Acute and long-term hormonal and neuromuscular adaptations to hypertrophic strength training were studied in 13 recreationally strength-trained men. The experimental design comprised a 6-month hypertrophic strength-training period including 2 separate 3-month training periods with the crossover design, a training protocol of short rest (SR, 2 minutes) as compared with long rest (LR, 5 minutes) between the sets. Basal hormonal concentrations of serum total testosterone (T), free testosterone (FT), and cortisol (C), maximal isometric strength of the leg extensors, right leg 1 repetition maximum (1RM), dietary analysis, and muscle cross-sectional area (CSA) of the quadriceps femoris by magnetic resonance imaging (MRI) were measured at months 0, 3, and 6. The 2 hypertrophic training protocols used in training for the leg extensors (leg presses and squats with 10RM sets) were also examined in the laboratory conditions at months 0, 3, and 6. The exercise protocols were similar with regard to the total volume of work (loads 3 sets 3 reps), but differed with regard to the intensity and the length of rest between the sets (higher intensity and longer rest of 5 minutes vs. somewhat lower intensity but shorter rest of 2 minutes). Before and immediately after the protocols, maximal isometric force and electro-myographic (EMG) activity of the leg extensors were measured and blood samples were drawn for determination of serum T, FT, C, and growth hormone (GH) concentrations and blood lactate. Both protocols before the experimental training period (month 0) led to large acute increases (p < 0.05–0.001) in serum T, FT, C < and GH concentrations, as well as to large acute decreases (p < 0.05–0.001) in maximal isometric force and EMG activity. However, no significant differences were observed between the protocols. Significant increases of 7% in maximal isometric force, 16% in the right leg 1RM, and 4% in the muscle CSA of the quadriceps femoris were observed during the 6-month strength-training period. However, both 3-month training periods performed with either the longer or the shorter rest periods between the sets resulted in similar gains in muscle mass and strength. No statistically significant changes were observed in basal hormone concentrations or in the profiles of acute hormonal responses during the entire 6-month experimental training period. The present study indicated that, within typical hypertrophic strength-training protocols used in the present study, the length of the recovery times between the sets (2 vs. 5 minutes) did not have an influence on the magnitude of acute hormonal and neuromuscular responses or long-term training adaptations in muscle strength and mass in previously strength-trained men.