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Echinoderm microtubule-associated protein -like protein 5 in anterior temporal neocortex of patients with intractable epilepsy.
Sun JJ
,
Huang M
,
Xiao F
,
Xi ZQ
.
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OBJECTIVES: EMAP-like Protein 5 (EML5) is a new echinoderm microtubule-associated protein that is expressed in the central nervous system. The aim of this study was to investigate the expression profile of EML5 in the anterior temporal neocortex of patients presenting with intractable epilepsy (IE).
MATERIALS AND METHODS: Western blot assays were performed to determine EML5 expression in 36 surgically resected anterior temporal neocortices of patients with IE and eight control tissues. Immunohistochemistry and immunofluorescence were employed to explore protein expression in IE.
RESULTS: EML5 was highly expressed in both neurons and glial cells of the anterior temporal neocortex of IE patients, whereas only low levels of EML5 were detected in control brain tissues. Western blotting showed an enhanced expression of EML5 protein in the anterior temporal neocortex of IE (optical density (OD) = 1.8030 ± 0.1335/1.1852 ± 0.2253, P<0.05) compared with normal control tissues.
CONCLUSION: The results demonstrate that highly expressed EML5 in the neurons and glial cells of the cortex of patients with epilepsy is associated with microtubular dysfunction after frequent and recurrent seizures.
Figure 1. FQ-PCR analysis of EML5 mRNA in the anterior temporal lobe of IE and control patients. (A) SYBR fluorescence amplification curve; (B) The relative expression of EML5 mRNA in IE patients was higher than that in the control (greater than a five-fold change)
Figure 2. Immunohistochemical staining of the anterior temporal neocortex with antibody targeted to EML5. EML5 was abnormally expressed in the temporal neocortex of IE patients (A, C, and D), whereas minimal immunoreactive protein was found in the normal temporal neocortex (B, and E); a, neuron; b, glial cell; DAB staining (A, and B); Immunofluorescence staining (C, D, and E) (Bar: A, B, C, and E=75 μm; D=37.5 μm)
Figure 3. A: Western blot analysis of the EML5 expression in the anterior temporal lobe of patients with IE. A representative EML5 immunoblot that was loaded with homogenates from the control and IE cases is shown. Molecular weight markers (MW, kDa) are indicated on the left. IE, intractable epilepsy; Ctr, control. B: Semi-quantitative analysis of EML5 in brain homogenates of IE (P<0.05)
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