Characterization and localization of an aldehyde dehydrogenase to amacrine cells of bovine retina.
An enzyme of bovine retina that catalyzes oxidation of retinaldehyde to retinoic acid was purified to homogeneity and a monoclonal antibody (mAb H-4) was generated. MAb H-4 recognized a single component (Mr = 55,000) in extracts of bovine retina and other bovine tissues. The antibody showed no cross-reactivity with extracts of rat, monkey, or human retinas. A 2067 bp cDNA was selected from a retina cDNA expression library using mAb H-4. The cDNA hybridized with a similarly sized, moderately abundant mRNA prepared from bovine retina. Nucleotide sequence analysis indicated that the cDNA contained a single open reading frame encoding 501 amino acids that have 88% sequence identity with the amino-acid sequence of human hepatic Class 1 aldehyde dehydrogenase. Amino-acid sequence analysis of purified enzyme demonstrated that the cDNA encodes the isolated enzyme. MAb H-4 specifically labeled the somata and processes of a subset of amacrine cells in bovine retinal sections. Labeled amacrine somata were located on both sides of the inner plexiform layer, and their processes ramified into two laminae within the inner plexiform layer. The inner radial processes of Müller (glial) cells were weakly reactive with mAb H-4. Weak immunostaining of amacrine cells was found in monkey retina with mAb H-4, but no signal was detected in rat or human retina. The results provide further evidence for metabolism and function of retinoids within cells of the inner retina and define a novel class of retinal amacrine cells.
Saari
J C
JC
Department of Ophthalmology, University of Washington, Seattle 98195, USA.
Champer
R J
RJ
Asson-Batres
M A
MA
Garwin
G G
GG
Huang
J
J
Crabb
J W
JW
Milam
A H
AH
D000444:Aldehyde Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D003001:Cloning, Molecular; D003429:Cross Reactions; D018076:DNA, Complementary / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D008254:Macaca nemestrina; D008969:Molecular Sequence Data; D009474:Neurons / Q000201:enzymology*; D051381:Rats; D012160:Retina / Q000201:enzymology*; D012172:Retinaldehyde / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014212:Tretinoin / Q000378:metabolism
D018076:DNA, Complementary; D014212:Tretinoin; D000444:Aldehyde Dehydrogenase; D012172:Retinaldehyde