Previously, we have reported that apoptosis of cerebellar granular neurons induced by incubation in 5 mM K+ and serum-free medium (LK-S) was associated with an increase in the delayed rectifier K+ current (IK). Here, we show that IK associated with apoptotic neurons is mainly encoded by a Kv2.1 subunit. Silencing Kv2.1 expression by small interfering RNA reduces IK and increases neuron viability. Forskolin is able to decrease the IK amplitude recording from neurons of both the LK-S and control group, and prevents apoptosis of granule cells that are induced by LK-S. Dibutyryl cAMP mimicks the effect of forskolin on the modulation of IK and, accordingly, the inhibitor of protein kinase A, H-89, aborts the neuron-protective effect induced by forskolin. Whereas the expression of Kv2.1 was silenced by Kv2.1 small interfering RNA, the inhibition of forskolin on the current amplitude was significantly reduced. Quantitative RT-PCR and whole-cell recording revealed that the expression of Kv2.1 was elevated in the apoptotic neurons, and forskolin significantly depressed the expression of Kv2.1. We conclude that the protection against apoptosis via the protein kinase A pathway is associated with a double modulation on IK channel properties and its expression of α-subunit that is mainly encoded by the Kv2.1 gene.