Expression changes in mRNAs and mitochondrial damage in lens epithelial cells with selenite

P. B. Belusko, T. Nakajima, M. Azuma, Thomas (Tom) Shearer

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

An overdose of sodium selenite induces cataracts in young rats. The mid-stage events producing the cataract include calpain-induced hydrolysis and precipitation of lens proteins. Apoptosis in lens epithelial cells has been suggested as an initial event in selenite cataracts. Expression levels of two genes associated with apoptosis were altered in lens epithelial cells from selenite-injected rats. The purpose of the present experiment was to perform a more comprehensive search for changes in expression of mRNAs in lens epithelial cells in order to more fully delineate the early events in selenite-induced cataracts. Lens epithelial cells were harvested at 1 and 2 days after a single subcutaneous injection of sodium selenite (30 μmol/kg body weight) into 12-day-old rats. Gene expression was analyzed using a commercial DNA array (Rat Genome U34A GeneChip array, Affymetrix). Of approximately 8000 genes assayed by hybridization, 13 genes were decreased and 27 genes were increased in the rat lens epithelial cells after injection of selenite. Some of the up-regulated genes included apoptosis-related genes, and a majority of the down-regulated genes were mitochondrial genes. Previously observed changes in expression of EGR-1 mRNA were also confirmed. Changes in the expression patterns of mRNAs were also confirmed by RT-PCR. To determine the mechanism for damage of lens epithelial cells (alpha TN4 cell) by culture in selenite, leakage of cytochrome c from mitochondria was measured. Selenite caused significant leakage of cytochrome c into the cytosol of alpha TN4 cells. Our data suggested that the loss of integrity of lens epithelial cells by selenite might be caused by preferential down-regulation of mitochondrial RNAs, release of cytochrome c, and impaired mitochondrial function. Up-regulation of mRNAs involved in maintenance of DNA, regulation of metabolism, and induction of apoptosis may also play roles.

Original languageEnglish (US)
Pages (from-to)135-142
Number of pages8
JournalBiochimica et Biophysica Acta - General Subjects
Volume1623
Issue number2-3
DOIs
StatePublished - Oct 13 2003

Fingerprint

Selenious Acid
Lenses
Genes
Epithelial Cells
Cataract
Rats
Cytochromes c
Apoptosis
Sodium Selenite
Messenger RNA
Crystallins
Calpain
Mitochondrial Genes
Subcutaneous Injections
mitochondrial messenger RNA
Oligonucleotide Array Sequence Analysis
Mitochondria
Cytosol
DNA
Hydrolysis

Keywords

  • DNA microarray
  • Lens epithelium
  • Mitochondria
  • Selenite

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Expression changes in mRNAs and mitochondrial damage in lens epithelial cells with selenite. / Belusko, P. B.; Nakajima, T.; Azuma, M.; Shearer, Thomas (Tom).

In: Biochimica et Biophysica Acta - General Subjects, Vol. 1623, No. 2-3, 13.10.2003, p. 135-142.

Research output: Contribution to journalArticle

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