Hunting Increases Phosphorylation of Calcium/Calmodulin-Dependent Protein Kinase Type II in Adult Barn Owls

Figure 2

Immunoblotting. Brain homogenates of 7 owls ranging in age from 9 d to 2 yrs were analyzed using antibodies against native (αCaMKII) and phosphorylated (pCaMKII) forms of CaMKII and native CREB1. (a) Top, αCaMKII labeled a single band of expected molecular weight, 50 kD. pCaMKII labeled two distinct bands, phospho-αCaMKII (50 kD) and phospho-βCaMKII (60 kD). GADPH was used as loading control. Protein samples from a single juvenile and single adult in the absence (ctrl) and presence (CaM) of calmodulin confirm the ability of the phosphospecific antibody to detect changes in phosphorylation state. Bottom, normalized intensities for αCaMKII, pCaMKII, and their ratio. For total pCaMKII, black bars correspond to phospho-βCaMKII and open bars to phospho-αCaMKII. (b) Top, CREB1 labeled a single band (in most samples) of expected molecular weight, 43 kD, corresponding to the alpha isoform of CREB1. Hatchlings also expressed lower levels of the delta isoform. Bottom, normalized intensities for αCREB1, total CREB1, and total CREB1 binned for hatchlings (2, 9, and 14 d), fledglings (70 and 75 d), and adults (300 d and 2 yr).

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