Maintenance and care of the C57BL/6 and BALBc mice used in this study were in accordance with the NIH 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–dark cycle, and tissues were collected at the end of a dark cycle at the appropriate age. We collected the retina, lens, iris-ciliary body (I-CB), PS (containing the RPE, choroid, and sclera), and other tissues, including brain, heart, lungs, liver, spleen, kidney, skeletal muscle, testes, skin, and uterus from 33-week-old C57BL/6 mice, to isolate RNA and measure expression of the
Mfrp gene. Eyes were collected from 1-, 3-, 5-, 10- and 30-day-old C57BL/6 mice to study the expression of this gene during postnatal development. To measure the expression of
Mfrp during aging, eyes were obtained from 1-, 2-, 6-, 10-, 12-, 14-, 17- and 20-month-old C57BL/6 mice, and I-CB, PS, lens, and retina were dissected for RNA isolation. Heads from embryonic day (E)18 and postnatal day (P)1 to P10 C57BL/6 mice and eyes from adult (90-day-old) BALBc mice (albino) were fixed and processed for cryosectioning, as described earlier.
12 Ten-micrometer-thick radial cryosections were used for immunohistochemical analysis.