RNA splicing capability of live neuronal dendrites
- J. Glanzer†,‡,
- K. Y. Miyashiro†,‡,
- J.-Y. Sul§,
- L. Barrett§,
- B. Belt†,
- P. Haydon§, and
- J. Eberwine†,¶
- Departments of †Pharmacology and §Neuroscience, University of Pennsylvania School of Medicine, Philadelphia, PA 19104
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Edited by Floyd E. Bloom, The Scripps Research Institute, La Jolla, CA, and approved September 29, 2005 (received for review May 9, 2005)
Abstract
Dendrites are specialized extensions of the neuronal soma that contain components of the cellular machinery involved in RNA and protein metabolism. Several dendritically localized proteins are associated with the precursor-mRNA (pre-mRNA) splicing complex, or spliceosome. Although some spliceosome-related, RNA-binding proteins are known to subserve separate cytoplasmic functions when moving between the nucleus and cytoplasm, little is known about the pre-mRNA splicing capacity of intact dendrites. Here, we demonstrate the presence and functionality of pre-mRNA-splicing components in dendrites. When isolated dendrites are transfected with a chicken δ-crystallin pre-mRNA or luciferase reporter pre-mRNA, splicing junctions clustered at or near expected splice sites are observed. Additionally, in vitro synaptoneurosome experiments show that this subcellular fraction contains a similar complement of splicing factors that is capable of splicing chicken δ-crystallin pre-mRNA. These observations suggest that pre-mRNA-splicing factors found in the dendroplasm retain the potential to promote pre-mRNA splicing.
Footnotes
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↵ ¶ To whom correspondence should be addressed. E-mail: eberwine{at}pharm.med.upenn.edu.
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↵ ‡ J.G. and K.Y.M. contributed equally to this work.
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Author contributions: J.G., K.Y.M., B.B., and J.E. designed research; J.G., K.Y.M., J.-Y.S., L.B., B.B., and J.E. performed research; J.G., K.Y.M., and J.E. contributed new reagents/analytic tools; J.G., K.Y.M., J.-Y.S., L.B., B.B., P.H., and J.E. analyzed data; and J.G., K.Y.M., J.-Y.S., L.B., B.B., P.H., and J.E. wrote the paper.
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Conflict of interest statement: No conflicts declared.
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This paper was submitted directly (Track II) to the PNAS office.
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Abbreviations: pre-mRNA, precursor mRNA; SR, serine-arginine; snRNA, small nuclear RNA; NMD, nonsense-mediated decay; SN, synaptoneurosome; RBP, RNA-binding protein; ISH, in situ hybridization; CDC, chicken δ-crystallin; MAP2, microtubule-associated protein 2; RNP, ribonucleoprotein; PSF, protein-associated splicing factor.
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Freely available online through the PNAS open access option.
- Copyright © 2005, The National Academy of Sciences
