Figure 2
Presence of opsins in mammalian spermatozoa.
(a) Quantitative real-time PCR analysis of opsin mRNAs in mouse testes and liver. Mouse eyes were used as a positive control. (b) Quantitative real–time PCR analysis of opsin mRNAs in mature human spermatozoa. (c) A representative Western-blot analysis of rhodopsin in human spermatozoa using the monoclonal antibodies RET-1 and 1D4. Each lane contained 15 μg protein in total. (d) A representative Western-blot analysis of rhodopsin in eyes from rhodopsin-knockout (KO) and wild-type (WT) mice. Rhodopsin was extracted with the same lysis buffer used for human spermatozoa (see Methods). Each lane contained 15 μg protein in total. (e) A representative Western-blot analysis of rhodopsin in human and mouse spermatozoa using the monoclonal antibody RET-1. The lanes of the wild-type (WT) and rhodopsin-knockout (KO) mouse spermatozoa contained 30 μg protein in total and the lane of human spermatozoa contained 15 μg protein in total. (f) A representative Western-blot analysis of melanopsin in mouse eye and spermatozoa and in human spermatozoa. The lane of the eye contained 5 μg protein in total and the lanes of mouse and human spermatozoa contained 15 μg protein each. (g) A representative Western-blot analysis of encephalopsin in mouse eye and spermatozoa and in human spermatozoa. The lane of the eye contained 5 μg protein in total and the lanes of mouse and human spermatozoa contained 15 μg protein each. (h) A representative Western-blot analysis of neuropsin in human and mouse spermatozoa. Each lane contained 15 μg protein in total.
