Studies on endothelin release and Na,K transport in porcine lens

Mansim C. Okafor, Partha Mukhopadhyay, Nicholas A. Delamere

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


PURPOSE. In an earlier study it was reported that thrombin significantly reduces the rate of Na,K-adenosine triphosphatase (ATPase)-mediated ion transport by porcine lens. Because thrombin stimulates the release of endogenous endothelin (ET)-1 stores from some tissues, and because ET-1 can cause Na,K-ATPase inhibition, this study was designed to determine whether thrombin causes release of ET-1 from the lens. METHODS. Intact porcine lenses were incubated in Krebs solution. The concentration of ET-1 in the solution was determined by ELISA. The rate of Na,K-ATPase-dependent ion transport was determined by measurement of ouabain-sensitive 86Rb uptake. RESULTS. Thrombin (1 U/mL) reduced the rate of ouabain-sensitive 86Rb uptake by approximately 40%. PD145065 (2 μM), an ET receptor antagonist, abolished the inhibitory effect of thrombin on 86Rb uptake. Added alone, PD145065 did not alter 86Rb uptake. After an incubation period of 30 minutes, thrombin increased the concentration of ET-1 in the bathing medium in a dose-dependent manner. The time course of ET-1 appearance in the bathing medium of thrombin-treated lenses showed a peak at 30 minutes followed by a gradual decline. Consistent with the idea that release of ET-1 from the lens is tightly regulated, neither the calcium ionophore A23187 (1 μM) nor depolarization by potassium-rich solution caused significant release. However, exposing the lens to insulin (150 nM) significantly increased the appearance of ET-1 in the bathing medium. In parallel studies, mRNA for prepro-ET-1 was detected in the epithelium of freshly isolated lenses. CONCLUSIONS. The results of the study suggest that ET-1 is produced in porcine lens cells and that thrombin and insulin are capable of stimulating the release of ET-1 from the lens. Thrombin-induced inhibition of Na,K-ATPase-dependent ion transport may be mediated in part through the activation of ET-1 receptors by ET-1 released from the lens.

Original languageEnglish (US)
Pages (from-to)790-796
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Issue number3
StatePublished - 2002

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience


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