| Part1 Distribution of the calcium sensing receptor in eyes of normal rats1.Objective: To explore the expression of calcium sensing receptor(Ca SR) in normal rats' eyes.2.Methods: The eyes were obtained from the normal SD rats of 10 weeks old, and embedded in paraffin wax.The expression and distribution of Ca SR in eyeball were detected by immunohistochemistry. Quantitative RT-PCR and western blot were also used to probe for the presence of Ca SR in cornea,lens and retina of normal rats.3.Results: Ca SR immunoreactivity was detected in corneal endothelial cells and epithelial cells, lens epithelium, ciliary body and retina. Western blot and quantitative RT-PCR in rat ocular tissues has confirmed that Ca SR protein and m RNA were expressed in isolated cornea, retina, lens. Comparing the Ca SR protein and m RNA gene expression in eyes,cornea was more pronounced than lens,the retina was the fewest.Statistically significant difference between each part can be detected(t-test, p<0.05, Sigma Plot12.0).4.Conclusion: CaSR was first detected and widely expressed in rat ocular tissues.Part2 Decrease expression of Ca SR in diabetic eye disease1.Objective: To observe the expression of Ca SR in ocular tissues of diabetic rats.2.Methods: Twenty male Sprague Dawley(SD) rats were randomly divided into diabetic groups and control group. The diabetes mellitus models were successfully established by given intraperitoneal injection of STZ(60mg / Kg). Blood glucose was tested every four weeks.Twenty weeks later,six rats from both diabetic and control groups were randomly selected for q-RT-PCR and western blot. And the last three rats were used for Ca SR immunohistochemical staining.3.Results: Hyperglycemia was initially observed in the rats of diabetic group 3 days after administration. And diabetic rats had significantly lower body weights 20 weeks after STZ injection compared to control group.Additionally,we detected that the alteration of Ca SR was downregulated in ocular tissues through immunohistochemistry and westen blot analysis.Ca SR m RNA was also reduced in the diabetic rats' eyes by RT-PCR.4.Conclusion: The expression of Ca SR was widly decrease in ocular tissue of diabetic rats,which indicates that the expression of Ca SR was associated with diabetic eye disease. |
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