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| EFFECT OF REVERSIBLE ATP DEPLETION ON TIGHT-JUNCTION INTEGRITY IN LLC-PK1 CELLS |
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| Author(s): CANFIELD PE, GEERDES AM, MOLITORIS BA |
| Source: AMERICAN JOURNAL OF PHYSIOLOGY Volume: 261 Issue: 6 Pages: F1038-F1045 Part: Part 2 Published: DEC 1991 |
| Times Cited: 110 References: 44 |
| Abstract: To further understand and investigate how ischemia affects the tight junction we have developed a 2-h model of rapidly reversible ATP depletion and cellular injury in confluent LLC-PK1 monolayers. ATP depletion was achieved utilizing substrate-free medium containing 0.1-mu-M antimycin A (AA). Cellular ATP levels dropped rapidly to < 5% of control values, but recovery of ATP and cell morphology was possible even after 2 h of exposure to AA. Ruthenium red, an electron-dense marker of tight-junction integrity, was excluded from the tight junctions of control monolayers but penetrated cellular tight junctions during ATP depletion in a duration-dependent manner. Electrical resistance across the monolayers remained unchanged in control monolayers but decreased linearly during ATP depletion to 59% of control values. Transmonolayer movement of [H-3]mannitol increased from a control level of 7 to 13.5% during ATP depletion. Recovery of tight-junction integrity was demonstrated by a slowing of [H-3]mannitol transfer from the basolateral to the apical medium. The transfer rate in control monolayers was 0.0126%/min. During the initial 120 min of cellular recovery from 2 h of ATP depletion, the transfer rate was 0.0789%/min, but this decreased to 0.0045%/min between 2 and 4 h of recovery. In summary, physiology, biochemical, and morphological evidence indicates that reversible ATP depletion results in rapid opening of cellular tight junctions. After ATP-repletion physiological studies indicate a recovery of tight-junction integrity. |
| Document Type: Article |
| Language: English |
Addresses:
1. VET AFFAIRS MED CTR, 1055 CLERMONT ST, DENVER, CO 80220 USA
2. UNIV COLORADO, SCH MED, DEPT MED, DENVER, CO 80262 USA
3. UNIV COLORADO, SCH MED, DEPT CELLULAR & STRUCT BIOL, DENVER, CO 80262 USA |
| Publisher: AMER PHYSIOLOGICAL SOC, 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 |
| Subject Category: Physiology |
| IDS Number: GX687 |
| ISSN: 0002-9513 |
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