Fiber Type-Specific Satellite Cell Content in Cyclists Following Heavy Training with Carbohydrate and Carbohydrate-Protein Supplementation

Abstract

The central purpose of this study was to evaluate the fiber type-specific satellite cell and myonuclear responses of endurance-trained cyclists to a block of intensified training, when supplementing with carbohydrate (CHO) vs. carbohydrate-protein (PRO). In a crossover design, endurance-trained cyclists (n = 8) performed two consecutive training periods, once supplementing with CHO (de facto "control" condition) and the other with PRO. Each training period consisted of 10 days of intensified cycle training (ICT-120% increase in average training duration) followed by 10 days of recovery (RVT-reduced volume training; 33% volume reduction vs. normal training). Skeletal muscle biopsies were obtained from the vastus lateralis before and after ICT and again following RVT. Immunofluorescent microscopy was used to quantify SCs (Pax7+), myonuclei (DAPI+), and myosin heavy chain I (MyHC I). Data are expressed as percent change ± 90% confidence limits. The 10-day block of ICT_CHO increased MyHC I SC content (35 ± 28%) and myonuclear density (16 ± 6%), which remained elevated following RVT_CHO (SC = 69 ± 50% vs. PRE; Nuclei = 17 ± 15% vs. PRE). MyHC II SC and myonuclei were not different following ICT_CHO, but were higher following RVT_CHO (SC = +33 ± 31% vs. PRE; Nuclei = 15 ± 14% vs. PRE), indicating a delayed response compared to MyHC I fibers. The MyHC I SC pool increased following ICT_PRO (37 ± 37%), but without a concomitant increase in myonuclei. There were no changes in MyHC II SC or myonuclei following ICT_PRO. Collectively, these trained endurance cyclists possessed a relatively large pool of SCs that facilitated rapid (MyHC I) and delayed (MyHC II) satellite cell proliferation and myonuclear accretion under carbohydrate conditions. The current findings strengthen the growing body of evidence demonstrating alterations in satellite cell number in the absence of hypertrophy. Satellite cell pool expansion is typically viewed as an advantageous response to exercise. However, when coupled with our previous report that PRO possibly enhanced whole muscle recovery and increased MyHC I and II fiber size, the limited satellite cell/myonuclear response observed with carbohydrate-protein seem to indicate that protein supplementation may have minimized the necessity for satellite cell involvement, thereby suggesting that protein may benefit skeletal muscle during periods of heavy training.

Keywords: endurance-trained cyclists; intensified training; myogenesis; protein supplementation.

Figure 1

General experimental design. Bx, Skeletal muscle biopsy.

Figure 2

Overlaid image of Pax7+/DAPI+ cells enclosed by laminin border. Image was captured at 20X magnification. (A) Laminin border of skeletal muscle fibers appearing purple with DAPI+ cells appearing blue. (B) Pax7+ structures appearing yellow denoted with white arrows. (C) DAPI+ cells appearing blue with white arrows denoting the myonuclei of Pax7+/DAPI+ cells. (D) Overlaid image with laminin, Pax7, and DAPI with white arrows denoting SCs that are Pax7+/DAPI+ and enclosed by a laminin border.