Protein Kinase C Regulates the Flow Rate-Dependent Decline in Human Nasal Ciliary Beat Frequency In Vitro

Abstract

Cilia provide the driving force for mucociliary clearance, the process that removes mucus from the airways. Protein kinase C (PKC) plays a poorly understood regulatory role in phosphorylation-based signal transduction cascades, including the control of human mucociliary clearance, especially with respect to ciliary beat frequency (CBF). Ciliary studies minimize the importance of fluid flow, because it is generally accepted that flow increases CBF. Here, we studied postflow events by measuring CBF in vitro in volunteers. Rose chamber–loaded cells were pulsed for 5 minutes at 30 mL/h in medium-199 ± PKC modulators at 20°C. The 5-minute pulse precipitated a fall in CBF noted within 1 minute after flow (acute dip response [ADR] to 84 ± 2% of preflow baseline). Thereafter, CBF rose to 8% below baseline for 30 minutes [postrecovery plateau at 92 ± 3%]. Preincubation with 1 μM of phorbol 12-myristate 13-acetate (PMA), a PKC-activating phorbol ester attenuated the ADR (c. 95%) and restored the postrecovery plateau almost to baseline levels (98 ± 0.7%; p > 0.10 compared with baseline CBF). With respect to the ADR, the PMA protective effect was lost in the presence of the selective PKC inhibitor myristoylated epidermal growth factor peptide 651–658 (Myr-PKCI; 10 μM). Myr-PKCI alone changed the ADR pattern such that the CBF remained at 15% below preflow baseline. We conclude that CBF fall and recovery after a fluid pulse is regulated by PKC activity either directly or indirectly.