Abstract: Aim of this study was determination of sperms’ ATP content of golden grey mullet in different time, temperature and extenders. Caspian Sea Mugilidea is one of the most important fish of the sea fishery which nowadays is the predominant catches fish of the mentioned sea. The ATP content of mentioned fish sperm is an important index of fish fertility determination of the sample’s ATP concentration was done by ultra sensitive bioluminescence method. ATP content of sperms were determined at two different sampling temperature (10-12 and 18-20°C) and two different keeping temperature (4°C and room temperature) for 6 h and also ATP content assayed until 10 days storage in the three extender types (glycerol, 0.7 and 0.65% salt solution). Results of the present study showed that, ATP content of sperms, collected at 10-12°C was 74.04±7.22%, in comparison to 18-20°C. The ATP content of sperms during 6 h keeping at 4°C and room temperature were 90.26±0.91% and 17.17±1.49%, respectively. Determination of sperms’ ATP content after 5 days keeping in glycerol, 0.7 and 0.65% salt solution revealed that glycerol or 0.65% salt solution is better extender than 0.7% salt solution. But sperms which were kept in mentioned extenders for 10 days showed that glycerol was better than salt solutions based on sperm ATP content saving. Results revealed that, in order to save sperms’ ATP content of golden grey mullet, sampling at 18-20°C and keeping in glycerol as extender is recommended.