Expression cloning of gastric mucin complementary DNA and localization of mucin gene expression.
OBJECTIVE
Secretory mucins play an important role in gastric cytoprotection and are derived from a heterogeneous family of genes. The aim of this study was to determine the specific type and location of mucin gene expression in the human stomach.


METHODS
Expression cloning was performed by screening a human gastric complementary DNA expression library with antisera against deglycosylated gastric mucin. RNA analysis and immunohistochemistry were used to quantify and localize mucin gene expression.


RESULTS
Sequencing of positive clones revealed two clones containing tandem repeats. The first contained a 169-amino acid repeat and was named MUC6 (as previously described). The second contained the same 8-amino acid repeat consensus sequence (APTTSTTS) as complementary DNAs previously isolated from a tracheobronchial complementary DNA library and was labeled MUC5 (or MUC5AC). RNA analysis indicated that the gastric epithelium contains high levels of MUC5 and MUC6 messenger RNA with little or no MUC2, MUC3, and MUC4 messenger RNA. Immunohistochemical analysis showed that surface mucous cells of the cardia, fundus, and antrum expressed MUC5 peptide. In contrast, MUC6 peptide expression was limited to mucous neck cells of the fundus, antral-type glands of the antrum and cardia, and Brunner's glands of the duodenum.


CONCLUSIONS
MUC5 and MUC6 represent major secretory mucins in the stomach and are localized to distinct cell types.
Ho
S B
SB
Department of Medicine, VA Medical Center, Minneapolis, Minnesota, USA.
Roberton
A M
AM
Shekels
L L
LL
Lyftogt
C T
CT
Niehans
G A
GA
Toribara
N W
NW
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism*; D015870:Gene Expression; D015698:Genomic Library; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D055271:Mucin 5AC; D055257:Mucin-5B; D009077:Mucins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid
D018076:DNA, Complementary; C524678:MUC5AC protein, human; C524578:MUC5B protein, human; D055271:Mucin 5AC; D055257:Mucin-5B; D009077:Mucins; D012333:RNA, Messenger