Mucin gene expression is not regulated by estrogen and/or progesterone in the ocular surface epithelia of mice.
OBJECTIVE
Dry eye syndrome is prevalent in post-menopausal women, and post-menopausal women secrete less mucus in their reproductive tracts. Using a mouse model, the purpose of this study was to determine if estrogen and/or progesterone regulates Muc4 and Muc5AC gene expression in the ocular surface epithelia, as the hormones do in reproductive tract epithelia.


METHODS
Adult C57BL/6 mice were ovariectomized, and 19 days later, pellets containing estrogen, progesterone, or a combination were inserted subcutaneously. Ocular surface and reproductive tract tissues were harvested following seven days of hormone treatment. A control group consisted of ovariectomized mice that received no hormone treatment. Real-time reverse transcription-polymerase chain reaction was used to determine the tissue expression levels of mucin mRNA of each treatment group relative to the control. Muc4 mRNA expression levels were determined for the reproductive tract, and both Muc4 and Muc5AC expression levels were determined for the ocular surface epithelia. Muc4 and Muc5AC gene expression in ocular surface and Muc4 in reproductive tract epithelia was demonstrated by In Situ hybridization, and Muc4 and Muc5AC protein was demonstrated in the epithelia of animals in the experimental groups.


RESULTS
The mRNA expression levels of Muc4 and Muc5AC and the immunofluorescence localization pattern in the ocular surface epithelia were not significantly different in any hormone treatment group when compared to the control ovariectomized group. By comparison, mice that were administered estrogen had a significant increase of Muc4 mRNA in the reproductive tract epithelia, progesterone given in combination with estrogen antagonized the upregulatory effects of estrogen in the reproductive tract, and the amount of Muc4 mRNA in the reproductive tract of progesterone-treated animals was not different from ovariectomized controls. Immunofluorescence localization of Muc4 in the reproductive tract epithelia of the experimental groups correlated to message levels, with lack of Muc4 protein detected in the control and progesterone groups.


CONCLUSIONS
In comparison to reproductive tract epithelia, Muc4 and Muc5AC are not hormonally regulated by estrogen or progesterone in the ocular surface epithelia of mice. These data demonstrate that regulation of epithelial mucin genes is tissue specific.
Lange
Christine
C
Schepens Eye Research Institute and Department of Ophthalmology, Harvard Medical School, 20 Staniford Street, Boston, MA 02114, USA.
Fernandez
Jolene
J
Shim
David
D
Spurr-Michaud
Sandra
S
Tisdale
Ann
A
Gipson
Ilene K
IK
D000818:Animals; D000951:Antigens, Neoplasm; D003228:Conjunctiva / Q000378:metabolism* / Q000473:pathology; D015352:Dry Eye Syndromes / Q000378:metabolism* / Q000473:pathology; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D051379:Mice; D008810:Mice, Inbred C57BL; D055271:Mucin 5AC; D018396:Mucin-1; D055264:Mucin-4; D009077:Mucins / Q000032:analysis / Q000235:genetics*; D010052:Ovariectomy; D011374:Progesterone / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis
D000951:Antigens, Neoplasm; D004967:Estrogens; C060192:MUC1 protein, human; C524658:MUC4 protein, human; C524659:Muc4 protein, mouse; C524679:Muc5ac protein, mouse; D055271:Mucin 5AC; D018396:Mucin-1; D055264:Mucin-4; D009077:Mucins; D012333:RNA, Messenger; D011374:Progesterone