Abstract
We used cultured adult mouse retinae as a model system to follow and quantify the retraction of dendrites using diolistic labelling of retinal ganglion cells (RGCs) following explantation. Cell death was monitored in parallel by nuclear staining as ‘labelling’ with RGC and apoptotic markers was inconsistent and exceedingly difficult to quantify reliably. Nuclear staining allowed us to delineate a lengthy time window during which dendrite retraction can be monitored in the absence of RGC death. The addition of brain‐derived neurotrophic factor (BDNF) produced a marked reduction in dendritic degeneration, even when application was delayed for 3 days after retinal explantation. These results suggest that the delayed addition of trophic factors may be functionally beneficial before the loss of cell bodies in the course of conditions such as glaucoma.
Highlights
The adult murine retina provides an attractive model to study detailed structural alterations of neurons following axotomy (Johnson & Martin, 2008; Bull et al, 2011; Guerin et al, 2011; Wood et al, 2011; Denk et al, 2015; White et al, 2015)
The key novel finding here is that delayed application of brainderived neurotrophic factor (BDNF) significantly retards dendritic atrophy of retinal ganglion cell (RGC) following retinal explantation
To ensure that under our experimental conditions, the loss of dendrites can be studied in the absence of RGC death, we first established the time course of cell loss in the ganglion cell layer (GCL)
Summary
The adult murine retina provides an attractive model to study detailed structural alterations of neurons following axotomy (Johnson & Martin, 2008; Bull et al, 2011; Guerin et al, 2011; Wood et al, 2011; Denk et al, 2015; White et al, 2015). While the loss of retinal ganglion cells (RGCs) has typically been used as the primary read-out in this preparation (Bull et al, 2011), fewer studies address the changes that might occur in the dendrites of axotomized RGCs prior to cell loss (see Discussion for a very recent example, Johnson et al, 2016). While Sholl analysis is as such well-established and used extensively to quantify dendritic arbourization in numerous central nervous system (CNS) structures, the procedures allowing unbiased and sparse labelling of individual cells are less straightforward. The RGC morphology varies greatly between cell types in the rodent eye, reflecting the large number of individual cell types
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