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Article ; Online: Nanostructure of bioactive glass affects bone cell attachment via protein restructuring upon adsorption.

Thamma, Ukrit / Kowal, Tia J / Falk, Matthias M / Jain, Himanshu

2021 Volume 11, Issue 1, Page(s) 5763

Abstract: The nanostructure of engineered bioscaffolds has a profound impact on cell response, yet its understanding remains incomplete as cells interact with a highly complex interfacial layer rather than the material itself. For bioactive glass scaffolds, this ... ...

Abstract	The nanostructure of engineered bioscaffolds has a profound impact on cell response, yet its understanding remains incomplete as cells interact with a highly complex interfacial layer rather than the material itself. For bioactive glass scaffolds, this layer comprises of silica gel, hydroxyapatite (HA)/carbonated hydroxyapatite (CHA), and absorbed proteins-all in varying micro/nano structure, composition, and concentration. Here, we examined the response of MC3T3-E1 pre-osteoblast cells to 30 mol% CaO-70 mol% SiO
MeSH term(s)	Adsorption ; Animals ; Carbonates/chemistry ; Cell Adhesion ; Cell Count ; Cell Line ; Cell Size ; Durapatite/chemistry ; Glass/chemistry ; Mice ; Nanopores ; Nanostructures/chemistry ; Nanostructures/ultrastructure ; Osteocytes/cytology ; Protein Structure, Secondary ; Proteins/chemistry ; Serum Albumin, Bovine/chemistry ; Spectroscopy, Fourier Transform Infrared
Chemical Substances	Carbonates ; Proteins ; Serum Albumin, Bovine (27432CM55Q) ; Durapatite (91D9GV0Z28)
Language	English
Publishing date	2021-03-11
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-85050-7
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Article ; Online: Compartmentalized ciliation changes of oligodendrocytes in aged mouse optic nerve.

Ning, Ke / Tran, Matthew / Kowal, Tia J / Mesentier-Louro, Louise A / Sendayen, Brent E / Wang, Qing / Lo, Chien-Hui / Li, Tingting / Majumder, Rishab / Luo, Jian / Hu, Yang / Liao, Yaping Joyce / Sun, Yang

Journal of neuroscience research

2024 Volume 102, Issue 1, Page(s) e25273

Abstract: Primary cilia are microtubule-based sensory organelles that project from the apical surface of most mammalian cells, including oligodendrocytes, which are myelinating cells of the central nervous system (CNS) that support critical axonal function. ... ...

Abstract	Primary cilia are microtubule-based sensory organelles that project from the apical surface of most mammalian cells, including oligodendrocytes, which are myelinating cells of the central nervous system (CNS) that support critical axonal function. Dysfunction of CNS glia is associated with aging-related white matter diseases and neurodegeneration, and ciliopathies are known to affect CNS white matter. To investigate age-related changes in ciliary profile, we examined ciliary length and frequency in the retinogeniculate pathway, a white matter tract commonly affected by diseases of aging but in which expression of cilia has not been characterized. We found expression of Arl13b, a marker of primary cilia, in a small group of Olig2-positive oligodendrocytes in the optic nerve, optic chiasm, and optic tract in young and aged C57BL/6 wild-type mice. While the ciliary length and ciliated oligodendrocyte cells were constant in young mice in the retinogeniculate pathway, there was a significant increase in ciliary length in the anterior optic nerve as compared to the aged animals. Morphometric analysis confirmed a specific increase in the ciliation rate of CC1
MeSH term(s)	Animals ; Mice ; Mice, Inbred C57BL ; Optic Nerve ; Oligodendroglia ; Neuroglia ; White Matter ; Mammals
Language	English
Publishing date	2024-01-27
Publishing country	United States
Document type	Journal Article
ZDB-ID	195324-2
ISSN	1097-4547 ; 0360-4012
ISSN (online)	1097-4547
ISSN	0360-4012
DOI	10.1002/jnr.25273
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Article: Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids.

Ning, Ke / Luo, Ziming / Kowal, Tia J / Tran, Matthew / Majumder, Rishab / Jarin, Trent M / Wu, Albert Y / Goldberg, Jeffrey L / Sun, Yang

Stem cells international

2023 Volume 2023, Page(s) 6494486

Abstract: Objectives: Primary cilia are conserved organelles found in polarized mammalian cells that regulate neuronal growth, migration, and differentiation. Proper cilia formation is essential during eye development. Our previous reports found that both ... ...

Abstract	Objectives: Primary cilia are conserved organelles found in polarized mammalian cells that regulate neuronal growth, migration, and differentiation. Proper cilia formation is essential during eye development. Our previous reports found that both amacrine and retinal ganglion cells (RGCs) contain primary cilia in primate and rodent retinas. However, whether primary cilia are present in the inner retina of human retinal organoids remains unknown. The purpose of this study is to characterize the primary cilia distribution in human embryonic stem cell (hESC-derived retinal organoid development. Materials and methods: Retinal organoids were differentiated from a hESC line, harvested at various developmental timepoints (day 44-day 266), and immunostained with antibodies for primary cilia, including Arl13b (for the axoneme), AC3, and Centrin3 (for the basal body). AP2 Results: A group of ciliated cells were present in the inner aspects of retinal organoids from day 44 to day 266 in culture. Ciliated Chx10-positive retinal progenitor cells, GFAP-positive astrocytes, and PKC Conclusions: Amacrine cells in retinal organoids retain cilia throughout development, whereas RGC ciliation gradually and progressively decreases with organoid maturation.
Language	English
Publishing date	2023-01-13
Publishing country	United States
Document type	Journal Article
ZDB-ID	2573856-2
ISSN	1687-9678 ; 1687-966X
ISSN (online)	1687-9678
ISSN	1687-966X
DOI	10.1155/2023/6494486
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Article ; Online: Nanostructure of bioactive glass affects bone cell attachment via protein restructuring upon adsorption

Ukrit Thamma / Tia J. Kowal / Matthias M. Falk / Himanshu Jain

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 14

Abstract: Abstract The nanostructure of engineered bioscaffolds has a profound impact on cell response, yet its understanding remains incomplete as cells interact with a highly complex interfacial layer rather than the material itself. For bioactive glass ... ...

Abstract	Abstract The nanostructure of engineered bioscaffolds has a profound impact on cell response, yet its understanding remains incomplete as cells interact with a highly complex interfacial layer rather than the material itself. For bioactive glass scaffolds, this layer comprises of silica gel, hydroxyapatite (HA)/carbonated hydroxyapatite (CHA), and absorbed proteins—all in varying micro/nano structure, composition, and concentration. Here, we examined the response of MC3T3-E1 pre-osteoblast cells to 30 mol% CaO–70 mol% SiO2 porous bioactive glass monoliths that differed only in nanopore size (6–44 nm) yet resulted in the formation of HA/CHA layers with significantly different microstructures. We report that cell response, as quantified by cell attachment and morphology, does not correlate with nanopore size, nor HA/CHO layer micro/nano morphology, or absorbed protein amount (bovine serum albumin, BSA), but with BSA’s secondary conformation as indicated by its β-sheet/α-helix ratio. Our results suggest that the β-sheet structure in BSA interacts electrostatically with the HA/CHA interfacial layer and activates the RGD sequence of absorbed adhesion proteins, such as fibronectin and vitronectin, thus significantly enhancing the attachment of cells. These findings provide new insight into the interaction of cells with the scaffolds’ interfacial layer, which is vital for the continued development of engineered tissue scaffolds.
Keywords	Medicine ; R ; Science ; Q
Subject code	500
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
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Article ; Online: Distribution of prototypical primary cilia markers in subtypes of retinal ganglion cells.

Kowal, Tia J / Dhande, Onkar S / Wang, Biao / Wang, Qing / Ning, Ke / Liu, Wendy / Berbari, Nicolas F / Hu, Yang / Sun, Yang

The Journal of comparative neurology

2022 Volume 530, Issue 12, Page(s) 2176–2187

Abstract: Loss of retinal ganglion cells (RGCs) underlies several forms of retinal disease including glaucomatous optic neuropathy, a leading cause of irreversible blindness. Several rare genetic disorders associated with cilia dysfunction have retinal ... ...

Abstract	Loss of retinal ganglion cells (RGCs) underlies several forms of retinal disease including glaucomatous optic neuropathy, a leading cause of irreversible blindness. Several rare genetic disorders associated with cilia dysfunction have retinal degeneration as a clinical hallmark. Much of the focus of ciliopathy associated blindness is on the connecting cilium of photoreceptors; however, RGCs also possess primary cilia. It is unclear what roles RGC cilia play, what proteins and signaling machinery localize to RGC cilia, or how RGC cilia are differentiated across the subtypes of RGCs. To better understand these questions, we assessed the presence or absence of a prototypical cilia marker Arl13b and a widely distributed neuronal cilia marker AC3 in different subtypes of mouse RGCs. Interestingly, not all RGC subtype cilia are the same and there are significant differences even among these standard cilia markers. Alpha-RGCs positive for osteopontin, calretinin, and SMI32 primarily possess AC3-positive cilia. Directionally selective RGCs that are CART positive or Trhr positive localize either Arl13b or AC3, respectively, in cilia. Intrinsically photosensitive RGCs differentially localize Arl13b and AC3 based on melanopsin expression. Taken together, we characterized the localization of gold standard cilia markers in different subtypes of RGCs and conclude that cilia within RGC subtypes may be differentially organized. Future studies aimed at understanding RGC cilia function will require a fundamental ability to observe the cilia across subtypes as their signaling protein composition is elucidated. A comprehensive understanding of RGC cilia may reveal opportunities to understanding how their dysfunction leads to retinal degeneration.
MeSH term(s)	Animals ; Biomarkers/metabolism ; Blindness ; Cilia ; Glaucoma/metabolism ; Mice ; Retinal Degeneration/etiology ; Retinal Ganglion Cells/metabolism
Chemical Substances	Biomarkers
Language	English
Publishing date	2022-04-17
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	3086-7
ISSN	1096-9861 ; 0021-9967 ; 0092-7317
ISSN (online)	1096-9861
ISSN	0021-9967 ; 0092-7317
DOI	10.1002/cne.25326
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Article ; Online: Cilia-associated wound repair mediated by IFT88 in retinal pigment epithelium.

Ning, Ke / Bhuckory, Mohajeet B / Lo, Chien-Hui / Sendayen, Brent E / Kowal, Tia J / Chen, Ming / Bansal, Ruchi / Chang, Kun-Che / Vollrath, Douglas / Berbari, Nicolas F / Mahajan, Vinit B / Hu, Yang / Sun, Yang

Scientific reports

2023 Volume 13, Issue 1, Page(s) 8205

Abstract: Primary cilia are conserved organelles that integrate extracellular cues into intracellular signals and are critical for diverse processes, including cellular development and repair responses. Deficits in ciliary function cause multisystemic human ... ...

Abstract	Primary cilia are conserved organelles that integrate extracellular cues into intracellular signals and are critical for diverse processes, including cellular development and repair responses. Deficits in ciliary function cause multisystemic human diseases known as ciliopathies. In the eye, atrophy of the retinal pigment epithelium (RPE) is a common feature of many ciliopathies. However, the roles of RPE cilia in vivo remain poorly understood. In this study, we first found that mouse RPE cells only transiently form primary cilia. We then examined the RPE in the mouse model of Bardet-Biedl Syndrome 4 (BBS4), a ciliopathy associated with retinal degeneration in humans, and found that ciliation in BBS4 mutant RPE cells is disrupted early during development. Next, using a laser-induced injury model in vivo, we found that primary cilia in RPE reassemble in response to laser injury during RPE wound healing and then rapidly disassemble after the repair is completed. Finally, we demonstrated that RPE-specific depletion of primary cilia in a conditional mouse model of cilia loss promoted wound healing and enhanced cell proliferation. In summary, our data suggest that RPE cilia contribute to both retinal development and repair and provide insights into potential therapeutic targets for more common RPE degenerative diseases.
MeSH term(s)	Mice ; Humans ; Animals ; Retinal Pigment Epithelium ; Cilia/physiology ; Retinal Degeneration ; Ciliopathies ; Disease Models, Animal ; Tumor Suppressor Proteins ; Microtubule-Associated Proteins
Chemical Substances	IFT88 protein, human ; Tumor Suppressor Proteins ; BBS4 protein, mouse ; Microtubule-Associated Proteins
Language	English
Publishing date	2023-05-21
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-35099-3
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Article ; Online: Primary cilia found on HeLa and other cancer cells.

Kowal, Tia J / Falk, Matthias M

Cell biology international

2015 Volume 39, Issue 11, Page(s) 1341–1347

Abstract: For many years now, researchers have known of a sensory appendage on the surface of most differentiated cell types called primary cilium. Primary cilia are both chemo- and mechano-sensory in function and have an obvious role in cell cycle control. ... ...

Abstract	For many years now, researchers have known of a sensory appendage on the surface of most differentiated cell types called primary cilium. Primary cilia are both chemo- and mechano-sensory in function and have an obvious role in cell cycle control. Because of this, it has been thought that primary cilia are not found on rapidly proliferating cells, for example, cancer cells. Here we report using immunofluorescent staining for the ciliary protein Arl13b that primary cilia are frequently found on HeLa (human epithelial adenocarcinoma) and other cancer cell lines such as MG63 (human osteosarcoma) commonly used for cell culture studies and that the ciliated population is significantly higher (ave. 28.6% and 46.5%, respectively in starved and 15.7-18.6% in un-starved cells) than previously anticipated. Our finding impacts the current perception of primary cilia formed in highly proliferative cells.
MeSH term(s)	ADP-Ribosylation Factors/metabolism ; Animals ; Cell Differentiation/physiology ; Cell Line, Tumor ; Cell Proliferation/physiology ; Cilia/metabolism ; Cilia/physiology ; Fluorescent Antibody Technique/methods ; HeLa Cells ; Humans ; Mice ; NIH 3T3 Cells ; Neoplasms/metabolism ; Neoplasms/pathology ; Neoplasms/physiopathology ; Signal Transduction
Chemical Substances	ADP-Ribosylation Factors (EC 3.6.5.2) ; ARL13B protein, human (EC 3.6.5.2)
Language	English
Publishing date	2015-08-06
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1143453-3
ISSN	1095-8355 ; 1065-6995
ISSN (online)	1095-8355
ISSN	1065-6995
DOI	10.1002/cbin.10500
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Article ; Online: Primary Cilia in Amacrine Cells in Retinal Development.

Ning, Ke / Sendayen, Brent E / Kowal, Tia J / Wang, Biao / Jones, Bryan W / Hu, Yang / Sun, Yang

Investigative ophthalmology & visual science

2021 Volume 62, Issue 9, Page(s) 15

Abstract: Purpose: Primary cilia are conserved organelles found in polarized cells within the eye that regulate cell growth, migration, and differentiation. Although the role of cilia in photoreceptors is well-studied, the formation of cilia in other retinal cell ...

Abstract	Purpose: Primary cilia are conserved organelles found in polarized cells within the eye that regulate cell growth, migration, and differentiation. Although the role of cilia in photoreceptors is well-studied, the formation of cilia in other retinal cell types has received little attention. In this study, we examined the ciliary profile focused on the inner nuclear layer of retinas in mice and rhesus macaque primates. Methods: Retinal sections or flatmounts from Arl13b-Cetn2 tg transgenic mice were immunostained for cell markers (Pax6, Sox9, Chx10, Calbindin, Calretinin, ChaT, GAD67, Prox1, TH, and vGluT3) and analyzed by confocal microscopy. Primate retinal sections were immunostained for ciliary and cell markers (Pax6 and Arl13b). Optical coherence tomography (OCT) and ERGs were used to assess visual function of Vift88 mice. Results: During different stages of mouse postnatal eye development, we found that cilia are present in Pax6-positive amacrine cells, which were also observed in primate retinas. The cilia of subtypes of amacrine cells in mice were shown by immunostaining and electron microscopy. We also removed primary cilia from vGluT3 amacrine cells in mouse and found no significant vision defects. In addition, cilia were present in the outer limiting membrane, suggesting that a population of Müller glial cells forms cilia. Conclusions: We report that several subpopulations of amacrine cells in inner nuclear layers of the retina form cilia during early retinal development in mice and primates.
MeSH term(s)	Amacrine Cells/ultrastructure ; Animals ; Chickens ; Cilia ; Electroretinography ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microscopy, Electron ; Models, Animal ; Rabbits ; Retina/growth & development ; Retina/ultrastructure ; Tomography, Optical Coherence/methods
Language	English
Publishing date	2021-07-21
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.62.9.15
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Article ; Online: OCRL regulates lysosome positioning and mTORC1 activity through SSX2IP-mediated microtubule anchoring.

Wang, Biao / He, Wei / Prosseda, Philipp P / Li, Liang / Kowal, Tia J / Alvarado, Jorge A / Wang, Qing / Hu, Yang / Sun, Yang

EMBO reports

2021 Volume 22, Issue 7, Page(s) e52173

Abstract: Lysosomal positioning and mTOR (mammalian target of rapamycin) signaling coordinate cellular responses to nutrient levels. Inadequate nutrient sensing can result in growth delays, a hallmark of Lowe syndrome. OCRL mutations cause Lowe syndrome, but the ... ...

Abstract	Lysosomal positioning and mTOR (mammalian target of rapamycin) signaling coordinate cellular responses to nutrient levels. Inadequate nutrient sensing can result in growth delays, a hallmark of Lowe syndrome. OCRL mutations cause Lowe syndrome, but the role of OCRL in nutrient sensing is unknown. Here, we show that OCRL is localized to the centrosome by its ASH domain and that it recruits microtubule-anchoring factor SSX2IP to the centrosome, which is important in the formation of the microtubule-organizing center. Deficiency of OCRL in human and mouse cells results in loss of microtubule-organizing centers and impaired microtubule-based lysosome movement, which in turn leads to mTORC1 inactivation and abnormal nutrient sensing. Centrosome-targeted PACT-SSX2IP can restore microtubule anchoring and mTOR activity. Importantly, boosting the activity of mTORC1 restores the nutrient sensing ability of Lowe patients' cells. Our findings highlight mTORC1 as a novel therapeutic target for Lowe syndrome.
MeSH term(s)	Animals ; Cell Cycle Proteins ; Humans ; Lysosomes ; Mechanistic Target of Rapamycin Complex 1/genetics ; Mice ; Microtubule-Associated Proteins ; Microtubules ; Oculocerebrorenal Syndrome ; Phosphoric Monoester Hydrolases
Chemical Substances	Cell Cycle Proteins ; Microtubule-Associated Proteins ; SSX2IP protein, human ; Ssx2ip protein, mouse ; Mechanistic Target of Rapamycin Complex 1 (EC 2.7.11.1) ; Phosphoric Monoester Hydrolases (EC 3.1.3.2) ; OCRL protein, human (EC 3.1.3.36) ; Ocrl protein, mouse (EC 3.1.3.36)
Language	English
Publishing date	2021-05-13
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2020896-0
ISSN	1469-3178 ; 1469-221X
ISSN (online)	1469-3178
ISSN	1469-221X
DOI	10.15252/embr.202052173
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Zs.A 5433: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Optogenetic stimulation of phosphoinositides reveals a critical role of primary cilia in eye pressure regulation.

Prosseda, Philipp P / Alvarado, Jorge A / Wang, Biao / Kowal, Tia J / Ning, Ke / Stamer, W Daniel / Hu, Yang / Sun, Yang

Science advances

2020 Volume 6, Issue 18, Page(s) eaay8699

Abstract: Glaucoma is a group of progressive optic neuropathies that cause irreversible vision loss. Although elevated intraocular pressure (IOP) is associated with the development and progression of glaucoma, the mechanisms for its regulation are not well ... ...

Abstract	Glaucoma is a group of progressive optic neuropathies that cause irreversible vision loss. Although elevated intraocular pressure (IOP) is associated with the development and progression of glaucoma, the mechanisms for its regulation are not well understood. Here, we have designed CIBN/CRY2-based optogenetic constructs to study phosphoinositide regulation within distinct subcellular compartments. We show that stimulation of CRY2-OCRL, an inositol 5-phosphatase, increases aqueous humor outflow and lowers IOP in vivo, which is caused by a calcium-dependent actin rearrangement of the trabecular meshwork cells. Phosphoinositide stimulation also rescues defective aqueous outflow and IOP in a Lowe syndrome mouse model but not in IFT88
Language	English
Publishing date	2020-04-29
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.aay8699
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