Abstract A mouse monoclonal antibody directed against chicken Gln 8-luteinizing hormone-releasing hormone (cLHRH-I) was developed and characterized. This antibody was used for the development of a competitive microtiter plate enzyme-linked immunosorbent assay for avian LHRH. The assay was validated for use with tissue and was used at a working range between 5 pg and 10 ng per sample. Using this procedure, cLHRH-I and II were assayed in whole brain extracts of Japanese Quail embryos. Samples were taken at regular intervals between Day 6 of incubation through Day 1 posthatch. There were 10 samples taken at each age with 2 replicates of the entire sampling regime. Data from males and females were pooled. LHRH concentrations were low, then rose to higher levels (15 pg/mg tissue) between Days 10 through 13 and decreased thereafter. These changes are likely to be correlated with the activation of the hypothalamic-pituitary-gonadal axis. This is particularly apparent in later embryonic development.