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. Author manuscript; available in PMC: 2009 Sep 27.
Published in final edited form as: Epilepsia. 2007 Jul 25;48(11):2023–2046. doi: 10.1111/j.1528-1167.2007.01189.x

Figure 6. Effect of Kcna1 gene deletion on Kv subunit and auxiliary subunit expression.

Figure 6

A) Immunoblot of mouse brain membrane fractions were obtained from two different wild-type (WT1, WT2) and Kcna1-deficient (KO1, KO2) mice (two months old). Proteins (10 μg/lane) were extracted in SDS sample buffer, separated by SDS-PAGE, and analyzed for total Kv1.1 or different related Kv subunits (Kv1.2, Kv1.4, Kvβ2). Numbers at left refer to mobility of molecular weight standards (in kDa). None of the related proteins showed changes in Kv1.1−/− mice compared to controls. Similarly, there were no changes in other Kv (or auxiliary) proteins (e.g., Kv1.3, Kv2.1, Kv3.1 isoform β, Kv4.2, PSD-95, Slo) (not shown).

B) Kv1.4 channel protein immunocytochemistry in hippocampus of a wild-type and a Kv1.1 knock-out (Kv1.1−/−) mouse. In both animals, Kv1.4 is highly expressed in the hilus (H) and s.lucidum (luc) of CA3, reflecting its localization in the mossy fiber axons. This channel subunit is also found in the middle molecular layer (mml) of the dentate gyrus (DG). Like Kv1.1, Kv1.4 is absent from granule cell and CA3 pyramidal cell somata. The ICC profile is consistent with the immunoblot data; there are no obvious changes in Kv1.4 density.

C) Double immunofluorescence staining of wild-type (left panels ) and Kv1.1−/− (right panels) hippocampus. C1, 2. Double immunofluorescence staining for Kv1.1 (red) and Kv1.4 (green). Note loss of Kv1.1 staining in Kv1.1−/− hippocampus, and a lack of any dramatic change in the overall intensity or distribution of Kv1.4 staining.

C3,C4. Double immunofluorescence staining for Kv1.2 (red) and Kv1.4 (green). Note lack of any dramatic change in the overall intensity or distribution of Kv1.2 and Kv1.4 staining in wild-type versus Kv1.1−/− hippocampus (apparent decrease in Kv1.4 expression in the hilus of knock-out mice (lighter green in hilus in C4 compared to C3) is likely due to the increase hilar area in the knock-out, and the consequent “dilution” of Kv1.4-containing mossy fibers).

Scale bars: B: 200 μm; C: 100 μm