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Updated: May 28, 2026
Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR
Published on: February 6, 2026
Limited availability of ZBP1 restricts axonal mRNA localization and nerve regeneration capacity
Christopher J Donnelly1, Dianna E Willis, Mei Xu
1Department of Biological Sciences, University of Delaware, Newark, USA.
Abstract:
Subcellular localization of mRNAs is regulated by RNA-protein interactions. Here, we show that introduction of a reporter mRNA with the 3'UTR of β-actin mRNA competes with endogenous mRNAs for binding to ZBP1 in adult sensory neurons. ZBP1 is needed for axonal localization of β-actin mRNA, and introducing GFP with the 3'UTR of β-actin mRNA depletes axons of endogenous β-actin and GAP-43 mRNAs and attenuates both in vitro and in vivo regrowth of severed axons. Consistent with limited levels of ZBP1 protein in adult neurons, mice heterozygous for the ZBP1 gene are haploinsufficient for axonal transport of β-actin and GAP-43 mRNAs and for regeneration of peripheral nerve. Exogenous ZBP1 can rescue the RNA transport deficits, but the axonal growth deficit is only rescued if the transported mRNAs are locally translated. These data support a direct role for ZBP1 in transport and translation of mRNA cargos in axonal regeneration in vitro and in vivo.
Insights
Z-binding protein 1 (ZBP1) is crucial for transporting mRNAs in neurons, enabling axonal regeneration. Limited ZBP1 impairs nerve repair, but restoring ZBP1 and local mRNA translation can promote regrowth.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Subcellular mRNA localization is vital for neuronal function, controlled by RNA-protein interactions.
- Axonal transport of specific mRNAs, like β-actin mRNA, is essential for neuronal growth and repair.
Purpose of the Study:
- To investigate the role of Z-binding protein 1 (ZBP1) in the axonal localization of mRNAs and its impact on axonal regeneration.
- To determine if ZBP1 levels and local mRNA translation are critical for peripheral nerve regeneration.
Main Methods:
- Utilized reporter mRNAs and endogenous mRNAs in adult sensory neurons to study ZBP1 binding competition.
- Assessed the effects of ZBP1 manipulation on mRNA levels in axons and in vitro/in vivo axonal regrowth.
- Examined ZBP1 haploinsufficiency in mice and the rescue potential of exogenous ZBP1 and local mRNA translation.
Main Results:
- Reporter mRNA with β-actin 3'UTR competes with endogenous mRNAs for ZBP1 binding.
- Reduced ZBP1 levels deplete axons of key mRNAs (β-actin, GAP-43) and impair axonal regrowth.
- ZBP1 haploinsufficiency in mice leads to deficits in axonal mRNA transport and peripheral nerve regeneration.
- Exogenous ZBP1 rescues RNA transport, but axonal growth recovery requires local mRNA translation.
Conclusions:
- ZBP1 plays a direct role in the axonal transport of mRNAs essential for regeneration.
- Local translation of transported mRNAs is critical for successful axonal growth following nerve injury.
- Findings highlight ZBP1 as a key regulator of mRNA dynamics in axonal regeneration.
