Knowledge of the precise timing of myelination is critical to the success of zebrafish-based in vivo screening strategies for potential remyelination therapies. This study provides a systematic review of the timing of myelination in the zebrafish spinal cord and a critique of techniques by which it may be accurately assessed. The onset of myelination was found to be 3 days postfertilization (d.p.f.); earlier than previously reported. This coincided with the dorsal migration and differentiation of oligodendrocytes and the expression of myelin basic protein (Mbp) transcripts and protein. Our data suggests that immunohistochemistry with zebrafish-specific anti-Mbp from 3 d.p.f. is the optimal histological method for myelin visualization, while quantification of myelination is more reliably achieved by quantitative polymerase chain reaction (qPCR) for mbp from 5 d.p.f.. Transgenic fluorescent lines such as olig2:EGFP can be used to assess oligodendrocyte cell number at 3 d.p.f. and the development of new, more specific lines may enable real time visualization of myelin itself. Quantitative ultrastructural analysis revealed that the myelination of zebrafish axons is regulated according to axonal growth and not absolute axonal size. This study confirms the use of the zebrafish larvae as a versatile and efficient in vivo model of myelination and provides a platform on which future myelination screening studies can be based.
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Article Synopsis
- Axon myelination plays a crucial role in refining neuronal circuits by adjusting myelin sheath patterns across different axon types, but the coordination behind this process is still not fully understood.
- Recent studies suggest that neuronal activity and the release of vesicles can stimulate the formation of myelin, and oligodendrocytes express proteins that may aid in their interaction with axons for proper myelination.
- In larval zebrafish, the protein Gephyrin (Gphn) appears to selectively enhance myelin formation on GABAergic axons, with findings showing that in Gphn-deficient larvae, there were longer myelin sheaths on GABAergic axons
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