Deciphering the origin of Met-enkephalin and Leu-enkephalin in Lobe-finned fish: cloning of Australian lungfish proenkephalin

Sollars, Cristina; Danielson, Phillip B.; Joss, Jean; Dores, Robert M.

doi:10.1016/s0006-8993(00)02567-1

Cited by 21 publications

(15 citation statements)

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“…During the characterization of a proenkephalin cDNA cloned from extracts of Australian lungfish brain [12], a 3′RACE product was amplified with the degenerate opioid primer POMC 1024 (fig. 1) that encoded the partial sequence of dynorphin A (R 226 –Q 237 ), the complete sequence of a dynorphin-B-like peptide (Y 240 –V 252 ) and a stop codon (t 1036 a 1037 a 1038 ).…”

Section: Resultsmentioning

confidence: 99%

“…2 Alignment of prodynorphin and proenkephalin sequences. The amino acid sequences of human proenkephalin [2], Australian lungfish proenkephalin [12], Australian lungfish prodynorphin (this study), the partial sequence of African lungfish prodynorphin (this study), B. marinu s prodynorphin [11], F. rubripes prodynorphin (FuguGenscan_15833), and human prodynorphin [4] were aligned by inserting the minimum number of gaps following the rules for alignment outlined by Dores et al [34]. Opioid positions in proenkephalin were designated as sites ‘A’ through ‘G’.…”

Section: Resultsmentioning

confidence: 99%

“…A partial African lungfish prodynorphin cDNA was amplified from a brain cDNA library constructed in Lambda Zap (Stratagene, La Jolla, Calif., USA). As described in a study by Sollars et al [12], mRNA extracted from the pooled brains of African lungfish was reverse transcribed into cDNA using Moloney murine leukemia virus reverse transcriptase, and the resulting cDNAs were then inserted into the Uni-Zap XR vector which contains the T3 and T7 primer sites flanking the insertion site. As described by Sollars et al [12], the opioid primer POMC 1024 and the vector primer T7 were used to amplify an internal region of the African lungfish prodynorphin cDNA that corresponded to a 553 –a 942 (fig.…”

Section: Methodsmentioning

confidence: 99%

“…The PCR products amplified from the freshly extracted African lungfish brain tissue were subcloned as previously described [12]. The plasmid DNA was isolated following alkaline lysis and binding to glass filter binding plates (Millipore, Billerica, Mass., USA), as described by Azadan et al [22].…”

Section: Methodsmentioning

confidence: 99%

“…Support for the latter conclusion comes from the cloning of proenkephalin cDNAs from the CNS of the Australian lungfish [12]and the African lungfish [13]. These lobe-finned fish proenkephalin cDNAs encode five Met-enkephalin sequences, an 8-amino-acid extended form of Met-enkephalin (YGGFMRSL), and a 7-amino-acid extended form of Met-enkephalin (YGGFMGY).…”

Section: Introductionmentioning

confidence: 99%

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Cloning of Prodynorphin cDNAs from the Brain of Australian and African Lungfish: Implications for the Evolution of the Prodynorphin Gene

et al. 2004

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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 α-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.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cloning of Prodynorphin cDNAs from the Brain of Australian and African Lungfish: Implications for the Evolution of the Prodynorphin Gene

et al. 2004

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Analyzing the evolution of the opioid/orphanin gene family

Dores

Lecaude

Bauer

et al. 2002

Mass Spectrometry Reviews

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Advances in molecular biology have made it possible to rapidly obtain the amino acid sequence of neuropeptide precursors-either by cloning and sequencing the cDNA that encodes the precursor, or by reconstructing the arrangement of exons and introns in a neuropeptide-coding gene through genomic approaches. The databases generated from these molecular approaches have been used to design probes to identify the cells that express the gene, or to ascertain the rate of expression of the gene, and even to predict the post-translational modifications that can generate functional neuropeptides from a biologically inert precursor. Although the power of these approaches is substantial, it is appreciated that a gene sequence or an mRNA sequence reflects the potential products that may be assembled in a secretory cell. To understand the functional capabilities of the secretory cell, the molecular genetics approaches must be combined with procedures that actually characterize the end-products generated by the secretory cell. Recent advances in two-dimensional gel electrophoresis and mass spectrometry now make it possible to analyze neuropeptides from a relatively small amount of tissue. These procedures can reveal novel end-products, tissue-specific endoproteolytic cleavage events, and developmental shifts in post-translational processing schemes. A gene family that illustrates all of these processes and the advantages of combining genomics with proteomics is the opioid/orphanin gene family.

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Opioids and Opioid Receptors in Fishes ☆

Herrero-Turrión¹

2017

Reference Module in Life Sciences

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Deciphering the origin of Met-enkephalin and Leu-enkephalin in Lobe-finned fish: cloning of Australian lungfish proenkephalin

Cited by 21 publications

References 21 publications

Cloning of Prodynorphin cDNAs from the Brain of Australian and African Lungfish: Implications for the Evolution of the Prodynorphin Gene

Cloning of Prodynorphin cDNAs from the Brain of Australian and African Lungfish: Implications for the Evolution of the Prodynorphin Gene

Analyzing the evolution of the opioid/orphanin gene family

Opioids and Opioid Receptors in Fishes ☆

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