Improved β-catenin detection in spinal cord tissue sections: autofluorescence quenching

Authors

  • Dauda Abdullahi (1) Department of Anatomy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia. (2) Department of Anatomy, College of Medical Sciences, Abubakar Tafawa Balewa University Bauchi, Bauchi State, Nigeria.
  • Azlina Ahmad Annuar Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
  • Junedah Sanusi Department of Anatomy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

DOI:

https://doi.org/10.31117/neuroscirn.v3i2.49

Keywords:

spinal cord, β-catenin, autofluorescence quenching, regeneration

Abstract

Experimental studies on spinal cord regeneration are focusing on the windows of opportunity to improve spinal cord microenvironment via spinal-centric repair pathways. One pathway of particular interest is the Wnt/β-catenin signalling pathway which plays a vital role in axonal guidance, synaptic assembly and function, neuronal survival and connectivity after spinal cord trauma to induce repair. Upregulation of β-catenin expression is often taken as evidence of regeneration mechanisms through the Wnt/ β-catenin pathway. However, these studies may not have optimised the staining protocol for β-catenin to enable accurate detection of the protein. Given possible issues with the background or endogenous tissue autofluorescence, there is a need to optimise the protocol further to allow better visualisation of β-catenin. So far, there are no studies which report optimising spinal cord tissues for β-catenin to reduce autofluorescence, and as β-catenin is widely used in spinal cord injury (SCI) and other spinal cord tissue studies, thus it is an important issue to address. To achieve reliable detection and localisation of β-catenin, we utilised sequential quenching techniques using 1% NaBH4 and 1mM CuSO4 in 50mM ammonium acetate buffer to reduce both background and fixative-induced autofluorescence. Our results showed that sequential autofluorescence quenching is crucial in β-catenin detection, and this improved technique indicates that β-catenin is localised in the spinal cord white matter regions. Objective approach for the β-catenin localisation is highly significant as it unravelled an objective identification and illuminate the pattern of distribution of β-catenin for researcher focusing on spinal cord repair studies via the Wnt/β-catenin pathway following SCI.

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Published

2020-05-29

How to Cite

Abdullahi, D., Ahmad Annuar, A. and Sanusi, J. (2020) “Improved β-catenin detection in spinal cord tissue sections: autofluorescence quenching”, Neuroscience Research Notes, 3(2), pp. 4-14. doi: 10.31117/neuroscirn.v3i2.49.

Issue

Section

Technical Notes