Objective: To construct the subtractive differentially expressed genes cDNA library of retina in rabbits with chronic ocular hypertension then to clone and screen the genes that related with retina damage.
Methods: Rabbit chronic ocular hypertension model was Established by injecting 1% methylcellulose into anterior chamber once a week for 5 weeks. Total RNA was extracted from retina of experimental and control rabbit eyes. Differentially expressed cDNAs libraries between chronic ocular hypertension eyes and control eyes were constructed using suppression subtraction hybridization (SSH) and then the preliminary data was screened for gene expression. One of upregulation expressed sequence tags (ESTs) was labeled with biotin as a probe. Cellular location of the interested genes in retina was identified by in situ hybridization.
Results: The constructed subtractive cDNA library that related to retina damage of chronic ocular hypertension was established and sixteen effective sequences were obtained. Two known expressed sequence tags (similarity over 98%) were found by BLAST Analyze in NCBI. The EST(number F9) was 99% similar to Ras genes family. Through ISH cellular location it was found the gene highly expressed in retina ganglion cells and inner nuclear layer of experimental retina but not in controls.
Conclusions: There was abnormal gene expression in rabbit retina in response to chronic ocular hypertension. Our result suggests EST F9 may play an important role in retina damage under chronic ocular hypertension. The subtractive differentially expressed genes cDNA library could provide valuable information in the research of optical neuro damage and protection in glaucoma.