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Mutant C57BL/6 mouse with corneal opacity (B6-Co) appears eye open at birth (EOB) phenotype, which is a good animal model in the study of developmental mechanism of eyelid. Investigating the relationship between serum response factor (SRF) and EOB phenotype can provide theoretical support for the research on the mechanism of innate defects in eyelid development in humans.
This study was to assess the dynamic expressions of SRF in eyelid of embryonic B6-Co mouse.
Total RNA was extracted from B6 and B6-Co mice eyelid tissue at embryonic day 16. 5 (E16. 5 d), E17. 5 d and E18. 5 d. The relative expression levels of SRF mRNA and protein in the eyelid tissue of B6 and B6-Co embryonic mice were assayed by real-time quantitative PCR and Western blot, respectively. In situ expressions of SRF protein in eyelid of B6-Co mice and B6 mice were detected using immunofluorescence technique. The use and care of the animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals of Nantong University.
The relative expression levels of SRF mRNA in the eyelids were 0. 41±0. 06 and 0. 24±0. 17 in E16. 5 d and E17. 5 d of B6-Co mice, showing a significant decline in comparison with 1. 03±0. 17 and 1. 01±0. 09 in the B6 mice (P=0. 025, 0. 017). The expression levels of SRF protein in the eyelids of E16. 5 d and E17. 5 d B6-Co mice were 0. 08±0. 01 and 0. 08±0. 01, which were significantly lower than 0. 12±0. 03 and 0. 13±0. 02 of B6 mice (P=0. 036, 0. 024). However, there were no significant differences in the expression levels of SRF mRNA and protein in E18. 5 d between the B6-Co mice and B6 mice (P=0. 387, 0. 774). Immunofluorescence assay displayed that SRF was expressed in the keratinocytes of eyelids in both mice, but the fluorescence intensity was weaker in the B6-Co mice.
SRF probably interrupts the developing process of eyelid in early embryo of B6-Co mice.