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Vol. 11, Issue 2, 627-634, February 2000
and
Departments of *Cell Biology and Occludin and claudin are the major integral membrane components of
the mammalian tight junction. Although more than 11 distinct claudins
have been identified, only 1 occludin transcript has been reported thus
far. Therefore, we searched by reverse transcription-PCR for
occludin-related sequences in Madin-Darby canine kidney (MDCK) mRNA
and identified a transcript encoding an alternatively spliced form of
occludin, designated occludin 1B. The occludin 1B transcript contained
a 193-base pair insertion encoding a longer form of occludin with a
unique N-terminal sequence of 56 amino acids. Analysis of the MDCK
occludin gene revealed an exon containing the 193-base pair sequence
between the exons encoding the original N terminus and the distal
sequence, suggesting that occludin and occludin 1B arise from
alternative splicing of one transcript. To assess the expression and
distribution of occludin 1B, an antibody was raised against its unique
N-terminal domain. Immunolabeling of occludin 1B in MDCK cells revealed
a distribution indistinguishable from that of occludin. Furthermore,
occludin 1B staining at cell-to-cell contacts was also found in
cultured T84 human colon carcinoma cells and in frozen sections
of mouse intestine. Immunoblots of various mouse
tissues revealed broad coexpression of occludin 1B with occludin. The
wide epithelial distribution and the conservation across species
suggests a potentially important role for occludin 1B in the structure
and function of the tight junction.
Neurobiology,
Harvard Medical School, Boston, Massachusetts 02115
Corresponding author. E-mail address:
daniel_goodenough{at}hms.harvard.edu.
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