Isolation and characterization of galectins in the mammalian retina.
OBJECTIVE
Previous studies have suggested that galectins may be involved in retinal adhesion and photoreceptor cell survival. To elucidate the underlying mechanisms, the authors isolated retinal galectins, determined their types and distributions, and investigated the validity of the hypothesis, using rat models.


METHODS
An antibody was prepared against a bovine retinal lectin that was isolated by use of a lactose-agarose column. cDNA of the lectin was isolated by screening of a bovine retinal cDNA library, using the antibody, and then was sequenced. The cDNAs of rat retinal galectins were also isolated by means of polymerase chain reaction and used to produce an antibody against recombinant galectin-3. Using the described antibodies, the authors examined the distributions of galectins in bovine and rat retinas, morphologic changes of rat retinas induced by the antibodies, and distributional changes of galectins in constant-light-exposed rat retinas.


RESULTS
The cDNAs of bovine galectin-1, rat galectin-1, and rat galectin-3 were isolated. Galectin-1 was found in various regions, including the retinal pigment epithelium, outer limiting membrane, and outer plexiform layer in bovine and rat retinas. Galectin-3 was increasingly detected in the cytoplasm of Müller cells after constant light exposure after an increase in its transcript. Retinal detachment and vacuolation of the outer plexiform layer were induced in rat eyes by intravitreous injection of the anti-galectin-1 antibody.


CONCLUSIONS
Galectin-1 may be involved in adhesion of the photoreceptor and outer plexiform layers by interacting with glycoconjugates with beta-galactoside residues in the interphotoreceptor matrix and synaptic cleft matrix. Galectin-3 may increase in Müller cells of a degenerative rat retina, probably through endogenous anti-apoptosis.
Uehara
F
F
Department of Ophthalmology, Kagoshima University Faculty of Medicine, Japan. fuehara@med5.kufm.kagoshima-u.ac.jp
Ohba
N
N
Ozawa
M
M
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D000943:Antigens, Differentiation / Q000302:isolation & purification* / Q000502:physiology; D015153:Blotting, Western; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D002846:Chromatography, Affinity; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000032:analysis; D005136:Eye Proteins / Q000302:isolation & purification* / Q000502:physiology; D037483:Galectin 1; D037502:Galectin 3; D006388:Hemagglutinins / Q000302:isolation & purification* / Q000502:physiology; D007124:Immunoenzyme Techniques; D007267:Injections; D008297:Male; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D012160:Retina / Q000737:chemistry* / Q000378:metabolism; D012163:Retinal Detachment / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014822:Vitreous Body
D000906:Antibodies; D000943:Antigens, Differentiation; D017931:DNA Primers; D018076:DNA, Complementary; D005136:Eye Proteins; D037483:Galectin 1; D037502:Galectin 3; D006388:Hemagglutinins