Different expression patterns for ubiquitous calpains and Capn3 splice variants in monkey ocular tissues.
The purpose of the present investigation was to compare the expression of ubiquitous and tissue-specific calpains in ocular tissues from the Macaca fascicularis monkey. Calpain isoforms in retina and corneal epithelium from adult M. fascicularis monkeys were characterized by RT-PCR, cDNA cloning and sequencing. Calpain isoform activities in ocular tissues were investigated by fractionation on DEAE-HPLC, immunoblotting, and casein zymography. Capn3 splice variants in the ocular tissues from rat, rabbit and monkey were compared after RT-PCR. RT-PCR analysis revealed that numerous splice variants of Capn3 were expressed in the epithelium from monkey cornea. The variants contained deletions or insertions in or around the IS1, IS2, and NS regions. The cDNAs for Capn3 variants were highly conserved, yet the expression patterns of the Capn3 isoforms were widely different among the mammalian species. In contrast, the expression patterns of ubiquitous calpains in ocular tissues were conserved among the mammalian species, and similarities between monkey and human cDNAs for Capn1 (mu-calpain) and Capn2 (m-calpain) were 98 and 99%, respectively. These results suggested that differences in expression patterns of Capn3 variants might be related to the function of each variant in a particular tissue or species.
Nakajima
T
T
Research Laboratory, Senju Pharmaceutical Company, Ltd., Kobe, Japan.
Fukiage
C
C
Azuma
M
M
Ma
H
H
Shearer
T R
TR
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D002154:Calpain / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000737:chemistry; D019573:Epithelium, Corneal / Q000201:enzymology; D019143:Evolution, Molecular; D005123:Eye / Q000201:enzymology*; D005786:Gene Expression Regulation; D007527:Isoenzymes; D008251:Macaca; D008969:Molecular Sequence Data; D009124:Muscle Proteins; D010446:Peptide Fragments / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000201:enzymology; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D007527:Isoenzymes; D009124:Muscle Proteins; D010446:Peptide Fragments; D020033:Protein Isoforms; D002154:Calpain; C491373:Capn3 protein, mouse; C491372:Capn3 protein, rat; C484806:CAPN1 protein, human