Differential response of oxidative and glycolytic skeletal muscle fibers to mesterolone

Abstract

Oxidative and glycolytic muscle fibers differ in their ultrastructure, metabolism, and responses to physiological stimuli and pathological insults. We examined whether these fibers respond differentially to exogenous anabolic androgenic steroids (AASs) by comparing morphological and histological changes between the oxidative anterior latissimus dorsi (ALD) and glycolytic pectoralis major (PM) fibers in adult avian muscles. Adult female White Leghorn chickens (Gallus gallus) were randomly divided into five groups: a vehicle control and four mesterolone treatment groups (4, 8, 12, and 16 mg/kg). Mesterolone was administered orally every three days for four weeks. Immunocytochemical techniques and morphometric analyses were employed to measure the changes in muscle weight, fiber size, satellite cell (SC) composition, and number of myonuclei. Mesterolone increased both body and muscle weights and induced hypertrophy in glycolytic PM fibers but not in oxidative ALD fibers. Mesterolone induced SC proliferation in both muscles; however, the myonuclear accretion was noticeable only in the PM muscle. In both muscles, the collective changes maintained a constant myonuclear domain size and the changes were dose independent. In conclusion, mesterolone induced distinct dose-independent effects in avian oxidative and glycolytic skeletal muscle fibers; these findings might be clinically valuable in the treatment of age-related sarcopenia.

Figure 1

Body and muscle weights in control and mesterolone-treated groups. C: control group; M1, M2, M3, M4: groups treated with 4, 8, 12, and 16 mg/kg mesterolone, respectively. ALD: anterior latissimus dorsi muscle; PM: pectoralis major muscle; SD: standard deviation. Different alphabets (a, b) indicate significant difference among the groups (ANOVA, LSD post-hoc).

Figure 2

Immunofluorescent identification of satellite cell nuclei (SCN) in cross sections of anterior latissimus dorsi muscles obtained from control and mesterolone-treated chickens. (A) and (B) represent a section obtained from a control bird. (C) and (D) represent a section obtained from a mesterolone-treated bird. (A) and (C) show all nuclei in blue (DAPI staining) and the basal laminae of muscle fibers in red (anti-laminin staining). (B) and (D) show SCN in green (anti-Pax7 labeling) and the basal laminae in red. Scale bar = 30 μm.

Figure 3

Immunofluorescent identification of satellite cell nuclei (SCN) in cross sections of pectoralis major muscles obtained from control and mesterolone-treated chickens. (A) and (B) represent a section obtained from a control bird. (C) and (D) represent a section obtained from a mesterolone-treated bird. (A) and (C) show all nuclei in blue (DAPI staining) and the basal laminae of muscle fibers in red (anti-laminin staining). (B) and (D) show SCN in green (anti-Pax7 labeling) and the basal laminae in red. As a unique histological feature of the chicken pectoralis, myonuclei are normally found deep within the fiber sarcoplasm. Scale bar = 30 μm.