They were then immediately perfused with a fixative of 2% paraformaldehyde and 2.5% glutaraldehyde in phosphate buffered saline, and the eyes were subsequently removed. In some cases, eyes were enucleated and fixed by immersion in the same fixative. The eyes were then bisected along the vertical meridian and embedded in an Epon- Araldite mixture, with sections cut at 1 mm thickness and stained with toluidine blue as previously described. ANR 94 Measurements of the outer nuclear layer thickness were taken as an index of retinal degeneration, and were obtained from 54 locations around the retina, as described elsewhere. Where the ONL was Arformoterol tartrate disrupted in focal regions, the ONL measurements were taken as close as possible to these disruptions. The number of cone nuclei at different ages was determined from the plastic sections by counting the cones in 10 microscopic fields of a 235-mm length of retina in a given section: 5 contiguous fields in the posterior retina superior to the optic nerve head and 5 fields in the inferior posterior retina as previously described. Liposomes were prepared from PalmitoylOleoylphosphatidylcholine and Cholesterol with a molar ratio of 3:2. Liposomes were treated with high vacuum overnight and rehydrated with 250 mM ammonium sulfate. HA-1077 was loaded into the liposomes by the remote loading method. After loading, ammonium sulfate was dialyzed against a solution of 10 mM Hepes and 140 mM NaCl. The concentration of total lipid was 50 mmoles/ml. HA-1077 concentration inside the liposomes was determined by breaking down a small volume of the liposomes with 90% isopropyl alcohol 75 mM HCl and measuring drug concentration by Nanodrop chromatography with UV absorbance at 320 nM, comparing to reference standards. By contrast, photopic ERG response amplitudes were significantly lower than normal at all ages, beginning by at least 6 weeks of age. These primarily cone-mediated responses were found with photopic and flicker ERG measurements. Thus, the cone-mediated retinal deficit was evident at about the same age as the behavioral motor dysfunction, but before any detectable changes in retinal histology. The early decrease in cone function led us to examine the retinas for evidence of loss of cones. By counting cone nuclei in the retinas of R6/2 and wild-type retinas, we found that the number was virtually the same in both genotypes at 6�C8 weeks of age. However, at 10�C11 weeks and 18�C19 weeks of age, the number of cones was reduced from normal by about 44% and 55%, respectively. This method of counting cones has been effective and accurate in many studies of mouse retinal degeneration. The time window between 6�C8 weeks of age represented a readily measured decline in photopic ERG responses at a time at which retinal cells were still intact.