engleski

The expression of proliferating cell nuclear antigen and retinoblastoma protein in transplanted fetal rat lacrimal gland

A model of ectopic organogenesis of the rat fetal lacrimal gland was developed to study lacrimal gland organogenesis. It was done by its transplantation under the renal capsule. In transplants, expression of the tumor supresor retinoblastoma gene (Rb) and proliferating cell nuclear antigen (PCNA) was assessed at the protein level. Eyes of 17-days and 20-days-old rat fetuses were isolated under a dissecting microscope. Lacrimal glands were found and transplanted under the renal capsule of an adult male. After 14 days, transplants were routinely prepared for immunohistochemistry. DAKO Animal Research Kit, (Peroxidase) was used for detection of monoclonal mouse anti-PCNA and monoclonal mouse anti-human Retinoblastoma Gene Product. In transplants, teratoma-like structures developed that contained lacrimal gland epithelial cells and ducts as well as epidermis. PCNA signal was detected in nuclei of excretory duct cells and in epidermal basal layer cells. Some transplant cells were found have to have the ability of proliferation preserved even after the 14-day period. PCNA signal was absent in well-differentiated epithelial cells of lacrimal gland. Intranuclear Rb protein expression was only detected in several scattered cells, indicating the proliferating compartment to be usually larger than the differentiating one in fetal tissues. Assessment of the PCNA and Rb gene expression could prove important for elucidation of pathological processes in lacrimal gland, such as tumours or dry eye syndrome.

Lacrimal apparatus-embryology; Organ culture; Retinoblastoma protein-analysis; Animal

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