Abstract
The secretion of proteins labelled by incorporation of radioactive amino acids was studied in innervated and 10 to 13-day-denervated mouse skeletal muscle. The secretion of 3H-leucine-labelled proteins, expressed per mg muscle wet weight, increased after denervation, and the kinetics of the secretory process was also altered in denervated muscle. Separation of secreted 35S-methionine-labelled proteins by sodium dodecyl sulphate polyacrylamide gel electrophoresis followed by autoradiography revealed some denervation-induced alterations in the pattern of secreted proteins. The secretion from both innervated and denervated muscle was highly temperature sensitive and was reversibly inhibited by brefeldin A, a drug that blocks forward membrane transport from the endoplasmic reticulum/Golgi apparatus. This drug was also found to inhibit the uptake of fluorescein isothiocyanate-labelled dextran in denervated muscle but had no effect on the endocytotic activity of innervated muscle. This lends support to the hypothesis that the increased endocytotic activity in denervated muscle is coupled to a high secretory activity.
Similar content being viewed by others
Abbreviations
- BF A :
-
Brefeldin A
- dpm :
-
Disintegrations per minute
- EDL :
-
extensor digitorum longus
- FITC :
-
fluorescein isothiocyanate
- LDH :
-
lactate dehydrogenase
- SDS-P AGE :
-
sodium dodecyl sulphate-polyacrylamide gel electrophoresis
- TCA :
-
trichloroacetic acid
References
Bergmeyer HU (1974) Methods of Enzymatic Analysis, 2nd English edn, vol 2. Verlag Chemie, Academic Press, New York
Cooper DN, Barondes SH (1990) Evidence for export of a muscle lectin from cytosol to extracellular matrix and for a novel secretory mechanism. J Cell Biol 110:1681–1691
Covault J, Cunningham JM, Sanes JR (1987) Neurite outgrowth on cryostat sections of innervated and denervated skeletal muscle. J Cell Biol 105:2479–2488
Duve C de, Barsy T de, Poole B, Trouet A, Tulkens P, Hoof F van (1974) Lysosomotropic agents. Biochem Pharmacol 23:2495–2531
Elmquist S, Libelius R, Lawoko G, Tågerud S (1992) Dextrans as markers for endocytosis in innervated and denervated skeletal muscle. Muscle Nerve 15:876–884
Goldspink DF (1976) The effects of denervation on protein turnover of rat skeletal muscle. Biochem J 156:71–80
Henderson CE (1988) The role of muscle in the development and differentiation of spinal motoneurons: in vitro studies. In: Plasticity of the neuromuscular system. (Ciba Foudation Symposium 138) Wiley, Chichester, pp 172–191
Klausner RD, Donaldson JD, Lippincott-Schwartz J (1992) Brefeldin A: Insights into the control of membrane traffic and organelle structure. J Cell Biol 116:1071–1080
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Libelius R (1975) Evidence for endocytic uptake of cobra neurotoxin in mouse skeletal muscle. J Neural Transm 37:61–71
Libelius R, Tågerud S (1984) Uptake of horseradish peroxidase in denervated skeletal muscle occurs primarily at the endplate region. J Neurol Sci 66:273–281
Libelius R, Tågerud S (1989) Lysosomes in skeletal muscle. In: Sellin LC, Libelius R, Thesleff S (eds) Neuromuscular Junction. Fernström Foundation Series, Elsevier, pp 481–485
Libelius R, Lundquist I, Templeton W, Thesleff S (1978) Intracellular uptake and degradation of extracellular tracers in mouse skeletal muscle in vitro: The effect of denervation. Neuroscience 3:641–647
Libelius R, Josefsson J-O, Lundquist I (1979) Endocytosis in chronically denervated mouse skeletal muscle. A biochemical and ultrastructural study with horseradish peroxidase. Neuroscience 4:283–292
Mignatti P, Morimoto T, Rifkin DB (1992) Basic fibroblast growth factor, a protein devoid of secretory signal sequence, is released by cells via a pathway independent of the endoplasmic reticulum-Golgi complex. J Cell Physiol 151:81–93
Miller SG, Carnell L, Moore H-PH (1992) Post-Golgi membrane traffic: Brefeldin A inhibits export from distal Golgi compartments to the cell surface but not recycling. J Cell Biol 118:267–283
Misumi Y, Misumi Y, Miki K, Takatsuki A, Tamura G, Ikehara Y (1986) Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes. J Biol Chem 261:11398–11403
Rubartelli A, Sitia R (1991) Interleukin 1β and thioredoxin are secreted through a novel pathway of secretion. Biochem Soc Trans 19:255–259
Rubartelli A, Cozzolino F, Talio M, Sitia R (1990) A novel secretory pathway for interleukin-1β, a protein lacking a signal sequence. EMBO J 9:1503–1510
Slack JR, Hopkins WG, Pocket S (1983) Evidence for a motor nerve growth factor. Muscle Nerve 6:243–252
Takatsuki A, Tamura G (1985) Brefeldin A, a specific inhibitor of intracellular translocation of vesicular stomatitis G protein: Intracellular accumulation of high-mannose type G protein and inhibition of its cell surface expression. Agric Biol Chem 49:899–890
Thesleff S, Libelius R, Tågerud S (1989) Some aspects of long term regulations of nerve-muscle relations. In: Clifford Rose F, Jones R, Vrbova G (eds) Neuromuscular stimulation: Basic concepts and clinical applications. Demos Publications, New York, pp 27–36
Tågerud S, Libelius R (1984) Lysosomes in skeletal muscle following denervation. Time course of horseradish peroxidase uptake and increase of lysosomal enzymes. Cell Tissue Res 236:73–79
Tågerud S, Libelius R (1985) Receptor-mediated uptake of horseradish peroxidase in innervated and denervated skeletal muscle. Exp Cell Res 160:95–105
Tågerud S, Libelius R, Thesleff S (1986a) An ultrastructural study of the segmental uptake of horseradish peroxidase in the endplate region of denervated skeletal muscle fibres. J Neurol Sci 75:141–157
Tågerud S, Jirmanova I, Libelius R (1986b) Biochemical and ultrastructural effects of chloroquine on horseradish peroxidase uptake and lysosomal enzyme activities in innvervated and denervated mouse skeletal muscle. J Neurol Sci 75:159–171
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vult von Steyern, F., Kanje, M. & Tågerud, S. Protein secretion from mouse skeletal muscle: coupling of increased exocytotic and endocytotic activities in denervated muscle. Cell Tissue Res 274, 49–56 (1993). https://doi.org/10.1007/BF00327984
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00327984