It is commonly assumed that most (>95%) of the mercury (Hg) found in fish muscle is the toxic form, methylmercury (MeHg), due to its efficient assimilation and retention in biotic tissue. However, this assumption is largely based on studies examining the percentage of MeHg (%MeHg [the fraction of total Hg as MeHg]) in muscle from mostly large-bodied predatory fish; less is known about the %MeHg in smaller bodied individuals or those of different trophic guilds. The present study analyzed MeHg and total Hg concentrations in the muscle of 2 large-bodied piscivores (walleye and northern pike), one large-bodied benthivore (white sucker), and 2 small-bodied forage fish (sculpins and shiners) across a broad size range. We found substantially lower %MeHg than the commonly assumed 95% in several fish (e.g., 17 individuals had <70% MeHg). Muscle %MeHg significantly increased with size and age in all species except walleye, which had significantly higher %MeHg than pike or suckers, particularly in smaller and younger fish (e.g., 18-21% higher at 10 g; 5-11% higher at 500 g). Results of predictive modeling suggest that muscle %MeHg is higher in pelagic-feeding fish and those with lower lipid content, although model results varied significantly among species. According to our findings, total Hg measurement in muscle is not an appropriate proxy for MeHg in smaller fish from all species, an important consideration for future piscine Hg studies and monitoring. Environ Toxicol Chem 2018;37:2682-2691. © 2018 SETAC.