Cloning of prodynorphin cDNAs from the brain of Australian and African lungfish: implications for the evolution of the prodynorphin gene.
In mammals the opioids Met-enkephalin and Leu-enkephalin are derived from a common precursor, proenkephalin, and as a result these neuropeptides are co-localized in enkephalinergic neurons. The mammalian scheme for enkephalinergic networks is not universal for all classes of sarcopterygian vertebrates. In an earlier study, distinct Met- and Leu-enkephalin-positive neurons were detected in the central nervous system (CNS) of the African lungfish, Protopterus annectens. More recently, characterization of proenkephalin cDNAs separately cloned from the CNS of P. annectens and the Australian lungfish, Neoceratodus forsteri, revealed that the proenkephalin gene in these species encodes only Met-enkephalin-related opioids. In the current study a full-length prodynorphin cDNA (accession No. AY 445637) was cloned and sequenced from the CNS of N. forsteri. In addition to encoding alpha-neoendorphin, dynorphin A and dynorphin B sequences unique to the lungfish, two Leu-enkephalin sequences, flanked by paired basic amino acid proteolytic cleavage sites, were detected in this precursor. The partial sequence of a P. annectens prodynorphin cDNA (accession No. AY445638) also encoded a Leu-enkephalin sequence and a novel YGGFF sequence. The presence of the Leu-enkephalin sequence in the lungfish prodynorphin precursors would explain the origin of the distinct Leu-enkephalin-positive neurons found in the African lungfish CNS. The realization that Met-enkephalin and Leu-enkephalin can be derived from distinct opioid-coding precursor genes calls into question the interpretation of comparative immunohistochemical studies that have mapped 'enkephalinergic' networks in non-mammalian vertebrates.
Dores
Robert M
RM
University of Denver, Department of Biological Sciences, Denver, CO 80210, USA. rdores@du.edu
Sollars
Cristina
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Lecaude
Stephanie
S
Lee
Jenny
J
Danielson
Phillip
P
Alrubaian
Jasem
J
Lihrman
Isabelle
I
Joss
Jean M P
JM
Vaudry
Hubert
H
D000349:Africa; D000595:Amino Acid Sequence; D000818:Animals; D001315:Australia; D001483:Base Sequence; D005075:Biological Evolution; D001921:Brain / Q000502:physiology*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004743:Enkephalin, Leucine / Q000235:genetics; D004744:Enkephalin, Methionine / Q000235:genetics; D004745:Enkephalins / Q000235:genetics*; D005399:Fishes / Q000235:genetics*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D004745:Enkephalins; D011498:Protein Precursors; D004744:Enkephalin, Methionine; D004743:Enkephalin, Leucine; C033660:preproenkephalin