Search Results (3)

Single cilia, 100 nm in diameter and 10 µm in length, were isolated from mouse tracheae with Triton X-100 (0.02%) treatment, and the effects of pH on ciliary beating were examined by measuring the ciliary beat frequency (CBF) and the ciliary bend distance (CBD—an index of amplitude) using a high-speed video microscope (250 fps). ATP (2.5 mM) plus 8Br-cAMP (10 µM) reactivated the CBF and CBD in the isolated cilia, similar to the cilia of in vivo tracheae. In the reactivated isolated cilia, an elevation in pH from 7.0 to 8.0 increased the CBF from 3 to 15 Hz and the CBD from 0.6 to 1.5 µm. The pH elevation also increased the velocity of the effective stroke; however, it did not increase the recovery stroke, and, moreover, it decreased the intervals between beats. This indicates that H⁺ (pH_i) directly acts on the axonemal machinery to regulate CBF and CBD. In isolated cilia priorly treated with 1 µM PKI-amide (a PKA inhibitor), 8Br-cAMP did not increase the CBF or CBD in the ATP-stimulated isolated cilia. pH modulates the PKA signal, which enhances the axonemal beating generated by the ATP-activated inner and outer dyneins. Full article

Ambroxol (ABX), a frequently prescribed secretolytic agent which enhances the ciliary beat frequency (CBF) and ciliary bend angle (CBA, an index of amplitude) by 30%, activates a voltage-dependent Ca²⁺ channel (Ca_V1.2) and a small transient Ca²⁺ release in the ciliated lung airway epithelial cells (c-LAECs) of mice. The activation of Ca_V1.2 alone enhanced the CBF and CBA by 20%, mediated by a pH_i increase_i and a [Cl⁻]_i decrease in the c-LAECs. The increase in pH_i, which was induced by the activation of the Na⁺-HCO₃⁻ cotransporter (NBC), enhanced the CBF (by 30%) and CBA (by 15–20%), and a decrease in [Cl⁻]_i, which was induced by the Cl⁻ release via anoctamine 1 (ANO1), enhanced the CBA (by 10–15%). While a Ca²⁺-free solution or nifedipine (an inhibitor of Ca_V1.2) inhibited 70% of the CBF and CBA enhancement using ABX, Ca_V1.2 enhanced most of the CBF and CBA increases using ABX. The activation of the Ca_V1.2 existing in the cilia stimulates the NBC to increase pH_i and ANO1 to decrease the [Cl⁻]_i in the c-LAECs. In conclusion, the pH_i increase and the [Cl⁻]_i decrease enhanced the CBF and CBA in the ABX-stimulated c-LAECs. Full article

In Ts1Rhr, a Down syndrome model mouse, the airway ciliary beatings are impaired; that is, decreases in ciliary beat frequency (CBF) and ciliary bend angle (CBA, an index of ciliary beat amplitude)). A resumption to two copies of the Pcp4 gene on the Ts1Rhr trisomic segment (Ts1Rhr:Pcp4^+/+/-) rescues the decreases in CBF and CBA that occur in Ts1Rhr. In airway cilia, upon stimulation with procaterol (a β₂-agonist), the CBF increase is slower over the time course than the CBA increase because of cAMP degradation by Ca²⁺/calmodulin-dependent phosphodiesterase 1 (PDE1) existing in the metabolon regulating CBF. In Ts1Rhr, procaterol-stimulated CBF increase was much slower over the time course than in the wild-type mouse (Wt) or Ts1Rhr:Pcp4^+/+/-. However, in the presence of 8MmIBMX (8-methoxymethyl isobutylmethyl xanthine, an inhibitor of PDE1) or calmidazolium (an inhibitor of calmodulin), in both Wt and Ts1Rhr, procaterol stimulates CBF and CBA increases over a similar time course. Measurements of cAMP revealed that the cAMP contents were lower in Ts1Rhr than in Wt or in Ts1Rhr:Pcp4^+/+/-, suggesting the activation of PDE1A that is present in Ts1Rhr airway cilia. Measurements of the intracellular Ca²⁺ concentration ([Ca²⁺]_i) in airway ciliary cells revealed that temperature (increasing from 25 to 37 °C) or 4αPDD (a selective transient receptor potential vanilloid 4 (TRPV4) agonist) stimulates a larger [Ca²⁺]_i increase in Ts1Rhr than in Wt or Ts1Rhr:Pcp4^+/+/-. In airway ciliary cells of Ts1Rhr, Pcp4-dose dependent activation of TRPV4 appears to induce an increase in the basal [Ca²⁺]_i. In early embryonic day mice, a basal [Ca²⁺]_i increased by PCP4 expressed may affect axonemal regulatory complexes regulated by the Ca²⁺-signal in Ts1Rhr, leading to a decrease in the basal CBF and CBA of airway cilia. Full article