020956805307