An experimental and computational analysis of primary cilia deflection under fluid flow

In this study we have developed a novel model of the deflection of primary cilia experiencing fluid flow accounting for phenomena not previously considered. Specifically, we developed a large rotation formulation that accounts for rotation at the base of the cilium, the initial shape of the cilium and fluid drag at high deflection angles. We utilised this model to analyse full 3D data-sets of primary cilia deflecting under fluid flow acquired with high-speed confocal microscopy. We found a wide variety of previously unreported bending shapes and behaviours. We also analysed post-flow relaxation patterns. Results from our combined experimental and theoretical approach suggest that the average flexural rigidity of primary cilia might be higher than previously reported (Schwartz et al. 1997, Am J Physiol. 272(1 Pt 2):F132–F138). In addition our findings indicate that the mechanics of primary cilia are richly varied and mechanisms may exist to alter their mechanical behaviour.     

A shear-rate-dependent flow generated via magnetically controlled metachronal motion of artificial cilia

Biomechanics and Modeling in Mechanobiology - Cilia beating is a naturally occurring phenomenon that can be utilized in fluid transport in designing several biomechanical devices. Inspired by the...     

Spontaneous creation of macroscopic flow and metachronal waves in an array of cilia

Cells carrying cilia on their surface show many striking features: alignment of cilia in an array, two-phase asymmetric beating for each cilium, and existence of metachronal coordination with a constant phase difference between two adjacent cilia. We give simple theoretical arguments based on hydrodynamic coupling and an internal mechanism of the cilium derived from the behavior of a collection of molecular motors to account qualitatively for these cooperative features. Hydrodynamic interactions can lead to the alignment of an array of cilia. We study the effect of a transverse external flow and obtain a two-phase asymmetrical beating, faster along the flow and slower against the flow, proceeding around an average curved position. We show that an aligned array of cilia is able to spontaneously break the left-right symmetry and to create a global average flow. Metachronal coordination arises as a consequence of the internal mechanism of the cilia and their hydrodynamic couplings, with a wavelength comparable to that found in experiments. It allows the cilia to start beating at a lower adenosine-triphosphate threshold and at a higher frequency than for a single cilium. It also leads to a rather stationary flow, which might be its major advantage.     

Rotation and translocation of microtubules in vitro induced by dyneins from Tetrahymena cilia

Dynein, the force-generating enzyme that powers the movement of cilia and flagella, has been characterized biochemically, but no simple system has been available for examining its motile properties. Here we describe a quantitative in vitro motility assay in which dynein adsorbed onto a glass surface induces linear translocation of purified bovine microtubules. Using this assay, we show that both 22S and 14S dyneins from Tetrahymena cilia induce movement but have distinct motile properties. A unique property of 14S dynein, which has not been described for other motility proteins, is its ability to generate torque that causes microtubules to rotate during forward translocation. In the axoneme, 14S dynein-induced torque may induce rotation of central-pair microtubules and may play an important role in generating three-dimensional ciliary beating patterns.     

Primary cilia of inv/inv mouse renal epithelial cells sense physiological fluid flow: bending of primary cilia and Ca2+ influx

Primary cilia are hypothesized to act as a mechanical sensor to detect renal tubular fluid flow. Anomalous structure of primary cilia and/or impairment of increases in intracellular Ca2+ concentration in response to fluid flow are thought to result in renal cyst formation in conditional kif3a knockout, Tg737 and pkd1/pkd2 mutant mice. The mutant inv/inv mouse develops multiple renal cysts like kif3a, Tg737 and pkd1/pkd2 mutants. Inv proteins have been shown to be localized in the renal primary cilia, but response of inv/inv cilia to fluid stress has not been examined. In the present study, we examined the mechanical response of primary cilia to physiological fluid flow using a video microscope, as well as intracellular Ca2+ increases in renal epithelial cells from normal and inv/inv mice in response to flow stress. Percentages of ciliated cells and the length of primary cilia were not significantly different between primary renal cell cultures from normal and inv/inv mutant mice. Localization of inv protein was restricted to the base of primary cilia even under flow stress. Inv/inv mutant cells had similar bending mechanics of primary cilia in response to physiological fluid flow compared to normal cells. Furthermore, no difference was found in intracellular Ca2+ increases in response to physiological fluid flow between normal and inv/inv mutant cells. Our present study suggests that the function of the inv protein is distinct from polaris (the Tg737 gene product), polycystins (pkd1 and pkd2 gene products).     

Analytical ultrastructural investigation of microliths in salivary glands of cat

Summary Microliths in Araldite-embedded pieces of submandibular and sublingual glands of cat were stained in semithin sections by Methylene Blue and Azure II followed by Basic Fuchsin, and were examined in ultrathin sections by electronmicroscopical X-ray microanalysis. Calcium and phosphorus were detected in substantial aggregates of crystals that were stained by Basic Fuchsin and appeared to be hydroxyapatite, but were not detected in granular material that was stained by Methylene Blue and Azure II and appeared to be organic. The polychromatic stain thus appears to be a useful indicator of calcified material. The majority of microliths in acini contained substantial aggregates of crystals, whereas the majority of those in ducts did not. This corresponds to the distribution of the glandular calcium, and suggests that microliths are variously enriched with calcium according to its local level.     

The resorption of cilia

Conclusions Ciliary resorption, a phenomenon consistently occurring in certain protozoa, illustrates fundamental cellular processes. Membranes are observed to break and fuse in an end-to-end manner and to laminate by surface fusion within the environment of the resorption vesicle. The sequence and manner of filament degradation demonstrate the relative lability of central filaments as compared to peripheral ones and the tenacity of the arrangement of peripheral filaments.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his eightieth birthday.This study was supported by a grant (C-5581) from the National Cancer Institute, United States Public Health Service.     

An investigation into induced plant defences

This laboratory exercise involves techniques that can be used to investigate induced plant defences. Runner beans are damaged, and the leaves that subsequently grow are tested for the presence of additional defence mechanisms. A control (undamaged) group is kept for comparison. Induced physical defences are indicated by an increase in the density of hairs (trichomes) on the leaves, and induced chemical defences by a decrease in the palatability of the leaves. The latter is tested by a bioassay using garden snails. These basic methods can be used to investigate a variety of defensive responses in plants.     

Cinefilms of particle capture by an induced local change of beat of lateral cilia of a bryozoan

A local disruption of the metachronal wave always accompanies capture of algal cells by tentacles of Flustrellidra hispida (Fabricius). Beat changes for ≈0.2 s over ≈100μm of the ciliated band during capture of a 10-μm particle. The halted parcel of water is therefore larger than the particle of food but much smaller than the flow that continues past the tentacles elsewhere. These events are consistent with the hypothesis that an induced local reversal of beat concentrates particles for those suspension feeders that retain particles upstream from a band of simple cilia (adults or larvae of bryozoans, brachiopods, phoronids, hemichordates, and echinoderms). These events are not explained by other hypotheses that have been advanced for concentration of particles by these suspension feeders. Aerosol filtration models of direct interception are not applicable to this type of ciliary suspension feeder because retention depends on the magnitude of a stimulus and response to it. The stimulus will not be the same function of diameter of the food particle, and response is unlikely below a threshold stimulus.     

Ultrastructural abnormalities of respiratory cilia

Ultrastructural abnormalities of human respiratory tract cilia have been studied in 33 patients: 21 were adults, 18 with chronic bronchitis, and 12 were children, two with situs inversus. Abnormalities, such as the lack of a few dynein arms or the loss of a peripheral doublet were observed quite frequently in both children and adults. However, congenital abnormalities associated with the "immotile cilia syndrome" were rare and were observed in only three of the children and none of the adults.     

Planar polarity of ependymal cilia

Ependymal cells, epithelial cells that line the cerebral ventricles of the adult brain in various animals, extend multiple motile cilia from their apical surface into the ventricles. These cilia move rapidly, beating in a direction determined by the ependymal planar cell polarity (PCP). Ciliary dysfunction interferes with cerebrospinal fluid circulation and alters neuronal migration. In this review, we summarize recent studies on the cellular and molecular mechanisms underlying two distinct types of ependymal PCP. Ciliary beating in the direction of fluid flow is established by a combination of hydrodynamic forces and intracellular planar polarity signaling. The ciliary basal bodies' anterior position on the apical surface of the cell is determined in the embryonic radial glial cells, inherited by ependymal cells, and established by non-muscle myosin II in early postnatal development.     

Numerical investigation of mucociliary clearance using power law and thixotropic mucus layers under discrete and continuous cilia motion

Biomechanics and Modeling in Mechanobiology - Mucus layer movement inside the airway system is an important phenomenon as the first defensive mechanism against pathogens. This research deals with...     

Analytical investigation of styrax and benzoin balsams by HPLC- PAD-fluorimetry and GC-MS

An HPLC and GC study has been conducted on the aromatic oleoresins styrax and benzoin produced by several American, Mediterranean and East-Asian trees, and widely used in ancient civilisations for their therapeutic and odoriferous properties. Initial experiments were performed by HPLC-PAD-fluorimetry for the analysis of several aromatic components, and then completed by GC-MS for the characterisation of both aromatic and triterpenic derivatives. In this work, it was crucial to isolate from fresh natural exudates, and to characterise by two-dimensional NMR, some of the major constituents in order to extend the standard molecular pool prior to chromatographic identifications. This study reveals coniferyl benzoate as an excellent distinctive fluorescent biomarker of Siam benzoin substrate. It also confirms that fluorimetric-coupled detection is a powerful analytical tool for the identification of compounds in Hamamelidaceae extracts that are almost undetectable by UV. GC-MS was successfully applied to the determination of the botanical origin of Sumatra benzoin, and to the identification of lupeol [3beta-lup-20(29)-en-3-ol] for the first time in such balsam-type materials.