STORRE: Mixed muscle protein synthesis and …
C2C12 mouse myoblasts were purchased from ATCC (Manassas, VA). Cells were maintained in growth medium consisting of Dulbecco's modified eagle medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Cells were grown to confluence in a 37°C, 5% CO2 humidified environment. Once cells reached confluence, growth medium was removed, and the cells were rinsed with sterile phosphate-buffered saline. Differentiation medium consisting of Dulbecco's modified eagle medium supplemented with 2% horse serum, and 1% penicillin/streptomycin was added to all cells. For the assessment of protein synthesis, sterile 2H2O was added to differentiation media to a 4% isotopic enrichment. This enriched medium was added to cells either at the onset of differentiation and maintained for the full 7-day differentiation (7 days of isotope exposure), on the 3rd day of differentiation (4 days of isotope exposure), or on the 5th day of differentiation (2 days of isotope exposure). All cells were harvested on day 7 of the experiment. Cells were rinsed in phosphate-buffered saline and scraped in 1 ml of lysis buffer (100 mM KCl, 40 mM Tris·HCl, 10 mM Tris base, 5 mM MgCl2, 1 mM EDTA, 1 mM ATP, pH = 7.5) with phosphatase and protease inhibitors (HALT, Thermo Scientific, Rockford, IL).
protein synthesis Flashcards | Quizlet
Dr. Phillips: Anabolic resistance is, as we define it, the inability of skeletal muscle in aged persons to mount a full protein synthetic response similar to that seen in the young. In other words, older people just don’t put the protein they eat into their muscles as efficiently as young people. That means as we age our muscles gradually begin to make less protein so our muscle mass declines, otherwise called sarcopenia.
Three men and 3 women (27.8 ± 2.0 yr, 22.2 ± 1.0 body mass index (BMI), 56.1 ± 4.5 kg lean body mass (LBM)) received a 14-hour primed, constant intravenous infusion of L(1- 13 C)leucine to evaluate the fractional rate of mixed muscle protein synthesis.
to assess muscle protein synthesis and ..
4. Tang JE, Moore DR, Kujbida GW, Tarnopolsky MA and Phillips SM. Ingestion of whey hydrolysate, casein, or soy protein isolate: effects on mixed muscle protein synthesis at rest and following resistance exercise in young men. J Appl Physiol 107: 987-992, 2009.
Mechanical Ventilation Depresses Protein Synthesis in …
5. Moore DR, Robinson MJ, Fry JL, Tang JE, Glover EI, Wilkinson SB, Prior T, Tarnopolsky MA and Phillips SM. Ingested protein dose response of muscle and albumin protein synthesis after resistance exercise in young men. Am J Clin Nutr 89: 161-168, 2009.
in the rate of mixed muscle protein synthesis ..
Users of the flooding dose technique advocated for this method because of its relatively quick measurement compared with the 4- to 6-h primed continuous infusion, as well as the need for only a single biopsy of the tissue of interest. Indeed, because of this relative speed, a hybrid technique with decreased isotope concentration used as a flooding prime with the normal continuous infusion was recently proposed (). The shortened measurement period has advantages in that it decreases subject burden and can capture relatively acute changes in protein synthesis. However, as discussed below, there are disadvantages to shortening the time period of the continuous infusion in that it perhaps biases synthetic outcomes of a tissue sample. This concern is apparent when considering tissue protein heterogeneity.
Acute Post-Exercise Myofibrillar Protein Synthesis Is …
Compared with our previous report in the postabsorptive state, amino acid infusion increased the fractional rate of mixed muscle protein synthesis by 48% ( P