Actin Filament Alterations in Rat Hepatocytes Induced In Vivo and In Vitro by Microcystin-LR, a Hepatotoxin from the Blue-green Alga, Microcystis aeruginosa

S. B. Hooser, V. R. Beasley, L. L. Waite, M. S. Kuhlenschmidt, W. W. Carmichael, W. M. Haschek

Research output: Contribution to journalArticlepeer-review

Abstract

The morphologic effects of microcystin-LR (MCLR) were examined in vitro and in vivo to identify the specific cell type(s) affected and to characterize the actin filament changes occurring in hepatocytes. Male Sprague Dawley rats were used for all studies. For in vitro studies, hepatic cells were isolated by collagenase perfusion of liver, while parenchymal cells (hepatocytes) and nonparenchymal cells were prepared by pronase digestion and metrimazide gradient centrifugation. Cell suspensions and primary hepatocyte monolayer cultures were treated with MCLR at doses up to 10 μg/ml; cultured hepatocytes were also treated with phalloidin or cytochalasin B at a dose of 10 μg/ml; and rats were treated intraperitoneally with MCLR at 180 mg/kg. Cultured hepatocyte preparations and frozen liver sections were stained with rhodamine-labeled phalloidin for filamentous actin. In cell suspensions. MCLR did not affect nonparenchymal cells but caused rapid, progressive, blebbing of the plasma membrane in hepatocytes. In cultured hepatocytes, MCLR caused plasma membrane blebbing as well as marked reorganization of actin microfilaments. These alterations were dose and time dependent. Cultured hepatocytes treated with phalloidin or cytochalasin B also showed extensive plasma membrane blebbing and actin filament alterations; however, actin filament changes were morphologically distinct from those induced by MCLR. In vivo, MCLR-induced hepatocyte actin alterations occurred at the same time as, or slightly preceded, histologic changes that began 30 minutes after dosing. These studies suggest that early MCLR-induced morphologic changes occurring both in vivo and in vitro are due to alterations in hepatocyte actin filaments.

Original languageEnglish (US)
Pages (from-to)259-266
Number of pages8
JournalVeterinary pathology
Volume28
Issue number4
DOIs
StatePublished - Jul 1991

Keywords

  • Actin filament
  • blue-green algae
  • hepatocyte
  • hepatotoxin
  • microcystin-LR
  • phalloidin
  • rat

ASJC Scopus subject areas

  • General Veterinary

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