Abstract
Abstract: :
Purpose: To examine the expression of Kruppel–like factor 4 (KLF4) during development in mouse eyes. KLF4 is a transcription factor that contains three zinc fingers and like Sp1, interacts with GC boxes in the promoter elements of various genes. Methods: C57BL6 mice were set up for timed mating. Embryos on embryonic (E) days 10.5, 12.5, 15.5, and 18.5 and eyes from mice at birth, and on postnatal (P) days 7, 11, 15, 30, and 60 were collected, fixed in 4% paraformaldehyde and processed for paraffin sections. Immunohistochemistry was performed using a polyclonal antibody for KLF4. Digoxigenin–labeled DNA probes were prepared and in situ hybridization experiments were carried out to detect transcript of KLF4. RT–PCR was performed on RNA samples collected from the cornea, conjunctiva, and lens of adult mice (P60). Results: Moderate to strong nuclear staining of KLF4 was observed in the corneal epithelium, keratocytes, conjunctival epithelium, and subconjunctival fibroblasts from P11 to P60. The KLF4 expression in these cells was very low from E10.5 to P7. The level of KLF4 in the lens was minimal throughout all stages. A similar pattern of transcript level was observed by in situ hybridization. RT–PCR confirmed that KLF4 transcript was detected in the cornea and conjunctiva, but not in the lens. Conclusions: The expression of KLF4 in the mouse cornea and conjunctiva is spatially and temporally regulated. This transcription factor may be important for the development of these tissues.
Keywords: transcription factors • cornea: basic science • gene/expression