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05). Seventy percent (12 out of 17 cells) of DBB neurons responded to PrP(106�C126). The PrP(106�C126) response was time independent and did not desensitize on subsequent applications. ��Scrambled�� PrP(106�C126) peptide with a different order of the same amino acid constituents (inactive control) had no effect on whole-cell currents (control = 8.37 �� 0.65 nA, ��scrambled PrP�� = 8.33 �� 0.69 nA; P > 0.05; Fig.1B). Outward potassium currents recorded from DDB neurons in the voltage range �C30 mV to +30 mV are a mixture of calcium-activated and noncalcium-activated components (Jhamandas et al., 2001). The noncalcium-activated component consists primarily of the delayed rectifier (IK) and the transient outward (IA) currents. The calcium-dependent component of potassium currents includes GUCY1B3 voltage-sensitive conductances, IC (BK channels). BKM120 chemical structure The effects of PrP(106�C126) on these conductances were further investigated. IK and IA potassium currents are voltage-sensitive currents. To activate IA, the cells are hyperpolarized to �C120 mV from a holding potential of �C80 mV for the removal of inactivation. IA is inactivated at �C40 mV, but this does not affect IK currents. Using these differences in biophysical properties, we are able to isolate and investigate the effects of PrP(106�C126) on these voltage-sensitive potassium currents. Hence, a conditioning pulse to �C40 mV will activate IK only (Connor and Stevens, 1971; Jhamandas et al., 2001), whereas a conditioning pulse to �C120 mV will activate both IA and IK. Subtracting the currents from conditioning pulse of �C40 mV from that of �C120 mV will isolate the IA current. Figure check details 2 shows currents recorded from a neuron with a conditioning pulse of �C40 mV for 150 msec in control (Fig.2A), presence of PrP(106�C126) (Fig. 2B), and washout (Fig. 2C). The I-V graph (Fig. 2D) shows a significant decrease of 14.8% in the presence of PrP(106�C126) (control = 9.88 �� 0.54nA, PrP(106�C126) = 8.42 �� 0.54 nA, n = 18; *P
