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Am J Physiol Cell Physiol 276: C734-C746, 1999;
0363-6143/99 $5.00
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Vol. 276, Issue 3, C734-C746, March 1999

EGF precursor mRNA and membrane-associated EGF precursor protein in rat exorbital lacrimal gland

Hervé Maréchal1, Hélene Jammes2, Bernard Rossignol1, and Philippe Mauduit1

1 Laboratoire de Biochimie des Transports Cellulaires, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5619, Université Paris-Sud, 91405 Orsay Cedex; and 2 Unité d'Endocrinologie Moléculaire, Institut National de la Recherche Agronomique, 78352 Jouy en Josas Cedex, France

This study was designed to demonstrate the presence of epidermal growth factor (EGF) in the rat exorbital lacrimal gland. EGF precursor gene transcription was demonstrated first by RT-PCR analysis of lacrimal gland RNA using a set of specific primers and second by Northern blot analysis of rat lacrimal gland mRNA. A rabbit polyclonal antibody (rEGF2) directed against rat submaxillary gland EGF was used to detect EGF-containing proteins by RIA. Results indicate that the rat lacrimal gland does not contain detectable soluble and mature EGF but that the EGF immunoreactivity is associated with the membrane-enriched fraction. Analysis of the detergent-solubilized membrane proteins by gel filtration shows that membrane-associated EGF immunoreactivity was present as a high-molecular-mass protein. Moreover, as shown by Western blot analysis, a specific anti-rat EGF precursor antibody (ppEGF1) can immunoprecipitate a 152-kDa EGF-containing protein. Taken together, these results demonstrate for the first time both EGF precursor gene transcription and EGF precursor protein expression in a lacrimal tissue, i.e., the rat exorbital lacrimal gland. The demonstration that EGF appears to be stored only as its full-length membrane precursor may provide important information to study the regulation of its secretory process.

reverse transcriptase-polymerase chain reaction; rat epidermal growth factor antibody; rat epidermal growth factor precursor antibody; immunoprecipitation; exocrine gland


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