In order to explore the melanocortin receptor 1 gene (MC1R) in Amphilophus citrinellus embryo development and the formation of the body color expression location and function of the test preparation of MC1R gene RNA is antisense probes, T7 has sense direction of transcription of RNA probe concentration is 447.529 ng/μL, SP6 direction of transcription antisense RNA probe concentration is 342.698 ng/μL. After 10 to 20 times dilution probe was used for in situ hybridization, the MC1R gene probe expression positioning shows that A. citrinellus embryo developmental hybridization signal was weakening trend in general, the intestinal phase and bubble phase, yolk and lateral embryonic body parts have a hybridization signal distribution; in the film stage, hybridization signals appeared in the spine, yolk sac and yolk sac contents and pigment cells. There is a signal distribution on the yolk surface. In general, hybridization signals are specifically expressed in chordate, yolk sac and yolk sac contents and pigment cells, and the sense probe as a negative control group in five embryonic stages without any signal, sage MC1R expression positioning shows that the signal is revealed in A. citrinellus pigment cells that play an important regulating role in differentiation, migration, and are also associated with nerve, nutrition, immune function, preliminary establishment of fish body color related gene function and orientation of the entire embryo in situ hybridization method.