मस्तिष्क संबंधी विकार

Zebrafish Whole Mount Staining Using Luxol Fast Blue Stain

Abstract

Waseem Hasan 1 , Mazen Shafea 1 , Doua Abdulrahman 1 and Sahar Daas 1,2*

Background: Neurological disorders prevalence burdens the healthcare system. These neurological disorders are defined as functional impairments that affect the nervous system; the clinical neuropathological manifestations can present structural, biochemical, or electrical abnormalities.

Zebrafish disease models have been used extensively to study human neuropathological and behavioral disorders due to the unique in vivo live real-time visualization of the developing nervous system within the transparent model.

Myelin is a lipoprotein of a fatty multilayered membrane that surrounds the axons of the nerve cells. The myelination of the axons increases the electrical impulse speed rate through the nervous system and indicates the proper function of the nervous system.

In zebrafish model, histological procedures are employed to study the nervous system. However, the proper orientation and serial sections should be obtained the neuroanatomy of the brain at the cellular level. Consequently, it is challenging to examine the structural alterations in real dimensional developing brain.
Luxol fast blue is used to stain myelin sheath which is rich in phospholipids. This stain is used to differentiate white matter (rich in myelin) from the grey matter in brain tissue sections.

Results: Here, we developed a new method of whole-mount staining that optimize the penetration of staining regents of luxol fast blue into the white and gray matter of zebrafish larvae developing brain with decrease non-specific binding and background. Our result showed the clear anatomical structures of the central nervous system as a utility for experimental neuropathology in zebrafish model.

Conclusion: Our developed method provides three-dimensional visualization of the developing central nervous system of zebrafish; this is useful to characterize neuron demyelination in corresponding disease model.