Functional Assessment of Intestinal Tight Junction Barrier and Ion Permeability in Native Tissue by Ussing Chamber Technique

Hempstock, Wendy; Ishizuka, Noriko; Hayashi, Hisaysohi

doi:10.3791/62468

Cited by 10 publications

(11 citation statements)

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“…Ussing chamber methods are the same as those described previously (Hempstock et al, 2021). Cells were cultured to confluent on 12-mm-diameter Transwell polycarbonate filters with 0.4-μm pore size (#3401; Corning).…”

Section: Ussing Chamber Electrophysiological Studiesmentioning

confidence: 99%

Reconstitution of functional tight junctions with individual claudin subtypes in epithelial cells

Furuse

Nakatsu

Hempstock

et al. 2023

Cell Struct. Funct.

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The claudin family of membrane proteins is responsible for the backbone structure and function of tight junctions (TJs), which regulate the paracellular permeability of epithelia.It is thought that each claudin subtype has its own unique function and the combination of expressed subtypes determines the permeability property of each epithelium. However, many issues remain unsolved in regard to claudin functions, including the detailed functional differences between claudin subtypes and the effect of the combinations of specific claudin subtypes on the structure and function of TJs. To address these issues, it would be useful to have a way of reconstituting TJs containing only the claudin subtype(s) of interest in epithelial cells. In this study, we attempted to reconstitute TJs of individual claudin subtypes in TJ-deficient MDCK cells, designated as claudin quinKO cells, which were previously established from MDCK II cells by deleting the genes of claudin-1, -2, -3, -4, and -7. Exogenous expression of each of claudin-1, -2, -3, -4, and -7 in claudin quinKO cells resulted in the reconstitution of functional TJs. These TJs did not contain claudin-12 and -16, which are endogenously expressed in claudin quinKO cells. Furthermore, overexpression of neither claudin-12 nor claudin-16 resulted in the reconstitution of TJs, demonstrating the existence of claudin subtypes lacking TJ-forming activity in epithelial cells. Exogenous expressions of the channel-forming claudin-2, -10a, -10b, and -15 reconstituted TJs with reported paracellular channel properties, demonstrating that these claudin subtypes form paracellular channels by themselves without interaction with other subtypes. Thus, the reconstitution of TJs in claudin quinKO cells is advantageous for further investigations of claudin functions.

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Section: Ussing Chamber Electrophysiological Studiesmentioning

confidence: 99%

Reconstitution of functional tight junctions with individual claudin subtypes in epithelial cells

Furuse

Nakatsu

Hempstock

et al. 2023

Cell Struct. Funct.

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“…As Ussing Chamber is widely used to study ion, drug, and nutrient transport in intestinal epithelium ( He et al, 2013 ; Hempstock et al, 2021 ; Michiba et al, 2021 ), and we applied it to investigate the regulation of Gln transport. In our study the Gln-induced ileal I sc well represents the Na + /Gln co-transport since, 1) the mucosal application of Gln induced a significant I sc , but the serosal application could not induce any detectable I sc ; 2) lowering the concentration of Na + in mucosal solution significantly reduced the Gln-induced I sc ; 3) either cinromide, a selective inhibitor of epithelial NGcT (B 0 AT1), or GPNA, a non-specific inhibitor of NGcT, attenuated Gln-stimulated I sc peak after added to the mucosal side.…”

Section: Discussionmentioning

confidence: 99%

Ca2+-Permeable Channels/Ca2+ Signaling in the Regulation of Ileal Na+/Gln Co-Transport in Mice

et al. 2022

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Oral glutamine (Gln) has been widely used in gastrointestinal (GI) clinical practice, but it is unclear if Ca2+ regulates intestinal Gln transport, although both of them are essential nutrients for mammals. Chambers were used to determine Gln (25 mM)-induced Isc through Na+/Gln co-transporters in the small intestine in the absence or the presence of selective activators or blockers of ion channels and transporters. Luminal but not serosal application of Gln induced marked intestinal Isc, especially in the distal ileum. Lowering luminal Na+ almost abolished the Gln-induced ileal Isc, in which the calcium-sensitive receptor (CaSR) activation were not involved. Ca2+ removal from both luminal and serosal sides of the ileum significantly reduced Gln- Isc. Blocking either luminal Ca2+ entry via the voltage-gated calcium channels (VGCC) or endoplasmic reticulum (ER) release via inositol 1,4,5-triphosphate receptor (IP3R) and ryanodine receptor (RyR) attenuated the Gln-induced ileal Isc, Likewise, blocking serosal Ca2+ entry via the store-operated Ca2+ entry (SOCE), TRPV1/2 channels, and Na+/Ca2+ exchangers (NCX) attenuated the Gln-induced ileal Isc. In contrast, activating TRPV1/2 channels enhanced the Gln-induced ileal Isc. We concluded that Ca2+ signaling is critical for intestinal Gln transport, and multiple plasma membrane Ca2+-permeable channels and transporters play roles in this process. The Ca2+ regulation of ileal Na+/Gln transport expands our understanding of intestinal nutrient uptake and may be significant in GI health and disease.

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“…− [58]. The composition of this solution is not well-defined for corrosion testing of metallic implants [59], resulting in substantially different corrosion resistance.…”

Section: Mg Corrosion In Simulated Body Environmentsmentioning

confidence: 99%

Biodegradable Magnesium Biomaterials—Road to the Clinic

Amukarimi

Mozafari

2022

Bioengineering

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In recent decades, we have witnessed radical changes in the use of permanent biomaterials. The intrinsic ability of magnesium (Mg) and its alloys to degrade without releasing toxic degradation products has led to a vast range of applications in the biomedical field, including cardiovascular stents, musculoskeletal, and orthopedic applications. With the use of biodegradable Mg biomaterials, patients would not suffer second surgery and surgical pain anymore. Be that as it may, the main drawbacks of these biomaterials are the high corrosion rate and unexpected degradation in physiological environments. Since biodegradable Mg-based implants are expected to show controllable degradation and match the requirements of specific applications, various techniques, such as designing a magnesium alloy and modifying the surface characteristics, are employed to tailor the degradation rate. In this paper, some fundamentals and particular aspects of magnesium degradation in physiological environments are summarized, and approaches to control the degradation behavior of Mg-based biomaterials are presented.

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Functional Assessment of Intestinal Tight Junction Barrier and Ion Permeability in Native Tissue by Ussing Chamber Technique

Cited by 10 publications

References 0 publications

Reconstitution of functional tight junctions with individual claudin subtypes in epithelial cells

Reconstitution of functional tight junctions with individual claudin subtypes in epithelial cells

Ca2+-Permeable Channels/Ca2+ Signaling in the Regulation of Ileal Na+/Gln Co-Transport in Mice

Biodegradable Magnesium Biomaterials—Road to the Clinic

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