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. Author manuscript; available in PMC: 2015 Nov 6.
Published in final edited form as: J Neurol Disord. 2014 Nov 4;2(6):i105. doi: 10.4172/2329-6895.1000i105

Examination of the Blood Brain Barrier integrity in a mouse model of the neurodegenerative Canavan’s disease

Seemin Seher Ahmed 1, Guangping Gao 1,✉
PMCID: PMC4635683  NIHMSID: NIHMS706973  PMID: 26550581

The blood brain barrier (BBB) refers to the complex anatomical barrier in the brain composed of endothelial cells, astroglia, pericytes, perivascular macrophages and basal lamina. Its selectivity controls the entry of substances into the Central Nervous System (CNS) (Bradbury, 1985). BBB disruption affects neurodegeneration (Tomkins et al., 2007) and in some cases can be harnessed to deliver intravenous therapeutics to the CNS. Canavan’s disease (CD) is an autosomal recessive neurodegenerative disease caused by lack of aspartoacylase that causes extensive demyelination. The cause of neurodegeneration has not been conclusively determined. We investigated if BBB integrity is affected in the disease using an aspartoacylase knockout mouse (Matalon et al., 2000). 21 day old wildtype and symptomatic diseased mice (n=3) were injected with Evans blue (EB) (4ml/kg). Control mice were injected with saline or EB combined with mannitol pretreatment (3ml/100g) since mannitol reversibly opens the BBB. 30 minutes later, mice were perfused with PBS, and their brains homogenized. The supernatant was measured for absorbance at 610nm to check the presence of EB using known concentrations as standard. Results showed no difference in the BBB of WT and diseased mice (Figure 1). All mice showed similar levels of EB retention in the blood and liver. Spectrophotometric readings showed no significant difference in BBB permeability between WT and diseased animals (Figure 2). The results indicate that the BBB is not disrupted at least in early pathogenesis of CD. Examination of EB retention in the brain with respect to body weight (Figure 3) however indicated that there may be more to the story since diseased mice weigh almost one-third of their littermates (Ahmed et al., 2013). More animals and several time points along the progression of the disease need to be examined to conclusively establish the effect of the disease on BBB or vice versa.

Figure 1.

Figure 1

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Figure 2.

Figure 2

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Figure 3.

Figure 3

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