Helium-Neon laser puncture exposure on the reproductive acupoint causes generalized reproductive acceleration of female catfish broodstock. The purpose of this study was to evaluate the effect of laser puncture exposure and varied protein diets on the vitellogenin level and GSI (Gonado Somatic Index). We used 172 female mature catfish with ages of 1-1.5 years old. The research treatment were conducted using three protein levels diets (30%, 35% and 40%) and given to the female catfish broodstock with and without laser puncture treatment. Each fish was received laser puncture exposure on 2/3 ventral part of the body for 15 sec/week in 8 weeks. Blood and gonad were isolated from 4 samples per treatment group in each week. Vitellogenin level was analyzed by ELISA. The results showed that induction with laser puncture and varied protein levels diets were significantly increased the vitellogenin level (P < 0.001) and GSI number (P < 0.05) compared to the control fish. Laser puncture exposure and 30% protein dietary treatment at third week were more effective to increase the vitellogenin level and GSI. This study suggested that laser puncture induction along with 30% of protein level diets were accelerated the gonad maturation 3 weeks faster than control fish.