Three types of ependymal cells with intracellular calcium oscillation are characterized by distinct cilia beating properties

Liu, Tongyu; Jin, Xingjian; Prasad, Rahul; Sari, Youssef; Nauli, Surya M.

doi:10.1002/jnr.23405

Cited by 16 publications

(21 citation statements)

References 29 publications

(38 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Moreover, we demonstrated that Ethanol at 0.25% concentration significantly repressed cilia beating frequency irrespective of their type (Figure 3). More importantly, these data are in consistence with our previous findings 10 .…”

Section: Representative Resultssupporting

confidence: 79%

“…Moreover, ciliary beating frequency should be measured at different locations along each ependymal section. We have previously demonstrated that ciliary beat frequency data generated using this technique has been consistently reproducible among 87 experimental observations 10 . Hence, ependymal cells are accurately classified depending on their ciliary beating.…”

Section: Discussionmentioning

confidence: 99%

“…For example, using live fluorescence imaging, it has been demonstrated that irrespective of the ependymal cell/cilia type, ependymal cells are characterized by unique calcium oscillation properties 10 . All in all, this protocol is relevant to both basic scientific knowledge and clinical practice.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Omran

Saternos

Liu

et al. 2015

JoVE

Self Cite

View full text Add to dashboard Cite

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various neurological deficits. The current ex vivo live imaging of motile ependymal cilia technique allows for a detailed study of ciliary dynamics following several steps. These steps include: mice euthanasia with carbon dioxide according to protocols of The University of Toledo's Institutional Animal Care and Use Committee (IACUC); craniectomy followed by brain removal and sagittal brain dissection with a vibratome or sharp blade to obtain very thin sections through the brain lateral ventricles, where the ependymal cilia can be visualized. Incubation of the brain's slices in a customized glass-bottom plate containing Dulbecco's Modified Eagle's Medium (DMEM)/High-Glucose at 37 °C in the presence of 95%/5% O 2 /CO 2 mixture is essential to keep the tissue alive during the experiment. A video of the cilia beating is then recorded using a high-resolution differential interference contrast microscope. The video is then analyzed frame by frame to calculate the ciliary beating frequency. This allows distinct classification of the ependymal cells into three categories or types based on their ciliary beating frequency and angle. Furthermore, this technique allows the use of high-speed fluorescence imaging analysis to characterize the unique intracellular calcium oscillation properties of ependymal cells as well as the effect of pharmacological agents on the calcium oscillations and the ciliary beating frequency. In addition, this technique is suitable for immunofluorescence imaging for ciliary structure and ciliary protein localization studies. This is particularly important in disease diagnosis and phenotype studies. The main limitation of the technique is attributed to the decrease in live motile cilia movement as the brain tissue starts to die. Video LinkThe video component of this article can be found at

show abstract

Section: Representative Resultssupporting

confidence: 79%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Omran

Saternos

Liu

et al. 2015

JoVE

Self Cite

View full text Add to dashboard Cite

show abstract

“…The combinatorial powers of ciliary parameters described in our study, such as beat direction, kinematics, and coordination, suggest a richness of potential scenarios for shaping the extracellular fluid environment at multiple scales to drive tissue homeostasis and remodeling, much as described for other tissue-organizing mechanical forces (44). Indeed, new imaging techniques have recently mapped both structurally distinct populations of cilia and spatiotemporally varying ciliary flow dynamics, in the ventricles of the mouse brain (45,46). Multiscale analyses of ciliated tissues not only reveal new tissue-level phenomena, but also enable a quantification of their functional roles in different tissue environments.…”

Section: Discussionmentioning

confidence: 75%

Motile cilia create fluid-mechanical microhabitats for the active recruitment of the host microbiome

Nawroth

Guo

Koch

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

106

103

View full text Add to dashboard Cite

We show that mucociliary membranes of animal epithelia can create fluid-mechanical microenvironments for the active recruitment of the specific microbiome of the host. In terrestrial vertebrates, these tissues are typically colonized by complex consortia and are inaccessible to observation. Such tissues can be directly examined in aquatic animals, providing valuable opportunities for the analysis of mucociliary activity in relation to bacteria recruitment. Using the squid-vibrio model system, we provide a characterization of the initial engagement of microbial symbionts along ciliated tissues. Specifically, we developed an empirical and theoretical framework to conduct a census of ciliated cell types, create structural maps, and resolve the spatiotemporal flow dynamics. Our multiscale analyses revealed two distinct, highly organized populations of cilia on the host tissues. An array of long cilia (∼25 µm) with metachronal beat creates a flow that focuses bacteria-sized particles, at the exclusion of larger particles, into sheltered zones; there, a field of randomly beating short cilia (∼10 µm) mixes the local fluid environment, which contains host biochemical signals known to prime symbionts for colonization. This cilia-mediated process represents a previously unrecognized mechanism for symbiont recruitment. Each mucociliary surface that recruits a microbiome such as the case described here is likely to have system-specific features. However, all mucociliary surfaces are subject to the same physical and biological constraints that are imposed by the fluid environment and the evolutionary conserved structure of cilia. As such, our study promises to provide insight into universal mechanisms that drive the recruitment of symbiotic partners.cilia | microfluidics | host-bacterial symbiosis | biological fluid mechanics, biofiltration M any eukaryotic cells feature motile cilia, microtubulebased surface actuators that sense and propel the extracellular fluidic environment (1-3). Whereas cilia and cilia-like structures that sort and capture bacteria or particles are common and well-characterized features of aquatic organisms (4-7), in terrestrial animals such as mammals, the internal location of ciliated surfaces has made them difficult to study. A central challenge in internal ciliated mucus membranes, such as those lining the fallopian tube, the Eustachian tube, and the respiratory system (8), is to reconcile the effective clearance of toxic molecules and undesirable microbes with selective engagement of beneficial symbionts. For example, on airway epithelia, the coordinated beat of motile cilia creates a horizontal flow across their tips (9-12), which clears mucus, microorganisms, and debris (Fig. 1A). Disruption of this mucociliary clearance can lead to chronic infection of the airways (13). However, this simple model is incomplete; ciliated airway epithelia not only serve a clearance function, but also provide a habitat and a gateway for coevolved symbionts that play an essential role in the development of the host...

show abstract

“…Thus, we next investigated whether the hydrocephalus in the SrGAP3 −/− mice might be related to ciliary defects. The ependymal cells lining the ventricles in the adult brain are multiciliated epithelial cells and abnormal ependymal cilia result in hydrocephalus induced by anomalous flow of the CSF (Liu et al 2014). We showed that the length of the cilia was not altered in these mice but that the density of these cilia was significantly reduced.…”

Section: Discussionmentioning

confidence: 87%

SrGAP3 knockout mice display enlarged lateral ventricles and specific cilia disturbances of ependymal cells in the third ventricle

et al. 2015

View full text Add to dashboard Cite

In several mouse models of mental retardation, ventricular enlargements have been observed. Mutation in the SrGAP3 gene residing on chromosome 3p25 has previously been associated with intellectual disability in humans. In addition, SrGAP3 is related to Rho-GAPs signaling pathways, which play essential roles in the development and plasticity of the nervous system. About 10 % of postnatal homozygous SrGAP3-deficient mice die due to hydrocephalus, whereas the remaining mice survive into adulthood but display enlarged ventricles. We analyze the ventricular enlargement of these mice by performing a post-mortem MRI approach. We found a more than 15-fold enlargement of the lateral ventricles of homozygous SrGAP3-deficient mice. Moreover, we demonstrate that this phenotype was not accompanied by a stenosis of the aqueduct. Instead, SrGAP3 knockout mice displayed reduced densities of cilia of ependymal cells in These third ventricle compared to age-matched controls. This results indicate that the ventricular enlargement may be due to ciliopathy.

show abstract

Three types of ependymal cells with intracellular calcium oscillation are characterized by distinct cilia beating properties

Cited by 16 publications

References 29 publications

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Motile cilia create fluid-mechanical microhabitats for the active recruitment of the host microbiome

SrGAP3 knockout mice display enlarged lateral ventricles and specific cilia disturbances of ependymal cells in the third ventricle

Contact Info

Product

Resources

About