Mechanosensory and Chemosensory Primary Cilia in Ciliopathy and Ciliotherapy

Nauli, Surya M.; Sherpa, Rinzhin T.; Reese, Caretta J.; Nauli, Andromeda M.

doi:10.1002/9781118966174.ch5

Cited by 4 publications

(10 citation statements)

References 197 publications

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“…Despite these differences in conductance, channels at the tip and the base of the cilia as well as outside of cilia were blocked by Gd 3+ and activated by ATP in agreement with previous reports [1]. Heterogeneous distribution of channels throughout the membrane of cilia has been previously suggested based on either immunoflurescent staining or channel recordings in larger olfactory cilia [2,3,[13][14][15][16][17][18], however direct experimental evidence in smaller cilia as reported here was missing. The importance of using direct recording to prove functionality of channels has been recently demonstrated in a study reporting that PC2 forms functioning ion channel in primary cilia of kidney cells but has no measurable constitutive activity in the plasma membrane, which, as the authors themselves note, could not be ruled out based on fluorescence study only [19].…”

Section: Discussionsupporting

confidence: 91%

“…It displayed significantly different mean slope conductance and lacked the strong sensitivity to displacement. This lends further experimental support to previous observations and hypotheses that the population of channels in primary cilia is different to that of the cytoplasmic membrane [2,3,[13][14][15][16]18]. Furthermore, the markedly different character of the response to nanopipette displacement in the extra-ciliary membrane, together with Gd 3+ sensitivity of the response observed in cilia base indicates these changes are unlikely to be caused simply by deterioration of the seal between the nanopipette during the displacements and represent genuine changes in channel activity.…”

Section: Discussionsupporting

confidence: 87%

“…The primary cilium is a single immotile antenna-like organelle protruding from the membrane of most mammalian cells that comprises a distinct cellular compartment with a specific population of ion channels and receptors [1][2][3][4]. Although initially considered as an evolutionary vestigial structure, the primary cilium has been suggested to play a role in a number of physiological processes [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

“…Primary cilia display specific ultrastructure [7,10,11] and play a key role in the cellular response to chemical and mechanical alterations, serving as sensory platforms to flow, compression and pressure [2,5,6,12]. Various proteins such as polycystin-1 (PC1), polycystin-2 (PC2), transient receptor potential vanilloid channel 4 (TRPV4) or Piezo channels 1 and 2 have been shown to localise within the ciliary membrane and are postulated as signal transducers sensitive to mechanical alterations [2,3,[13][14][15][16][17][18][19][20]. In kidney epithelial cells, bending of the cilium due to fluid flow was shown to result in an intracellular increase in calcium mediated by the extracellular levels of calcium itself [13,14], while chondrocyte cilia respond to compression through a purinergic calcium signalling pathway [21,22].…”

Section: Introductionmentioning

confidence: 99%

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Nanoscale Mapping Reveals Functional Differences in Ion Channels Populating the Membrane of Primary Cilia

Torres-Pérez

Naeem

Thompson

et al. 2019

Cell Physiol Biochem

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Background/Aims: The primary cilium is a nanoscale membrane protrusion believed to act as a mechano-chemical sensor in a range of different cell types. Disruptions in its structure and signalling have been linked to a number of medical conditions, referred to as ciliopathies, but remain poorly understood due to lack of techniques capable of investigating signal transduction in cilia at nanoscale. Here we set out to use latest advances in nanopipette technology to address the question of ion channel distribution along the structure of primary cilium. Methods: We used glass nanopipettes and Scanning Ion Conductance Microscopy (SICM) to image 3D topography of intact primary cilia in inner medullary collecting duct (IMCD) cells with nanoscale resolution. The high-resolution topographical images were then used to navigate the nanopipette along the structure of each cilium and perform spatially resolved single-channel recordings under precisely controlled mechanical and chemical stimulation. Results:We have successfully obtained first single-channel recordings at specific locations of intact primary cilia. Our experiments revealed significant differences between the populations of channels present at the ciliary base, tip and within extra-ciliary regions in terms of mean conductance and sensitivity to membrane displacement as small as 100 nm. Ion channels at the base of cilium, where mechanical strain is expected to be the highest, appeared particularly sensitive to the mechanical displacement. Conclusion: Our results suggest the distribution of ion channels in the membrane of primary cilia is non-homogeneous. The relationship between the location and function of ciliary ion channels could be key to understanding signal transduction in primary cilia.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Nanoscale Mapping Reveals Functional Differences in Ion Channels Populating the Membrane of Primary Cilia

Torres-Pérez

Naeem

Thompson

et al. 2019

Cell Physiol Biochem

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“…For endothelial cell lines, we used previously generated mouse endothelial cells. 34 Prior to all experiments, cells at 75-85% confluence were differentiated for 24-48 h in serum-free media so we could accurately quantify and study the effects of CT-NPs in each experiment.…”

Section: Cell Culturementioning

confidence: 99%

Personalized Nanotherapy by Specifically Targeting Cell Organelles To Improve Vascular Hypertension

et al. 2018

Self Cite

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Ciliopathies caused by abnormal function of primary cilia include expanding spectrum of kidney, liver, and cardiovascular disorders. There is currently no treatment available for patients with cilia dysfunction. Therefore, we generated and compared two different (metal and polymer) ciliatargeted nanoparticle drug delivery systems (CTNDDS), CT-DAu-NPs and CT-PLGA-NPs, for the first time. These CTNDDS loaded with fenoldopam were further compared to fenoldopam-alone. Live-imaging of single-cell-single-cilium analysis confirmed that CTNDDS specifically targeted to primary cilia. While CTNDDS did not show any advantages over fenoldopam-alone in cultured cells in vitro, CTNDDS delivered fenoldopam more superior than fenoldopam-alone by eliminating the side effect of reflex tachycardia in murine models. Although slow infusion was required for fenoldopam-alone in mice, bolus injection was possible for CTNDDS. Though there were no significant therapeutic differences between CT-DAu-NPs and CT-PLGA-NPs, CT-PLGA-NPs tended to correct ciliopathy parameters closer to normal physiological levels, indicating CT-*

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MELD-Path Efficiently Computes Conformational Transitions, Including Multiple and Diverse Paths

Pérez

Sittel

Stock

et al. 2018

J. Chem. Theory Comput.

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The molecular actions of proteins occur along reaction coordinates. Current computer methods have limited ability to explore them. We describe a fast protocol called MELD-path that (1) efficiently samples relevant conformational states via MELD, an accelerator of Molecular Dynamics (MD), (2) seeds multiple short MD trajectories from MELD states, and then (3) constructs Markov State Models (MSM) that give the routes and kinetics. We tested the method against extensive (multi μs) MD simulations of the right-handed- to left-handed-helix transition of a 9-mer peptide of AIB, the symmetry of which allows us to establish convergence. MELD-path finds all the metastable states, their correct relative populations, and the full ensemble of routes, not just a single assumed route. For this transition, we find a very broad route structure. MELD-path is highly parallelizable and efficient, yielding the full route map in a few days of computation. We believe MELD-path could be a general and rapid way to explore mechanistic processes in biomolecules on the computer.

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Mechanosensory and Chemosensory Primary Cilia in Ciliopathy and Ciliotherapy

Cited by 4 publications

References 197 publications

Nanoscale Mapping Reveals Functional Differences in Ion Channels Populating the Membrane of Primary Cilia

Nanoscale Mapping Reveals Functional Differences in Ion Channels Populating the Membrane of Primary Cilia

Personalized Nanotherapy by Specifically Targeting Cell Organelles To Improve Vascular Hypertension

MELD-Path Efficiently Computes Conformational Transitions, Including Multiple and Diverse Paths

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