The temperature and oxygen environment play important roles in the desorption of 137Cs from freshwater lake sediment to lake water. In this study, 12 quarterly surveys were performed to measure the dissolved 137Cs concentration in surface and bottom lake water, the vertical distribution of water temperature, and the dissolved oxygen (DO) concentration at the upstream, midstream, and downstream sites of the Yokokawa Dam Lake in Fukushima Prefecture, Japan. Higher concentrations of dissolved 137Cs were detected in the bottom water than in the surface water, especially in the summer and midstream lake regions at depths of 8–21 m owing to higher temperatures, which activated the bacterial decomposition of organic matter, and anaerobization, which enhanced the NH4+ in the pore water and 137Cs desorption from mineral particles. To compare the effects of anaerobization and increasing temperature on 137Cs desorption from sediment particles, intact sediment core samples were collected from the lake midstream and incubated for 1–14 days in a chamber under three controlled temperature and oxygen environment conditions: aerobic +10 °C; anaerobic +10 °C; and anaerobic +20 °C. The vertical distribution of 137Cs in the sediment pore water showed a similar profile as NH4+ and K+, and both the increased temperature and reduced DO concentration enhanced the 137Cs desorption. A comparison of the standard partial regression coefficients of temperature and DO concentration in the multiple regression equation for the 137Cs concentration in pore water shows that the reduction of DO from saturation to zero at the water–sediment interface accelerated the desorption of 137Cs more strongly than did the temperature rise from 10 to 20 °C. The experimental results show a nearly inverse proportional relationship between NH4+ and the distribution coefficients of 137Cs after thermodynamic correction, except in the surface layer. These findings indicate that the 137Cs concentration in pore water can be explained by the temperature-dependent chemical distribution between the solid–aqueous phase and its equilibrium with NH4+.