A family of accessory beta subunits significantly contributes to the functional diversity of large-conductance, Ca(2+)- and voltage-dependent potassium (BK) channels in native cells. Here we describe the functional properties of one variant of the beta subunit family, which confers properties on BK channels totally unlike any that have as yet been observed. Coexpression of this subunit (termed beta3) with Slo alpha subunits results in rectifying outward currents and, at more positive potentials, rapidly inactivating ( approximately 1 msec) currents. The underlying rapid inactivation process results in an increase in the apparent activation rate of macroscopic currents, which is coupled with a shift in the activation range of the currents at low Ca(2+). As a consequence, the currents exhibit more rapid activation at low Ca(2+) relative to any other BK channel subunit combinations that have been examined. In part because of the rapid inactivation process, single channel openings are exceedingly brief. Although variance analysis suggests a conductance in excess of 160 pS, fully resolved single channel openings are not observed. The inactivation process results from a cytosolic N-terminal domain of the beta3 subunit, whereas an extended C-terminal domain does not participate in the inactivation process. Thus, the beta3 subunit appears to use a rapid inactivation mechanism to produce a current with a relatively rapid apparent activation time course at low Ca(2+). The beta3 subunit is a compelling example of how the beta subunit family can finely tune the gating properties of Ca(2+)- and voltage-dependent BK channels.

The hyperpolarization-activated cation channels (Ih) play a distinct role in rhythmic activities in a variety of tissues, including neurons and cardiac cells. In the present study, we investigated whether Ca 2؉ can permeate through the hyperpolarization-activated pacemaker channels (HCN) expressed in HEK293 cells and Ih channels in dorsal root ganglion (DRG) neurons. Using combined measurements of whole-cell currents and fura-2 Ca 2؉ imaging, we found that there is a Ca 2؉ influx in proportion to Ih induced by hyperpolarization in HEK293 cells. The Ih channel blockers Cs ؉ and ZD7288 inhibit both HCN current and Ca 2؉ influx. Measurements of the fractional Ca 2؉ current showed that it constitutes 0.60 ؎ 0.02% of the net inward current through HCN4 at ؊120 mV. This fractional current is similar to that of the low Ca 2؉ -permeable AMPA-R (␣-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor) channels in Purkinje neurons. In DRG neurons, activation of Ih for 30 s also resulted in a Ca 2؉ influx and an elevated action potentialinduced secretion, as assayed by the increase in membrane capacitance. These results suggest a functional significance for Ih channels in modulating neuronal secretion by permitting Ca 2؉ influx at negative membrane potentials.

Abeta accumulation has an important function in the etiology of Alzheimer's disease (AD) with its typical clinical symptoms, like memory impairment and changes in personality. However, the mode of this toxic activity is still a matter of scientific debate. We used the APP/PS1KI mouse model for AD, because it is the only model so far which develops 50% hippocampal CA1 neuron loss at the age of 1 year. Previously, we have shown that this model develops severe learning deficits occurring much earlier at the age of 6 months. This observation prompted us to study the anatomical and cellular basis at this time point in more detail. In the current report, we observed that at 6 months of age there is already a 33% CA1 neuron loss and an 18% atrophy of the hippocampus, together with a drastic reduction of long-term potentiation and disrupted paired pulse facilitation. Interestingly, at 4 months of age, there was no long-term potentiation deficit in CA1. This was accompanied by reduced levels of pre-and post-synaptic markers. We also observed that intraneuronal and total amount of different Abeta peptides including N-modified, fibrillar and oligomeric Abeta species increased and coincided well with CA1 neuron loss. Overall, these data provide the basis for the observed robust working memory deficits in this mouse model for AD at 6 months of age.

Action potentials (APs) are the principal physiological stimuli for neurotransmitter secretion in neurons. Most studies on stimulus-secretion coupling have been performed under voltage clamp using artificial electrical stimuli. To investigate the modulatory effects of AP codes on neural secretion, we introduce a capacitance method to study AP-induced secretion in single cells. The action potential pattern was defined by a four-parameter "code function:" F(n, m, f, d). With this method, cell secretion evoked by stimulation with an AP code was quantified in real time by membrane capacitance (Cm) in adrenal chromaffin cells. We found, in addition to AP frequency (f), for a given number of APs, another parameter of the AP code, the number of AP bursts (m) in which the set of APs occurs, can effectively modulate cell secretion. Possible mechanisms of the m effect are depletion of the readily releasable pool and inactivation of Ca2+ channels during a burst of APs. The physiological m effect may play a key role in AP-mediated neural information transfer within a single neuron and among the elements of a neural network.