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The Journal of Neuroscience, October 1, 1998, 18(19):7625-7637
Cloning and Expression of Two Related Connexins from the Perch
Retina Define a Distinct Subgroup of the Connexin Family
John
O'Brien1,
Roberto
Bruzzone2,
Thomas W.
White3,
Muayyad R.
Al-Ubaidi1, and
Harris
Ripps1
1 Lions of Illinois Eye Research Institute, Department
of Ophthalmology and Visual Sciences, University of Illinois College of
Medicine, Chicago, Illinois 60612, 2 Unité de
Neurovirologie et Régénération du Système
Nerveux, Institut Pasteur, Paris, CEDEX 15, France, and
3 Department of Cell Biology, Harvard Medical School,
Boston, Massachusetts 02115
We have cloned cDNAs for two closely related connexins (Cx), Cx35
and Cx34.7, from a perch retinal cDNA library. Sequencing of PCR
products from genomic DNA revealed that both connexins have an intron
71 bp after the translation initiation site; in Cx35, the intron is 900 bp in length, whereas in Cx34.7 it is ~20 kb. Southern blots of
genomic DNA suggest that the two connexins represent independent single
copy genes. In Northern blots, Cx35 and Cx34.7 transcripts were
detected in retina and brain; Cx34.7 also showed a weak signal in
smooth muscle (gut) RNA. Antibodies against Cx35 labeled a 30 kDa band
on a Western blot of retinal membranes, and in histological sections,
the pattern of antibody recognition was consistent with labeling of
bipolar cells and unidentified processes in the inner plexiform and
nerve fiber layers. When expressed in Xenopus oocytes,
Cx35 and Cx34.7 formed homotypic gap junctions, but the junctional
conductance between paired oocytes expressing Cx35 was 10-fold greater
than that recorded for gap junctional channels formed by Cx34.7. The
homotypic gap-junctional channels were closed in a voltage-dependent
manner but with relatively weak voltage sensitivity. Heterotypic gap
junctions formed by Cx35 and Cx34.7 displayed junctional conductances
similar to those of Cx34.7 homotypic pairs and showed a slightly
asymmetric current-voltage relationship; the side expressing Cx35
exhibited a higher sensitivity to transjunctional potentials. An
analysis of the sequence and gene structure of the connexin family
revealed that perch Cx35 and Cx34.7, skate Cx35, and mouse Cx36
constitute a novel  subgroup.
Key words:
retinal connexins; cloning; connexin subgroup; oocytes; neurons; gap junctions; channels; intercellular
communication
Copyright © 1998 Society for Neuroscience 0270-6474/98/18197625-13$05.00/0
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