Maintenance and care of the C57BL/6 and BALB-c mice used in this study were in accordance with the National Institutes of Health Guidelines for the Use of Laboratory Animals and the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. All animals were maintained on a 12-hour light/12-hour dark cycle, and tissues were collected at the end of a dark cycle at the appropriate age. We collected the eye, brain, heart, lungs, liver, spleen, kidney, skeletal muscle, testes, skin, cardiac veins and arteries, and uterus from 7-month-old mice to study the expression of Cfh in different tissues. Eyeballs (6 to 8) were collected from 1-, 3-, 5-, 10-, 15-, 30-, 180-, and 300-day-old mice to study the expression dynamics of Cfh in whole eye. To study the distribution of this gene expression in eye tissue and its expression during aging, six to eight eyeballs per age were obtained from 35-, 60-, 180-, 300-, 420-, 500-, and 600-day-old mice, and the distalmost 3-mm optic nerve cut from the scleral surface of the eyeballs (ON), posterior segment (PS), retina, iris-ciliary body, lens, and corneal tissues were collected after dissection. The proximal optic nerve with chiasm was also collected after the brain was dissected from 180- and 300-day-old mice. Mouse embryonic cDNA was obtained from Clontech (Mountain View, CA).
After transcardiac perfusion of 90-day-old albino (BALB-c) mice, eyeballs and liver tissues were fixed in 4% paraformaldehyde and embedded in OCT, and cryosections were prepared for immunolocalization studies of CFH protein.