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Protein synthetic machinery and mRNA in regenerating tips of spinal cord axons in lamprey.

The Journal of comparative neurology | 2016

Polyribosomes, mRNA, and other elements of translational machinery have been reported in peripheral nerves and in elongating injured axons of sensory neurons in vitro, primarily in growth cones. Evidence for involvement of local protein synthesis in regenerating central nervous system (CNS) axons is less extensive. We monitored regeneration of back-labeled lamprey spinal axons after spinal cord transection and detected mRNA in axon tips by in situ hybridization and microaspiration of their axoplasm. Poly(A)+mRNA was present in the axon tips, and was more abundant in actively regenerating tips than in static or retracting ones. Target-specific polymerase chain reaction (PCR) and in situ hybridization revealed plentiful mRNA for the low molecular neurofilament subunit and β-tubulin, but very little for β-actin, consistent with the morphology of their tips, which lack filopodia and lamellipodia. Electron microscopy showed ribosomes/polyribosomes in the distal parts of axon tips and in association with vesicle-like membranes, primarily in the tip. In one instance, there were structures with the appearance of rough endoplasmic reticulum. Immunohistochemistry showed patches of ribosomal protein S6 positivity in a similar distribution. The results suggest that local protein synthesis might be involved in the mechanism of axon regeneration in the lamprey spinal cord. J. Comp. Neurol. 524:3614-3640, 2016. © 2016 Wiley Periodicals, Inc.

Pubmed ID: 27120118 RIS Download

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This is a list of tools and resources that we have found mentioned in this publication.


BLASTX (tool)

RRID:SCR_001653

Web application to search protein databases using a translated nucleotide query. Translated BLAST services are useful when trying to find homologous proteins to a nucleotide coding region. Blastx compares translational products of the nucleotide query sequence to a protein database. Because blastx translates the query sequence in all six reading frames and provides combined significance statistics for hits to different frames, it is particularly useful when the reading frame of the query sequence is unknown or it contains errors that may lead to frame shifts or other coding errors. Thus blastx is often the first analysis performed with a newly determined nucleotide sequence and is used extensively in analyzing EST sequences. This search is more sensitive than nucleotide blast since the comparison is performed at the protein level.

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S6 Ribosomal Protein (54D2) Mouse mAb (antibody)

RRID:AB_2238583

This monoclonal targets S6 Ribosomal Protein (54D2) Mouse mAb

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Mouse IgG (H&L) Antibody (antibody)

RRID:AB_220347

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Zeiss Axioskop Software (software resource)

RRID:SCR_014587

THIS RESOURCE IS NO LONGER IN SERVCE, documented January 19, 2022. Software used to control the Zeiss Axioskop microscope.

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NIS-Elements (software resource)

RRID:SCR_014329

Microscope imaging software suite used with Nikon products. NIS-Elements includes software applications for advanced and standard research, documentation, confocal microscopy, and high-content analysis.

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RRID:SCR_014333

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Adobe Photoshop (software resource)

RRID:SCR_014199

Software for image processing, analysis, and editing. The software includes features such as touch capabilities, a customizable toolbar, 2D and 3D image merging, and Cloud access and options.

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S6 Ribosomal Protein (54D2) Mouse mAb (antibody)

RRID:AB_2238583

This monoclonal targets S6 Ribosomal Protein (54D2) Mouse mAb

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S6 Ribosomal Protein (54D2) Mouse mAb (antibody)

RRID:AB_2238583

This monoclonal targets S6 Ribosomal Protein (54D2) Mouse mAb

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