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  4. Transcriptomes of Injured Lamprey Axon Tips: Single-Cell RNA-Seq Suggests Differential Involvement of MAPK Signaling Pathways in Axon Retraction and Regeneration after Spinal Cord Injury

Transcriptomes of Injured Lamprey Axon Tips: Single-Cell RNA-Seq Suggests Differential Involvement of MAPK Signaling Pathways in Axon Retraction and Regeneration after Spinal Cord Injury

Cells, 2022 · DOI: 10.3390/cells11152320 · Published: July 27, 2022

Spinal Cord InjuryRegenerative MedicineNeurology

Simple Explanation

When an axon is severed, such as from a spinal cord injury, the cell body responds in one of two ways: activating a regenerative response allowing the axon to regrow, or undergoing retrograde death or atrophy. Intra-axonal protein synthesis and degradation play an important role in axon regeneration, allowing the growing axon tip to rapidly respond to its needs despite its long distance from the perikaryon. Lampreys, unlike mammals, can regenerate supraspinal axons after complete spinal cord transection. Researchers micro-aspirated axoplasms from growing, static, and retracting axon tips of spinal cord transected lampreys and performed single-cell RNA-seq to understand the role of local protein synthesis in regenerative vs. degenerative responses.

Study Duration
10-14 days
Participants
260 larval lampreys (Petromyzon marinus), 10–13 cm in length (4–5 years old)
Evidence Level
Not specified

Key Findings

  • 1
    Genes were identified that were upregulated selectively in growing (n = 38), static (20) or retracting tips (18). Among them, map3k2, csnk1e and gtf2h were expressed in growing tips, mapk8(1) was expressed in static tips and prkcq was expressed in retracting tips.
  • 2
    Venn diagrams revealed more than 40 components of MAPK signaling pathways, including jnk and p38 isoforms, which were differentially distributed in growing, static and/or retracting tips.
  • 3
    Real-time q-PCR and immunohistochemistry verified the colocalization of map3k2 and csnk1e in growing axon tips.

Research Summary

This study investigates retrograde signals that trigger regeneration or cell death after axotomy in the CNS, focusing on local protein synthesis in axon tips. Single-cell RNA-seq was performed on micro-aspirated axoplasms from growing, static, and retracting axon tips of spinal cord transected lampreys to identify differentially expressed genes (DEGs). The results support differential roles for different MAPK signals in axon regeneration and retraction, with potential cross-talk between MAPK and circadian rhythm pathways.

Practical Implications

Therapeutic Targets

The selective upregulation of specific MAPK pathways, such as map3k2/map2k4/p38α, may be an effective strategy to improve axon regeneration.

Understanding Axon Dynamics

This research provides a comprehensive view of a complex signaling network involving different subsets and isoforms in axon elongation and retraction, advancing the understanding of post-injury axon dynamics.

Regenerative Failure and Cell Death

Genes closely associated with axon retraction (e.g., dcaf17, prkcq, cdk7, rxfp3, oprm1, grm7, and sdc2) may also be involved in signaling regenerative failure and delayed cell death.

Study Limitations

  • 1
    The descriptive results obtained in this study require validation by molecular manipulations.
  • 2
    Lamprey, with its very long life cycle, may not be the best animal in which to test the functions of discovered local mRNAs.
  • 3
    It would be very difficult to obtain the information contained in the present report in species such as mice.

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