Cloning of proopiomelanocortin from the brain of the African lungfish, Protopterus annectens, and the brain of the western spadefoot toad, Spea multiplicatus

Jenny Lee, Stephanie Lecaude, Phillip Danielson, Cristina Sollars, Jasem Alrubaian, Catherine R Propper, Isabelle Lihrmann, Hubert Vaudry, Robert M. Dores

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

A degenerate primer, specific for the opioid core sequence YGGFM, was used to clone and sequence proopiomelanocortin (POMC) cDNAs from the brain of the African lungfish, Protopterus annectens, and from the brain of the western spadefoot toad, Spea multiplicatus. In addition, the opioid-specific primer was used to clone and sequence a 3'RACE product corresponding to a portion of the open reading frame of S. multiplicatus proenkephalin. For both species, cDNA was made from a single brain and a degenerate opioid-specific primer provided a reliable probe for detecting opioid-related cDNAs. The African lungfish POMC cDNA was 1168 nucleotides in length, and contained regions that are similar to tetrapod POMCs and fish POMCs. The African lungfish POMC encodes a tetrapod-like γ-MSH sequence that is flanked by sets of paired basic amino acid proteolytic cleavage sites. The γ-MSH region in ray-finned fish POMCs either has degenerate cleavage sites or is totally absent in some species. However, the African lungfish γ-MSH sequence does contain a deletion which has not been observed in tetrapod γ-MSH sequences. The β-endorphin region of lungfish POMC has the di-amino acid sequence tryptophan-aspartic acid in the N-terminal region and an additional glutamic acid residue in the C-terminal region of β-endorphin - features found in fish β-endorphin, but not tetrapod β-endorphins. The western spadefoot toad POMC was 1186 nucleotides in length, and exhibited an organizational scheme typical for tetrapod POMCs. However, the toad POMC did lack a paired basic amino acid proteolytic cleavage site N-terminal to the β-MSH sequence. Thus, like rat POMC, it is doubtful that β-MSH is an end product in either the toad brain or intermediate pituitary. At the amino acid level, the toad POMC had 76% sequence identity with Xenopus laevis POMC and 68% sequence identity with Rana ribidunda POMC. The use of these POMC sequences to assess phylogenetic relationships within anuran amphibians will be discussed. With respect to the fragment of S. multiplicatus proenkephalin cDNA, two metenkephalin sequences and the metenkephalin-RF sequence were found encoded in this fragment. As seen for X. laevis and R. ridibunda proenkephalin, a leuenkephalin sequence was not detected in the C-terminal region of the S. multiplicatus proenkephalin. The absence of a leuenkephalin sequence may be a common feature of anuran amphibian proenkephalins.

Original languageEnglish (US)
Pages (from-to)43-54
Number of pages12
JournalNeuroendocrinology
Volume70
Issue number1
DOIs
StatePublished - 1999

Fingerprint

Pro-Opiomelanocortin
Anura
Organism Cloning
Brain
Melanocyte-Stimulating Hormones
Endorphins
Complementary DNA
Opioid Analgesics
Basic Amino Acids
Xenopus laevis
Amphibians
Fishes
Nucleotides
Clone Cells
Skates (Fish)
Ranidae
Aspartic Acid
Tryptophan

Keywords

  • Amphibians
  • Corticotropin
  • Fishes
  • Melanostimulating hormone
  • Opiate peptides
  • Proopiomelanocortin

ASJC Scopus subject areas

  • Endocrinology
  • Neuroscience(all)

Cite this

Cloning of proopiomelanocortin from the brain of the African lungfish, Protopterus annectens, and the brain of the western spadefoot toad, Spea multiplicatus. / Lee, Jenny; Lecaude, Stephanie; Danielson, Phillip; Sollars, Cristina; Alrubaian, Jasem; Propper, Catherine R; Lihrmann, Isabelle; Vaudry, Hubert; Dores, Robert M.

In: Neuroendocrinology, Vol. 70, No. 1, 1999, p. 43-54.

Research output: Contribution to journalArticle

Lee, Jenny ; Lecaude, Stephanie ; Danielson, Phillip ; Sollars, Cristina ; Alrubaian, Jasem ; Propper, Catherine R ; Lihrmann, Isabelle ; Vaudry, Hubert ; Dores, Robert M. / Cloning of proopiomelanocortin from the brain of the African lungfish, Protopterus annectens, and the brain of the western spadefoot toad, Spea multiplicatus. In: Neuroendocrinology. 1999 ; Vol. 70, No. 1. pp. 43-54.
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T1 - Cloning of proopiomelanocortin from the brain of the African lungfish, Protopterus annectens, and the brain of the western spadefoot toad, Spea multiplicatus

AU - Lee, Jenny

AU - Lecaude, Stephanie

AU - Danielson, Phillip

AU - Sollars, Cristina

AU - Alrubaian, Jasem

AU - Propper, Catherine R

AU - Lihrmann, Isabelle

AU - Vaudry, Hubert

AU - Dores, Robert M.

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