Di Benedetto, Giuseppe; (1991) Human respiratory epithelium: control of ciliary activity and techniques of investigation. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

In man, most of the upper airways and the tracheo-bronchial tree down to the non alveolar walls of the respiratory bronchioles are covered by ciliated epithelium. Mucociliary clearance is the most important clearing mechanism of the respiratory tract and is the result of beating cilia propelling the overlying secretions, carrying both trapped inhaled material and locally produced biological debris, toward the oropharynx. The rate of mucus transport is determined by the power produced by each cilium and the number of cilia in contact with the mucus. Both of these parameters are a function of ciliary beat frequency (CBF). This thesis deals with a technique for measuring CBF in vitro and how CBF is regulated in human respiratory cells. We measured CBF with a photometric technique and used a perfusional apparatus to observe changes in CBF in response to modifications of the extra- and intracellular microenvironments. We investigated the variability of CBF in human respiratory cells and assessed the reliability of a technique for storing ciliated cells at sub-zero temperatures, without affecting the basal CBF and the ciliary response to pharmacological stimulation. Our apparatus was tested in a study on the effect of Amiloride on ciliary activity, where we demonstrated a cilio-stimulatory action of the drug. The last two Chapters describe experiments on the role of cyclic AMP and calcium in regulating CBF in human respiratory epithelium. We assessed dose-response and time-response curves and used inhibitors of cyclic nucleotide- and calcium-dependent kinases to further clarify the intra-cellular pathways. The results indicate that in man, as in other species, both cyclic AMP and calcium regulate CBF.

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