Figure 1.

P modulation of CaMKII basal and total activities in the presence and absence of CaMKII inhibitor KN-93 (A) and Western immunoblotting for CaMKII protein (B) in the VMN. Ovariectomized rats with chronic in-dwelling stainless steel cannulae in the third cerebral ventricle were primed with EB or vehicle (V) sc. Forty-eight hours later, P was administered icv and killed 30 min later. A, The VMN tissue homogenates were assayed for CaMKII basal and total activities in the absence and presence of CaMKII inhibitor KN-93 (inhibitor) infused (icv) 30 min before P or V treatment. The CaMKII basal and total activities are expressed as percentage of vehicle control (mean ± sem; 100%). The CaMKII activity was normalized to the vehicle in the presence of KN-93 and expressed as percentage of the respective control (mean ± sem; 100%). The specific activity of V control in the presence of KN-93 was 0.2 pmol/min·mg and in the absence of KN-93 was 12.4 pmol/min·mg. Statistical analysis using ANOVA followed by Dunnett’s or Newman-Keuls post hoc multiple comparisons indicated statistically significant differences between V and P treatment (#, P < 0.05) and between V and EB+P (*, P < 0.01). The CaMKII total activity due to EB+P, compared with vehicle, was significant (#, P < 0.05) (n = 6 for each group). B, The homogenates were subjected to Western immunoblot analysis as described. Each lane represents the sample from one animal per treatment group. A representative autoradiograph is shown. The GAPDH band at 37 kDa was used as a normalizing control. Lane 1 corresponds to vehicle control (V); lane 2, EB; lane 3, P; and lane 4, EB+P treatments. Magic marker molecular weight (MW) standards were run alongside the protein samples and the MWs are noted adjacent to lane 1.