EGF receptor mRNA and protein in rat lacrimal acinar cells: evidence of its EGF-dependent phosphotyrosilation.
This study was designed to demonstrate the presence of the epidermal growth factor (EGF) receptor in the rat exorbital lacrimal gland. EGF receptor gene transcription was demonstrated 1) by reverse transcription-polymerase chain reaction analysis of lacrimal gland and acinar cells from RNA with a set of specific primers deduced from the rat EGF receptor sequence and 2) by Northern blot analysis of rat lacrimal gland mRNA. Lacrimal acinar cell preparations contain a low but detectable amount of specific 125I-EGF binding sites and efficiently internalize the ligand on binding at 37 degrees C. A sheep polyclonal antibody, directed against the human EGF receptor, detects a protein of 170 kDa by Western blot analysis of membrane proteins of the whole gland. This protein can be immunoprecipitated by the same antibody from whole gland membrane proteins as well as from solubilized acinar cells. Incubations of acinar cells in the presence of EGF results in an increased content of tyrosine-phosphorylated residues in immunoprecipitated 170-kDa protein. Taken together, these results demonstrate for the first time both EGF receptor gene transcription and protein expression in a lacrimal tissue, i.e., the rat exorbital lacrimal gland. These results also suggest a specific cellular location of the EGF receptor in a cell population contained in acinar cell preparations.
Marechal
H
H
Laboratoire de Biochimie des Transports Cellulaires, Centre National de la Recherche Scientifique, Unité de Recherche Associée 116, Université Paris-Sud, Orsay, France.
Jammes
H
H
Rossignol
B
B
Mauduit
P
P
D000818:Animals; D016375:Antisense Elements (Genetics) / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D004815:Epidermal Growth Factor / Q000502:physiology*; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D007765:Lacrimal Apparatus / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D019000:Phosphotyrosine / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic
D016375:Antisense Elements (Genetics); D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D019000:Phosphotyrosine; D004815:Epidermal Growth Factor; D066246:ErbB Receptors