Molecular cloning of newt prolactin (PRL) cDNA: Effect of temperature on PRL mRNA expression

N. Takahashi, I. Hasunuma, T. Iwata, K. Umezawa, K. Yamamoto, A. Marin, I. Perroteau, C. Vellano, S. Kikuyama

    Research output: Contribution to journalArticle

    10 Citations (Scopus)

    Abstract

    A partial prolactin (PRL) cDNA was specifically PCR amplified from a cDNA library constructed from pituitary mRNAs of the newt (Cynops pyrrhogaster) and cloned into plasmid vectors. One clone thus obtained contained a 739-bp insert encoding the C-terminal amino acid sequence of the mature hormone molecule. Using this clone as a probe, the full-length newt PRL cDNA was screened from the cDNA library. The PRL cDNA clone thus obtained consisted of 1024 bp encoding the entire sequence of the mature PRL molecule in addition to its signal peptide. The amino acid sequence of newt PRL deduced from its nucleotide sequence showed higher homologies with those PRL sequences of tetrapod animals than with those of teleosts. Northern blot analysis revealed the newt PRL mRNA size to be approximately 1 kb. In situ hybridization using the newt PRL cDNA as a probe revealed that the pituitary region expressing PRL mRNA corresponded to that immunoreactive with antiserum against PRL. PRL mRNA levels in the pituitary of newts subjected to room and low temperatures were determined by Northern analysis employing the PRL cDNA as a probe. PRL mRNA levels were significantly higher in the pituitaries of newts subjected to 10° than in those of newts kept at 23°. Likewise, immunoassayable plasma PRL levels were higher in animals subjected to 10° than in those kept at 23°.

    Original languageEnglish
    Pages (from-to)188-195
    Number of pages8
    JournalGeneral and Comparative Endocrinology
    Volume121
    Issue number2
    DOIs
    Publication statusPublished - 2001

    Keywords

    • Cynops pyrrhogaster
    • Low temperature
    • Prolactin cDNA
    • Prolactin mRNA expression
    • Red-bellied newt

    ASJC Scopus subject areas

    • Endocrinology

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  • Cite this

    Takahashi, N., Hasunuma, I., Iwata, T., Umezawa, K., Yamamoto, K., Marin, A., Perroteau, I., Vellano, C., & Kikuyama, S. (2001). Molecular cloning of newt prolactin (PRL) cDNA: Effect of temperature on PRL mRNA expression. General and Comparative Endocrinology, 121(2), 188-195. https://doi.org/10.1006/gcen.2000.7594