Molecular probing of the secretory pathway in peptide hormone-producing cells.
The biosynthetic machinery in the melanotrope cells of the Xenopus intermediate pituitary is primarily dedicated to the generation of proopiomelanocortin (POMC)-derived, melanophore-stimulating peptides. Transfer of the animal to a black background stimulates the production of these peptides and causes a dramatic increase in POMC mRNA levels. To identify genes involved in the biosynthesis and regulated release of peptide hormones, we differentially screened an intermediate pituitary cDNA library of toads adapted to a black background with cDNA probes derived from intermediate pituitary mRNA of black- and white-adapted animals. Here we report the identification of twelve distinct genes whose expression levels in the melanotropes are regulated in coordination with that of POMC. Four of these genes are novel while the others code for translocon-associated proteins, a lumenal cysteine protease of the endoplasmic reticulum, prohormone-processing enzymes, members of the granin family and a transmembrane protein presumably involved in the assembly and/or specific functioning of vacuolar H(+)-ATPase from secretory granules. Our results indicate that a wide variety of both soluble and membrane-associated components of the secretory pathway is recruited in physiologically activated, peptide hormone-producing cells.
Holthuis
J C
JC
Department of Animal Physiology, University of Nijmegen, The Netherlands.
Jansen
E J
EJ
van Riel
M C
MC
Martens
G J
GJ
D000595:Amino Acid Sequence; D000818:Animals; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D005786:Gene Expression Regulation; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D011333:Pro-Opiomelanocortin / Q000235:genetics / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D014981:Xenopus / Q000378:metabolism*
D018076:DNA, Complementary; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin; D006180:Proton-Translocating ATPases