Dynein/dynactin is necessary for anterograde transport of Mbp mRNA in oligodendrocytes and for myelination in vivo

Amy L. Herbert, Meng Meng Fu, Catherine M. Drerup, Ryan S. Gray, Breanne L. Harty, Sarah D. Ackerman, Thomas O'Reilly-Pol, Stephen L. Johnson, Alex Nechiporuk, Ben A. Barres, Kelly Monk

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Oligodendrocytes in the central nervous system produce myelin, a lipid-rich, multilamellar sheath that surrounds axons and promotes the rapid propagation of action potentials. A critical component of myelin is myelin basic protein (MBP), expression of which requires anterograde mRNA transport followed by local translation at the developing myelin sheath. Although the anterograde motor kinesin KIF1B is involved in mbp mRNA transport in zebrafish, it is not entirely clear how mbp transport is regulated. From a forward genetic screen for myelination defects in zebrafish, we identified a mutation in actr10, which encodes the Arp11 subunit of dynactin, a critical activator of the retrograde motor dynein. Both the actr10 mutation and pharmacological dynein inhibition in zebrafish result in failure to properly distribute mbp mRNA in oligodendrocytes, indicating a paradoxical role for the retrograde dynein/dynactin complex in anterograde mbp mRNA transport. To address the molecular mechanism underlying this observation, we biochemically isolated reporter-tagged Mbp mRNA granules from primary cultured mammalian oligodendrocytes to show that they indeed associate with the retrograde motor complex. Next, we used live-cell imaging to show that acute pharmacological dynein inhibition quickly arrests Mbp mRNA transport in both directions. Chronic pharmacological dynein inhibition also abrogates Mbp mRNA distribution and dramatically decreases MBP protein levels. Thus, these cell culture and whole animal studies demonstrate a role for the retrograde dynein/dynactin motor complex in anterograde mbp mRNA transport and myelination in vivo.

Original languageEnglish (US)
Pages (from-to)E9153-E9162
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number43
DOIs
StatePublished - Oct 24 2017

Fingerprint

Dyneins
Oligodendroglia
Messenger RNA
Zebrafish
Myelin Sheath
Myelin Basic Protein
Pharmacology
Kinesin
Mutation
Dynactin Complex
Action Potentials
Axons
Central Nervous System
Cell Culture Techniques
Lipids

Keywords

  • Dynactin
  • Dynein
  • MRNA transport
  • Myelination
  • Oligodendrocytes

ASJC Scopus subject areas

  • General

Cite this

Dynein/dynactin is necessary for anterograde transport of Mbp mRNA in oligodendrocytes and for myelination in vivo. / Herbert, Amy L.; Fu, Meng Meng; Drerup, Catherine M.; Gray, Ryan S.; Harty, Breanne L.; Ackerman, Sarah D.; O'Reilly-Pol, Thomas; Johnson, Stephen L.; Nechiporuk, Alex; Barres, Ben A.; Monk, Kelly.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 43, 24.10.2017, p. E9153-E9162.

Research output: Contribution to journalArticle

Herbert, Amy L. ; Fu, Meng Meng ; Drerup, Catherine M. ; Gray, Ryan S. ; Harty, Breanne L. ; Ackerman, Sarah D. ; O'Reilly-Pol, Thomas ; Johnson, Stephen L. ; Nechiporuk, Alex ; Barres, Ben A. ; Monk, Kelly. / Dynein/dynactin is necessary for anterograde transport of Mbp mRNA in oligodendrocytes and for myelination in vivo. In: Proceedings of the National Academy of Sciences of the United States of America. 2017 ; Vol. 114, No. 43. pp. E9153-E9162.
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AU - Herbert, Amy L.

AU - Fu, Meng Meng

AU - Drerup, Catherine M.

AU - Gray, Ryan S.

AU - Harty, Breanne L.

AU - Ackerman, Sarah D.

AU - O'Reilly-Pol, Thomas

AU - Johnson, Stephen L.

AU - Nechiporuk, Alex

AU - Barres, Ben A.

AU - Monk, Kelly

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N2 - Oligodendrocytes in the central nervous system produce myelin, a lipid-rich, multilamellar sheath that surrounds axons and promotes the rapid propagation of action potentials. A critical component of myelin is myelin basic protein (MBP), expression of which requires anterograde mRNA transport followed by local translation at the developing myelin sheath. Although the anterograde motor kinesin KIF1B is involved in mbp mRNA transport in zebrafish, it is not entirely clear how mbp transport is regulated. From a forward genetic screen for myelination defects in zebrafish, we identified a mutation in actr10, which encodes the Arp11 subunit of dynactin, a critical activator of the retrograde motor dynein. Both the actr10 mutation and pharmacological dynein inhibition in zebrafish result in failure to properly distribute mbp mRNA in oligodendrocytes, indicating a paradoxical role for the retrograde dynein/dynactin complex in anterograde mbp mRNA transport. To address the molecular mechanism underlying this observation, we biochemically isolated reporter-tagged Mbp mRNA granules from primary cultured mammalian oligodendrocytes to show that they indeed associate with the retrograde motor complex. Next, we used live-cell imaging to show that acute pharmacological dynein inhibition quickly arrests Mbp mRNA transport in both directions. Chronic pharmacological dynein inhibition also abrogates Mbp mRNA distribution and dramatically decreases MBP protein levels. Thus, these cell culture and whole animal studies demonstrate a role for the retrograde dynein/dynactin motor complex in anterograde mbp mRNA transport and myelination in vivo.

AB - Oligodendrocytes in the central nervous system produce myelin, a lipid-rich, multilamellar sheath that surrounds axons and promotes the rapid propagation of action potentials. A critical component of myelin is myelin basic protein (MBP), expression of which requires anterograde mRNA transport followed by local translation at the developing myelin sheath. Although the anterograde motor kinesin KIF1B is involved in mbp mRNA transport in zebrafish, it is not entirely clear how mbp transport is regulated. From a forward genetic screen for myelination defects in zebrafish, we identified a mutation in actr10, which encodes the Arp11 subunit of dynactin, a critical activator of the retrograde motor dynein. Both the actr10 mutation and pharmacological dynein inhibition in zebrafish result in failure to properly distribute mbp mRNA in oligodendrocytes, indicating a paradoxical role for the retrograde dynein/dynactin complex in anterograde mbp mRNA transport. To address the molecular mechanism underlying this observation, we biochemically isolated reporter-tagged Mbp mRNA granules from primary cultured mammalian oligodendrocytes to show that they indeed associate with the retrograde motor complex. Next, we used live-cell imaging to show that acute pharmacological dynein inhibition quickly arrests Mbp mRNA transport in both directions. Chronic pharmacological dynein inhibition also abrogates Mbp mRNA distribution and dramatically decreases MBP protein levels. Thus, these cell culture and whole animal studies demonstrate a role for the retrograde dynein/dynactin motor complex in anterograde mbp mRNA transport and myelination in vivo.

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