Transport and Localization of Exogenous Myelin Basic Protein MRNA Microinjected into Oligodendrocytes

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1993 Oct 1

PMID 7691830

Citations 167

Authors

K Ainger

D Avossa

F Morgan

S J Hill

C Barry

E Barbarese

J H Carson

Affiliations

Soon will be listed here.

Abstract

We have studied transport and localization of MBP mRNA in oligodendrocytes in culture by microinjecting labeled mRNA into living cells and analyzing the intracellular distribution of the injected RNA by confocal microscopy. Injected mRNA initially appears dispersed in the perikaryon. Within minutes, the RNA forms granules which, in the case of MBP mRNA, are transported down the processes to the periphery of the cell where the distribution again becomes dispersed. In situ hybridization shows that endogenous MBP mRNA in oligodendrocytes also appears as granules in the perikaryon and processes and dispersed in the peripheral membranes. The granules are not released by extraction with non-ionic detergent, indicating that they are associated with the cytoskeletal matrix. Three dimensional visualization indicates that MBP mRNA granules are often aligned in tracks along microtubules traversing the cytoplasm and processes. Several distinct patterns of granule movement are observed. Granules in the processes undergo sustained directional movement with a velocity of approximately 0.2 micron/s. Granules at branch points undergo oscillatory motion with a mean displacement of 0.1 micron/s. Granules in the periphery of the cell circulate randomly with a mean displacement of approximately 1 micron/s. The results are discussed in terms of a multi-step pathway for transport and localization of MBP mRNA in oligodendrocytes. This work represents the first characterization of intracellular movement of mRNA in living cells, and the first description of the role of RNA granules in transport and localization of mRNA in cells.

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References

Colman D, Kreibich G, Frey A, Sabatini D . Synthesis and incorporation of myelin polypeptides into CNS myelin. J Cell Biol. 1982; 95(2 Pt 1):598-608. PMC: 2112951. DOI: 10.1083/jcb.95.2.598. View

Wells B, Cantor C . Ribosome binding by tRNAs with fluorescent labeled 3' termini. Nucleic Acids Res. 1980; 8(14):3229-46. PMC: 324366. DOI: 10.1093/nar/8.14.3229. View

Roach A, Boylan K, Horvath S, Prusiner S, HOOD L . Characterization of cloned cDNA representing rat myelin basic protein: absence of expression in brain of shiverer mutant mice. Cell. 1983; 34(3):799-806. DOI: 10.1016/0092-8674(83)90536-6. View

Rebagliati M, Weeks D, Harvey R, Melton D . Identification and cloning of localized maternal RNAs from Xenopus eggs. Cell. 1985; 42(3):769-77. DOI: 10.1016/0092-8674(85)90273-9. View

Weir M, Kornberg T . Patterns of engrailed and fushi tarazu transcripts reveal novel intermediate stages in Drosophila segmentation. Nature. 1985; 318(6045):433-9. DOI: 10.1038/318433a0. View

Bottenstein J . Growth requirements in vitro of oligodendrocyte cell lines and neonatal rat brain oligodendrocytes. Proc Natl Acad Sci U S A. 1986; 83(6):1955-9. PMC: 323203. DOI: 10.1073/pnas.83.6.1955. View

Kachar B, Behar T, Dubois-Dalcq M . Cell shape and motility of oligodendrocytes cultured without neurons. Cell Tissue Res. 1986; 244(1):27-38. DOI: 10.1007/BF00218378. View

Lawrence J, Singer R . Intracellular localization of messenger RNAs for cytoskeletal proteins. Cell. 1986; 45(3):407-15. DOI: 10.1016/0092-8674(86)90326-0. View

Kristensson K, Holmes K, Duchala C, Zeller N, Lazzarini R, Dubois-Dalcq M . Increased levels of myelin basic protein transcripts in virus-induced demyelination. Nature. 1986; 322(6079):544-7. PMC: 7095299. DOI: 10.1038/322544a0. View

10.

Macdonald P, Ingham P, Struhl G . Isolation, structure, and expression of even-skipped: a second pair-rule gene of Drosophila containing a homeo box. Cell. 1986; 47(5):721-34. DOI: 10.1016/0092-8674(86)90515-5. View

11.

Pfeffer S, Rothman J . Biosynthetic protein transport and sorting by the endoplasmic reticulum and Golgi. Annu Rev Biochem. 1987; 56:829-52. DOI: 10.1146/annurev.bi.56.070187.004145. View

12.

Holmes E, Hermanson G, Cole R, de Vellis J . Developmental expression of glial-specific mRNAs in primary cultures of rat brain visualized by in situ hybridization. J Neurosci Res. 1988; 19(4):389-96, 458-65. DOI: 10.1002/jnr.490190402. View

13.

Fontaine B, Sassoon D, Buckingham M, Changeux J . Detection of the nicotinic acetylcholine receptor alpha-subunit mRNA by in situ hybridization at neuromuscular junctions of 15-day-old chick striated muscles. EMBO J. 1988; 7(3):603-9. PMC: 454363. DOI: 10.1002/j.1460-2075.1988.tb02853.x. View

14.

Berleth T, Burri M, Thoma G, Bopp D, Richstein S, Frigerio G . The role of localization of bicoid RNA in organizing the anterior pattern of the Drosophila embryo. EMBO J. 1988; 7(6):1749-56. PMC: 457163. DOI: 10.1002/j.1460-2075.1988.tb03004.x. View

15.

Gergen J, Butler B . Isolation of the Drosophila segmentation gene runt and analysis of its expression during embryogenesis. Genes Dev. 1988; 2(9):1179-93. DOI: 10.1101/gad.2.9.1179. View

16.

Verity A, Campagnoni A . Regional expression of myelin protein genes in the developing mouse brain: in situ hybridization studies. J Neurosci Res. 1988; 21(2-4):238-48. DOI: 10.1002/jnr.490210216. View

17.

Shiota C, Miura M, Mikoshiba K . Developmental profile and differential localization of mRNAs of myelin proteins (MBP and PLP) in oligodendrocytes in the brain and in culture. Brain Res Dev Brain Res. 1989; 45(1):83-94. DOI: 10.1016/0165-3806(89)90010-2. View

18.

Singer R, Langevin G, Lawrence J . Ultrastructural visualization of cytoskeletal mRNAs and their associated proteins using double-label in situ hybridization. J Cell Biol. 1989; 108(6):2343-53. PMC: 2115600. DOI: 10.1083/jcb.108.6.2343. View

19.

Braun R, Peschon J, Behringer R, Brinster R, Palmiter R . Protamine 3'-untranslated sequences regulate temporal translational control and subcellular localization of growth hormone in spermatids of transgenic mice. Genes Dev. 1989; 3(6):793-802. DOI: 10.1101/gad.3.6.793. View

20.

Okabe S, Hirokawa N . Axonal transport. Curr Opin Cell Biol. 1989; 1(1):91-7. DOI: 10.1016/s0955-0674(89)80043-2. View