Objective To explore the distribution characteristics of neuron-specific enolase（NSE）, synaptophysin（SYN）and Neurofilament protein（NF）proteins in the developmental stages of human embryonic heart tissues. Methods Immunohistochemistry was used to detect the expressions of NSE, SYN and NF proteins in the heart tissues during the second, third and fourth month of human embryonic and fetal development(16 cases). Results In the embryos at the 2nd, 3rd and 4th months of gestation, NSE, SYN and NF proteins were positively expressed in the heart tissue. With the increase of gestational age, the intensities of the positive expression of NSE, SYN and NF in the heart tissues gradually reduced. At the 2nd month of gestation, the intensities of NSE, SYN and NF protein positive expressions were 86.79±7.75，133.03±13.61 and 114.32±11.12, respectively. At the 3rd month of gestation, the intensities of NSE, SYN and NF proteins positive expressions were 81.89±9.62，119.91±11.70 and 93.13±13.63, respectively. At the 4 month of gestation, NSE, SYN and NF proteins positive expression intensity values were 72.18±11.97，107.02±10.89 and 91.17±13.81, respectively.One-Way ANOVA andLSD-t test were employed to compare positive expression intensity values of NSE, SYN and NF proteins in heart tissues detected in the 3 fetal periods, and the results of NSE, SYN and NF showed significant difference in the heart tissue(P<0.05). Conclusion At the second to fourth gestation, NSE, NF and SYN proteins show specific distribution patterns in human embryonic cardiac muscle tissues. The expression intensities of NF, NSE and SYN in myocardium increases gradually with the increase of gestational age.