Gene expression of the neuropeptide-processing enzyme carboxypeptidase E in rat photoreceptor cells.
We characterized the spatial expression of mRNA for the enzyme carboxypeptidase E (CPE) in the Long-Evans rat retina. CPE is involved in the processing of neuroactive peptides to a mature form. A cDNA encoding the 3' terminus of CPE mRNA was cloned by polymerase chain reaction amplification of rat retina single-stranded cDNA. The sequence of this cDNA was identical to a rat genomic clone for CPE and nearly identical (130/132 nucleotides) to a cDNA for rat brain CPE. In addition, the cDNA hybridized to a single allele on Southern blots and to a 2.1-kb mRNA on northern blots of both rat brain and retina. These data support the conclusion of others that CPE is a single-copy gene in the rat. In cell fractionation experiments, the majority of CPE mRNA fractionated with rod opsin mRNA, suggesting that CPE is expressed predominantly in rod photoreceptors. The high abundance of CPE mRNA in photoreceptors was confirmed by in situ hybridization studies, although CPE was detected at lower levels in other retinal cell types as well. The presence of abundant levels of the mRNA of a neuropeptide-processing enzyme in photoreceptor cells suggests that photoreceptors may utilize neuropeptides for normal function.
Nickells
R W
RW
Department of Visual Sciences, Indiana University, Bloomington.
Schlamp
C L
CL
Newton
A C
AC
Williams
D S
DS
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000201:enzymology; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000096:biosynthesis*; D004247:DNA / Q000032:analysis / Q000378:metabolism; D017931:DNA Primers; D015870:Gene Expression; D017403:In Situ Hybridization; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008969:Molecular Sequence Data; D010786:Photoreceptor Cells / Q000201:enzymology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D015183:Restriction Mapping; D013154:Spleen / Q000201:enzymology
D017931:DNA Primers; D012333:RNA, Messenger; D012313:RNA; D004247:DNA; D002268:Carboxypeptidases; D043423:Carboxypeptidase H