Human chorionic gonadotropin stimulates spheroid attachment on fallopian tube epithelial cells through the mitogen-activated protein kinase pathway and down-regulation of olfactomedin-1
FERTILITY AND STERILITY
Authors: So, Kam-Hei; Kodithuwakku, Suranga P.; Kottawatta, Kottawattage S. A.; Li, Raymond H. W.; Chiu, Philip C. N.; Cheung, Annie N. Y.; Ng, Ernest H. Y.; Yeung, William S. B.; Lee, Kai-Fai
Abstract
Objective: To study the effect of human chorionic gonadotropin (hCG) on olfactomedin-1 (Olfm1) expression and spheroid attachment in human fallopian tube epithelial cells in vitro. Design: Experimental study. Setting: Reproductive biology laboratory. Patient(s): Healthy nonpregnant women. Intervention(s): No patient interventions. Main Outcome Measure(s): Luteinizing hormone/chorionic gonadotropin receptor (LHCGR) and Olfm1 expression in fallopian tube epithelium cell line (OE-E6/E7 cells). OE-E6/E7 cells treated with hCG, U0126 extracellular signal-regulated kinase (ERK) inhibitor, or XAV939 Wnt/beta-catenin inhibitor were analyzed by Western blotting, real-time polymerase chain reaction, and in vitro spheroid attachment assay. Result(s): Human chorionic gonadotropin increased spheroid attachment on OE-E6/E7 cells through down-regulation of Olfm1 and activation of Wnt and mitogen-activated protein kinase (MAPK) signaling pathways. U0126 down-regulated both MAPK and Wnt/beta-catenin signaling pathways and up-regulated Olfm1 expression. XAV939 down-regulated only the Wnt/beta-catenin signaling pathway but up-regulated Olfm1 expression. Conclusion(s): Human chorionic gonadotropin activated both ERK and Wnt/beta-catenin signaling pathways and enhanced spheroid attachment on fallopian tube epithelial cells through down-regulation of Olfm1 expression. (C) 2015 by American Society for Reproductive Medicine.
Olfactomedin 2: Expression in the Eye and Interaction with Other Olfactomedin Domain-Containing Proteins
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
Authors: Sultana, Afia; Nakaya, Naoki; Senatorov, Vladimir V.; Tomarev, Stanislav I.
Abstract
PURPOSE. Olfactomedin 2 (OLFM2) belongs to the family of olfactomedin domain-containing proteins. Genetic data suggest its association with glaucoma in Japanese patients. However, its functions are still elusive. In this study, the properties of mammalian OLFM2 were investigated. METHODS. Expression of the rat and mouse Olfm2 gene was studied by using real-time PCR and in situ hybridization. Substitutions were introduced into OLFM2 by mutagenesis in vitro. Intracellular localization of OLFM2 was studied by confocal microscopy after transient transfection in HEK293 cells. Interaction of OLFM2 with olfactomedin 1 (Olfm1), olfactomedin 3 (Olfm3), myocilin, and gliomedin was studied by using co-immunoprecipitation. RESULTS. Two major human OLFM2 mRNAs encode secreted proteins with a length of 454 and 478 amino acids. OLFM2 is more closely related to OLFM1 and -3 than to any other family members. Olfm2 showed the most dynamic expression pattern compared with Olfm1 and -3 during mouse eye development and was expressed preferentially in the developing retinal ganglion cell layer. Among three OLFM2 substitutions tested (T86M, R144Q, and L420S), only L420S completely blocked secretion of the protein. OLFM2 interacted with Olfm1 and -3, but not with myocilin and gliomedin. Co-transfection of the L420S mutant with wild-type Olfm1 and -3 significantly inhibited secretion of Olfm1 and -3. CONCLUSIONS. Highly conserved OLFM2 protein may play an important role in the course of retinal and eye development. Severe mutations in one of the closely related olfactomedin domain-containing proteins (Olfm1-3) may block the secretion and probably the activity of all three family members, leading to more pronounced diseases of the retina than the knockout of individual genes. (Invest Ophthalmol Vis Sci. 2011;52:2584-2592) DOI:10.1167/iovs.10-6356
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