Expression of amphiregulin in the sheep mammary gland.
The reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify, from sheep mammary gland total RNA, a 280 bp sequence of amphiregulin cDNA. Cloned and sequenced, it corresponded to the 78 amino acids of the major secreted form of amphiregulin, showing 81, 70 and 69% identity with human, rat and mouse amphiregulin, respectively. Expression of amphiregulin was detected by RT-PCR in the mammary gland at several developmental stages (fetal, lamb, early and late pregnant and lactating ewes) and in isolated myoepithelial cells. By Western blotting with an antiserum to human amphiregulin, two molecular weight forms, 27 and 51 kDa were detected in sheep mammary gland microsomal preparations, in a mammary gland extract after heparin affinity chromatography and in a medium conditioned by mammary epithelial cells. By immunocytochemistry, amphiregulin was detected in the cytoplasm and nuclei of luminal epithelial cells, myoepithelial cells and in intralobular stroma. An autocrine/paracrine role in sheep mammary growth is indicated.
Forsyth
I A
IA
Department of Cellular Physiology, Babraham Institute, Cambridge, UK.
Taylor
J A
JA
Keable
S
S
Turvey
A
A
Lennard
S
S
D000595:Amino Acid Sequence; D066258:Amphiregulin; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D066255:EGF Family of Proteins; D005260:Female; D015870:Gene Expression; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D006133:Growth Substances / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012194:RNA-Directed DNA Polymerase; D017422:Sequence Analysis, DNA; D012756:Sheep
C587848:AREG protein, human; D066258:Amphiregulin; C587849:Areg protein, mouse; C587850:Areg protein, rat; D018076:DNA, Complementary; D066255:EGF Family of Proteins; D006023:Glycoproteins; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D012194:RNA-Directed DNA Polymerase