Molecular cloning of bullfrog prolactin receptor cDNA

Changes in prolactin receptor mRNA level during metamorphosis

Itaru Hasunuma, Kazutoshi Yamamoto, Sakae Kikuyama

    Research output: Contribution to journalArticle

    14 Citations (Scopus)

    Abstract

    In amphibian larvae, prolactin (PRL) is known to possess growth-promoting and anti-metamorphic activities. For further understanding of the role of PRL in larvae, bullfrog PRL receptor (bfPRLR) cDNA was obtained from the tail fin of premetamorphic tadpoles by use of the reverse transcription-polymerase chain reaction (RT-PCR) coupled with 5- and 3- rapid amplification of cDNA ends (RACE). The predicted bfPRLR was composed of 617 amino acids, contained a single transmembrane domain, and showed 33-57% sequence homologies with known sequences of vertebrate PRLRs. When bfPRLR was transiently expressed, specific binding of 125I-labeled bullfrog PRL (bfPRL) was observed. By Northern blot analysis, a 3-kb transcript was detected in the tail fin. By RT-PCR bfPRLR mRNA expression was detected mainly in the brain, kidney, skin, and tail throughout prometamorphic and middle climactic periods. The results of an RNase protection assay revealed that the bfPRLR mRNA level in the tail fin increased around the onset of climax (stage XX) and was maintained at a relatively high value at least until mid-climax (stage XXII). It also revealed that bfPRLR mRNA level in the kidney of larvae gradually rose as metamorphosis progressed. The results support the view that PRL in larval period acts not only on the larval organs but also on the organs that are necessary for the adult life to maintain or develop their structures and functions.

    Original languageEnglish
    Pages (from-to)200-210
    Number of pages11
    JournalGeneral and Comparative Endocrinology
    Volume138
    Issue number3
    DOIs
    Publication statusPublished - 2004 Sep 15

    Fingerprint

    Prolactin Receptors
    Rana catesbeiana
    Molecular Cloning
    metamorphosis
    molecular cloning
    prolactin
    Complementary DNA
    tail
    Messenger RNA
    receptors
    Prolactin
    fins
    Larva
    Tail
    larvae
    reverse transcriptase polymerase chain reaction
    kidneys
    Reverse Transcription
    rapid amplification of cDNA ends
    tadpoles

    Keywords

    • Bullfrog
    • Metamorphosis
    • Prolactin
    • Prolactin receptor

    ASJC Scopus subject areas

    • Endocrinology

    Cite this

    Molecular cloning of bullfrog prolactin receptor cDNA : Changes in prolactin receptor mRNA level during metamorphosis. / Hasunuma, Itaru; Yamamoto, Kazutoshi; Kikuyama, Sakae.

    In: General and Comparative Endocrinology, Vol. 138, No. 3, 15.09.2004, p. 200-210.

    Research output: Contribution to journalArticle

    Hasunuma, Itaru ; Yamamoto, Kazutoshi ; Kikuyama, Sakae. / Molecular cloning of bullfrog prolactin receptor cDNA : Changes in prolactin receptor mRNA level during metamorphosis. In: General and Comparative Endocrinology. 2004 ; Vol. 138, No. 3. pp. 200-210.
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    abstract = "In amphibian larvae, prolactin (PRL) is known to possess growth-promoting and anti-metamorphic activities. For further understanding of the role of PRL in larvae, bullfrog PRL receptor (bfPRLR) cDNA was obtained from the tail fin of premetamorphic tadpoles by use of the reverse transcription-polymerase chain reaction (RT-PCR) coupled with 5′- and 3′- rapid amplification of cDNA ends (RACE). The predicted bfPRLR was composed of 617 amino acids, contained a single transmembrane domain, and showed 33-57{\%} sequence homologies with known sequences of vertebrate PRLRs. When bfPRLR was transiently expressed, specific binding of 125I-labeled bullfrog PRL (bfPRL) was observed. By Northern blot analysis, a 3-kb transcript was detected in the tail fin. By RT-PCR bfPRLR mRNA expression was detected mainly in the brain, kidney, skin, and tail throughout prometamorphic and middle climactic periods. The results of an RNase protection assay revealed that the bfPRLR mRNA level in the tail fin increased around the onset of climax (stage XX) and was maintained at a relatively high value at least until mid-climax (stage XXII). It also revealed that bfPRLR mRNA level in the kidney of larvae gradually rose as metamorphosis progressed. The results support the view that PRL in larval period acts not only on the larval organs but also on the organs that are necessary for the adult life to maintain or develop their structures and functions.",
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