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Editors Selection IGR 13-4

Basic research: In-vivo RGC imaging

Comment by Keith Martin on:

20508 In vivo imaging of murine retinal ganglion cells, Leung CKS; Lindsey JD; Crowston JG et al., Journal of Neuroscience Methods, 2008; 168: 475-478

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Experiments to study the dynamics of retinal ganglion cell (RGC) loss have traditionally required relatively large numbers of animals because RGC quantifications have been performed post mortem and therefore each animal only provided information about one timepoint. In-vivo imaging has been achieved before but has required in-jection of substances into the eye or brain to label RGC, with the inherent risk that the injection or the tracer may affect the process being studied. Leung et al. (120) present a novel in-vivo imaging system where Cyan Fluorescent Protein (CFP) expression under the control of a Thy1 promoter is visualised in the RGC of mice by confocal scanning laser ophthalmoscopy. This method allows in-vivo imaging of RGCs over long periods of time in awake mice with high sensitivity and specificity for RGC compared to the gold standard of retrograde labelling. The resolution achieved is impressive and, although ultrastructural features of the cells are currently difficult to identify, it is to be hoped that this may improve with further refinement of the technique. Although the use of such mice in studies of neuroprotection has obvious attractions, some important issues need to be considered.

A novel method is described that allows that in-vivo imaging of RGCs over long periods of time in awake mice with high sensitivity and specifi city for RGC compared to the gold standard of retrograde labelling

In particular, if expression of Thy1 ceases then RGC may become 'invisible'. This is probably not a significant issue in healthy eyes but becomes important when stress occurs. As previous studies have shown (Schlamp 2001, Huang 2006), Thy1 is down regulated in chronic glaucoma even in RGC with apparently normal morphology. Nevertheless, this is a very promising imaging technique that should facilitate both a greater understanding of the dynamics of RGC loss in conditions such as glaucoma and hopefully a reduction in the number of animals needed to explore such processes.

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