Electrophysiologic Analysis of Tight Junction Size and Charge Selectivity
© 2021 Wiley Periodicals LLC..
Tight junctions form selectively permeable barriers that limit paracellular flux across epithelial-lined surfaces. Rather than being absolute barriers, tight junctions in many tissues allow ions, water, and other small molecules to cross on the basis of size and charge selectivity via the high-capacity pore pathway. Most probes currently used to assess tight junction permeability exceed the maximum size capacity of the pore pathway. As a result, available analytical tools have generally been limited to measurement of transepithelial electrical resistances. These provide no information regarding size selectivity and, therefore, cannot be used to distinguish between the pore pathway and the leak pathway, a low-capacity route that accommodates larger macromolecules. This article describes use of dilution potential and bi-ionic potential measurements for analysis of tight junction size and charge selectivity within monolayers of cultured epithelial cells. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Culture of MDCK monolayers on semipermeable supports and induction of claudin-2 expression Basic Protocol 2: Configuring voltage/current clamp and other equipment Basic Protocol 3: Measuring dilution and bi-ionic potentials Basic Protocol 4: Calculating ion permeabilities and pore diameter Support Protocol: Preparation of agar bridges and electrophysiology rig setup.
| Errataetall: | |
|---|---|
| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:1 |
|---|---|
| Enthalten in: |
Current protocols - 1(2021), 6 vom: 15. Juni, Seite e143 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Shashikanth, Nitesh [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Barrier function |
|---|
| Anmerkungen: |
Date Completed 16.06.2021 Date Revised 16.07.2022 published: Print ErratumIn: Curr Protoc. 2021 Sep;1(9):e259. - PMID 34495581 Citation Status MEDLINE |
|---|
| doi: |
10.1002/cpz1.143 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM326545859 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM326545859 | ||
| 003 | DE-627 | ||
| 005 | 20231225194829.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1002/cpz1.143 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1088.xml |
| 035 | |a (DE-627)NLM326545859 | ||
| 035 | |a (NLM)34106526 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Shashikanth, Nitesh |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Electrophysiologic Analysis of Tight Junction Size and Charge Selectivity |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 16.06.2021 | ||
| 500 | |a Date Revised 16.07.2022 | ||
| 500 | |a published: Print | ||
| 500 | |a ErratumIn: Curr Protoc. 2021 Sep;1(9):e259. - PMID 34495581 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2021 Wiley Periodicals LLC. | ||
| 520 | |a Tight junctions form selectively permeable barriers that limit paracellular flux across epithelial-lined surfaces. Rather than being absolute barriers, tight junctions in many tissues allow ions, water, and other small molecules to cross on the basis of size and charge selectivity via the high-capacity pore pathway. Most probes currently used to assess tight junction permeability exceed the maximum size capacity of the pore pathway. As a result, available analytical tools have generally been limited to measurement of transepithelial electrical resistances. These provide no information regarding size selectivity and, therefore, cannot be used to distinguish between the pore pathway and the leak pathway, a low-capacity route that accommodates larger macromolecules. This article describes use of dilution potential and bi-ionic potential measurements for analysis of tight junction size and charge selectivity within monolayers of cultured epithelial cells. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Culture of MDCK monolayers on semipermeable supports and induction of claudin-2 expression Basic Protocol 2: Configuring voltage/current clamp and other equipment Basic Protocol 3: Measuring dilution and bi-ionic potentials Basic Protocol 4: Calculating ion permeabilities and pore diameter Support Protocol: Preparation of agar bridges and electrophysiology rig setup | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a barrier function | |
| 650 | 4 | |a claudin | |
| 650 | 4 | |a ion conductance | |
| 650 | 4 | |a permeability | |
| 650 | 4 | |a tight junction | |
| 650 | 7 | |a Claudin-2 |2 NLM | |
| 700 | 1 | |a Rizzo, Heather E |e verfasserin |4 aut | |
| 700 | 1 | |a Pongkorpsakol, Pawin |e verfasserin |4 aut | |
| 700 | 1 | |a Heneghan, John F |e verfasserin |4 aut | |
| 700 | 1 | |a Turner, Jerrold R |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Current protocols |d 2021 |g 1(2021), 6 vom: 15. Juni, Seite e143 |w (DE-627)NLM320466582 |x 2691-1299 |7 nnns |
| 773 | 1 | 8 | |g volume:1 |g year:2021 |g number:6 |g day:15 |g month:06 |g pages:e143 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1002/cpz1.143 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 1 |j 2021 |e 6 |b 15 |c 06 |h e143 | ||