Effect of estrogen on apoptotic cell death was studied in the two sexually dimorphic neuronal groups in the developing rat preoptic area (POA): the anteroventral periventricular nucleus of the POA (AVPvN-POA); and the sexually dimorphic nucleus of the POA (SDN-POA). A specific labelling of nuclear fragmentation was performed by terminal deoxynucleotydyl transferase(TdT)-mediated dUTP-biotin nick end-labeling method (TUNEL method) to demonstrate apoptosis. In the AVPvN-POA whose size is larger in females than in males, the number of TUNEL-positive cells was not significantly different between day 5 control and female pups sacrificed 10 h after 25 micrograms estradiol benzoate (EB) injection. However, TUNEL-positive cells showed a significant increase in the female pups sacrificed 24 h after EB injection, compared to that shown in the control female pups. In the SDN-POA whose size is larger in males than in females, EB injection significantly decreased TUNEL-positive cells in the female pups sacrificed 24 h after EB injection, compared to that in controls. These results suggest that estrogen regulates the neuronal number by facilitating apoptotic cell death in the developing AVPvN-POA or by inhibiting it in the developing SDN-POA.