Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

New insights into skeletal muscle fibre types in the dog with particular focus towards hybrid myosin phenotypes

  • 419 Accesses

  • 40 Citations

Abstract

Electrophoresis, immunoblots, immunohistochemistry and image analysis methods were applied to characterise canine trunk and appendicular muscle fibres according to their myosin heavy chain (MyHC) composition and to determine, on a fibre-to-fibre basis, the correlation between contractile [MyHC (s), myofibrillar ATPase (mATPase) and sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) isoforms], metabolic [succinate dehydrogenase (SDH) and glycerol-3-phosphate dehydrogenase (GPDH) activities and glycogen and phospholamban (PLB) content] and morphological (cross-sectional area and capillary and nuclear densities) features of individual myofibres. An accurate delineation of MyHC-based fibre types was obtained with the developed immunohistochemical method, which showed high sensitivity and objectivity to delineate hybrid fibres with overwhelming dominance of one MyHC isoform. Phenotypic differences in contractile, metabolic and morphological properties seen between fibre types were related to MyHC content. All canine skeletal muscle fibre types had a relatively high histochemical SDH activity but significant differences existed in the order IIA>I>IIX. Mean GPDH was ranked according to fibre type such that I<IIA<IIX. Type IIA fibres were the smallest, type IIX fibres the largest and type I of intermediate size. Capillary and nuclear density decreased in the order IIA>I>IIX. Hybrid fibres, which represented nearly one third of the whole pool of skeletal muscle fibres analysed, had mean values intermediate between their respective pure phenotypes. Slow fibres expressed the slow SERCA isoform and PLB, whereas type II fibres expressed the fast SERCA isoform. Discrimination of myofibres according to their MyHC content was possible on the basis of their contractile, metabolic and morphological features. These intrafibre interrelationships suggest that myofibres of control dogs exhibit a high degree of co-ordination in their physiological, biochemical and morphological characteristics. This study demonstrates that canine skeletal muscle fibres have been misclassified in numerous previous studies and offers useful baseline data and new prospects for future work on muscle-fibre-typing in canine experimental studies.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

References

  1. Amann JF, Wharton RE, Madsen RW, Laughlin MH (1993) Comparison of muscle fiber types and oxidative capacity in gracilis, rectus femoris, and triceps brachii muscles in the ferret (Mustela putorius furo) and the domestic dog (Canis familiaris). Anat Rec 236:611–618

  2. Andersen P, Henriksson J (1977) Capillary supply of the quadriceps muscle of man: adaptive response to exercise. J Physiol (Lond) 270:677–690

  3. Argüello A, López–Fernández JL, Rivero JLL (2001) Limb myosin heavy chain isoproteins and muscle fiber types in the adult goat (Capra hircus). Anat Rec 264:184–293

  4. Blanco CE, Sieck GC, Edgerton VR (1988) Quantitative histochemistry determination of succinic dehydrogenase activity in skeletal muscle fibers. Histochem J 24:431–444

  5. Booth FW, Baldwin KM (1996) Muscle plasticity: energy demand and supply processes. In: Rowell LB, Shepherd JT (eds) Handbook of physiology. American Physiological Society, Bethesda, MD, pp 1075–1123

  6. Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  7. Braund KG, Hoff EJ, Richardson EY (1978) Histochemical identification of fiber types in canine skeletal muscle. Am J Vet Res 39:561–565

  8. Brooke MH, Kaiser KK (1970) Three “myosin adenosine triphosphatase” systems: the nature of their pH lability and sulfhydryl dependence. J Histochem Cytochem 18:670–672

  9. Cardinet GH III, Guffy MM, Wallace LJ, Laben RC (1983) Canine hip dysplasia in German shepherd dog-greyhound cross-breeds. J A Vet Med Assoc 182:393–395

  10. Castle ME, Reyman TA (1984) The effect of tenotomy and tendon transfers on muscle fiber types in the dog. Clin Orthop 186:302–310

  11. Chikuni K, Muroya S, Nakajima I (2004) Absence of the functional myosin heavy chain 2b isoform in equine skeletal muscles. Zool Sci 21:589–596

  12. Christensen L, Strange L (1987) Universal immunoperoxidase staining protocol to optimize the use of polyclonal and monoclonal antibodies. J Histotechnol 10:11–15

  13. Delp MD, Duan C (1996) Composition and size of type I, IIA, IID/X, and IIB fibers and citrate synthase activity of rat muscle. J Appl Physiol 80:261–270

  14. Duris M-P, Picard B, Geay Y (2002) Specificity of different anti-myosin heavy chain antibodies in bovine muscle. Meat Sci 55:67–78

  15. Eizema K, Burg M van den, Kiri A, Dingboom EG, Oudheusden H van, Goldspink G, Weijs WA (2003) Differential expression of equine myosin heavy-chain mRNA and protein isoforms in a limb muscle. J Histochem Cytochem 51:1207–1216

  16. Gorza L (1990) Identification of a novel type 2 fiber population in mammalian skeletal muscle by combined use of histochemical myosin ATPase and anti-myosin monoclonal antibodies. J Histochem Cytochem 38:257–265

  17. Graziotti GH, Ríos CM, Rivero JLL (2001) Evidence for three fast myosin heavy chain isoforms in type II skeletal muscle fibers in adult llama (Lama glama). J Histochem Cytochem 49:1033–1044

  18. Graziotti GH, Palencia P, Delhon G, Rivero JLL (2004) Neuromuscular partitioning, architectural design, and myosin fiber types of the M. vastus lateralis of the llama (Lama glama). J Morphol 262:667–681

  19. Green HJ, Reichmann H, Pette D (1982) A comparison of two ATPase based schemes for histochemical muscle fibre typing in various mammals. Histochemistry 76:21–31

  20. Guth L, Samaha FJ (1969) Procedure for the histochemical demonstration of actomyosin ATPase. Exp Neurol 28:365–366

  21. Hämäläinen N, Pette D (1993) The histochemical profiles of fast fiber types IIB, IID, and IIA skeletal muscles of mouse, rat, and rabbit. J Histochem Cytochem 41:733–743

  22. Hämäläinen N, Pette D (1997) Co-ordinated fast-to-slow transitions of myosin and SERCA isoforms in chronically stimulated muscles of euthyroid and hyperthyroid rabbits. J Muscle Res Cell Motil 18:545–554

  23. Horton MJ, Brandon CA, Morris TJ, Braun TW, Yaw KM, Sciote JJ (2001) Abundant expression of myosin heavy-chain IIB RNA in a subset of human masseter muscle fibers. Arch Oral Biol 46:1039–1050

  24. Hu P, Zhang KM, Free JJ, Wang SW, Wright LD, Wechsler AS, Spratt JA, Briggs FN (1996) Salbutamol and chronic low-frequency stimulation of canine skeletal muscle. J Physiol (Lond) 496:221–227

  25. Jorgensen AO, Arnold W, Pepper DR, Kahl SD, Mandel F, Campbell KP (1988) A monoclonal antibody to the Ca2+-ATPase of cardiac sarcoplasmic reticulum cross-reacts with slow type I but not with fast type II canine skeketal muscle fibres: an immynocytochemical and immunochemical study. Cell Motil Cytoskelet 9:164–174

  26. Latorre R, Gil F, Vázquez JM, Moreno F, Mascarello F, Ramírez G (1993) Skeletal muscle fiber types in the dog. J Anat 182:329–337

  27. Lefaucheur L, Ecolan P, Plantard L, Gueguen N (2002) New insights into muscle fiber types in the pig. J Histochem Cytochem 50:719–730

  28. Lucas CA, Kang LHD, Hoh JFY (2000) Monospecific antibodies against the three mammalian fast limb myosin heavy chains. Biochem Biophys Res Comm 272:303–308

  29. Lytton J, Westlin M, Burk SE, Shull GE, Maclennan DH (1992) Functional comparisons between isoforms of the sarcoplasmic or endoplasmic reticulum family of calcium pumps. J Biol Chem 267:14483–14489

  30. Mabuchi K, Sreter FA, Gergely J, Jorgensen AO (1990) Myosin and sarcoplasmic reticulum Ca2+-ATPase isoforms in electrically stimulated rabbit fast muscle. In: Pette D (ed) The dynamic state of muscle fibres. De Gruyter, Berlin, pp 445–462

  31. Maccatrozzo L, Patruno M, Toniolo L, Reggiani C, Mascarello F (2004) Myosin heavy chain 2B isoform is expressed in specialized eye muscles but not in trunk and limb muscles of cattle. Eur J Histochem 48:357–366

  32. Martin TP, Vailas AC, Durivage JB, Edgerton VR, Castleman KR (1985) Quantitative histochemical determination of muscle enzymes: biochemical verification. J Histochem Cytochem 33:1053–1059

  33. Nibbering PH, Marijnene JGJ, Raap AK, Leijh PCJ, Furth R van (1986) Quantitative study enzyme immunocytochemical reactions performed with enzyme conjugates immobilized on nitrocellulose. Histochemistry 84:538–543

  34. Nwoye L, Mommaerts WFHM, Simpson DR, Sreyderian K, Marushi M (1982) Evidence for a direct action of thyroid hormone in specifying muscle properties. Am J Physiol 242:R401–R408

  35. Pellegrino MA, Canepari M, Rossi R, D'Antona G, Reggiani G, Bottinelli R (2003) Orthologous myosin isoforms and scaling of shortening velocity with body size in mouse, rat, rabbit and human muscles. J Physiol (Lond) 543:677–689

  36. Pette D, Staron RS (2000) Myosin isoforms, muscle fiber types, and transitions. Microsc Res Tech 50:500–509

  37. Picard B, Lefaucheur L, Fauconneau B, Rémignon H, Cherel Y, Barrey E, Nedelec J (1998) Dossier: caracterisation des differents types de fibers musculaires dans plusieurs espèces, production et utilisation d'anticorps monoclonaux diriges contre les chaînes lourdes de myosine rapide IIa et IIb. INRA Prod Anim 11:145–163

  38. Quiroz-Rothe E, Rivero JLL (2001) Co-ordinated expression of contractile and non-contractile features of control equine muscle fiber types characterized by immunostaining of myosin heavy chains. Histochem Cell Biol 116:299–312

  39. Quiroz-Rothe E, Rivero JLL (2004) Coordinated expression of myosin heavy chains, metabolic enzymes, and morphological features of porcine skeletal muscle fiber types. Microsc Res Tech 65:43–61

  40. Reichmann H, Pette D (1982) A comparative microphotometric study of succinate dehydrogenase activity levels in type I, IIA and IIB fibres of mammalian and human muscles. Histochemistry 74:27–41

  41. Rivero JLL, Diz A, Toledo M, Agüera E (1994) Enzyme-histochemical profiles of fiber types in mature canine appendicular muscles. Anat Histol Embryol 23:330–336

  42. Rivero JLL, Talmadge RJ, Edgerton VR (1996) Correlation between myofibrillar ATPase activity and myosin heavy chain composition in equine skeletal muscle and the influence of training. Anat Rec 246:195–207

  43. Rivero JLL, Serrano AL, Barrey E, Valette JP, Jouglin M (1999a) Analysis of myosin heavy chains at the protein level in horse skeletal muscle. J Muscle Res Cell Motil 20:211–221

  44. Rivero JLL, Talmadge RJ, Edgerton VR (1999b) Interrelationships of myofibrillar ATPase activity and metabolic properties of myosin heavy chain-based fiber types in rat skeletal muscle. Histochem Cell Biol 111:277–287

  45. Rossini K, Rizzi C, Sandri M, Bruson A, Carraro U (1995) High-resolution sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunohistochemical identification of the 2X and embryonic myosin heavy chains in complex mixtures of isomyosins. Electrophoresis 16:101–104

  46. Roy RR, Monke SR, Allen DL, Edgerton VR (1999) Modulation of myonuclear number in functionally overloaded and exercised rat plantaris fibers. J Appl Physiol 87:634–642

  47. Sant'Ana-Pereira JAA, Wessels A, Nijtmans L, Moorman AFM, Sargeant AJ (1995) New method for the accurate characterization of single human skeletal muscle fibres demonstrates a relation between mATPase and MyHC expression in pure and hybrid fibre types. J Muscle Res Cell Motil 16:21–34

  48. Schiaffino S, Reggiani C (1996) Molecular diversity of myofibrillar proteins: gene regulation and functional significance. Physiol Rev 76:271–423

  49. Schiaffino S, Salviati G (1998) Molecular diversity of myofibrillar proteins: isoform analysis at the protein and mRNA level. Methods Cell Biol 52:349–369

  50. Schiaffino S, Gorza L, Sartore S, Saggin L, Ausoni S, Vianello M, Gundersen K, Lomo T (1989) Three myosin heavy chain isoforms in type 2 skeletal muscle fibers. J Muscle Res Cell Motil 10:197–205

  51. Sciote JJ, Rowlerson A (1998) Skeletal fiber types and spindle distribution in limb and jaw muscles of the adult neonatal opossum, Monodelphis domestica. Anat Rec 251:548–562

  52. Serrano AL, Petrie JL, Rivero JLL, Hermanson JW (1996) Myosin isoforms and muscle fiber characteristics in equine gluteus medius muscle. Anat Rec 244:444–451

  53. Serrano AL, Pérez M, Lucía A, Chicharro JL, Quiroz-Rothe E, Rivero JLL (2001) Immunolabelling, histochemistry and in situ hybridisation in human skeletal muscle fibres to detect myosin heavy chain expression at the protein and mRNA level. J Anat 199:329–337

  54. Shrager JB, Desjardins PR, Burkan JM, Konig SK, Steart SK, Su L, Shah MC, Bricklin E, Tewari M, Hoffman R, Rickel MR, Jullian EH, Rubinstein NA, Stedman HH (2000) Human skeletal myosin heavy chain genes are tightly linked in the order embryonic-IIa-IId/x-IIb-perinatal-extraocular. J Muscle Res Cell Motil 21:345–355

  55. Sieck GC, Zhan W, Prakash YS, Daood MJ, Watchko JF (1995) SDH and actomyosin ATPase activities of different fibre types in rat diaphragm muscle. J Appl Physiol 79:1629–1639

  56. Simmerman HKB, Jones LR (1998) Phospholamban: protein structure, mechanism of action, and role in cardiac function. Physiol Rev 78:921–947

  57. Smerdu V, Karsh-Mizrachi I, Campione M, Leinwadn L, Schiaffino S (1994) Type IIx myosin heavy chain transcripts are expressed in type IIb fibrers of human skeletal muscle. Am J Physol 267:C1723–C1728

  58. Snow DH, Billeter R, Mascarello F, Carpene E, Rowlerson A, Jenny E (1982) No classical IIB fibres in dog skeletal muscle. Histochemistry 75:53–65

  59. Staron RS (1991) Correlation between myofibrillar ATPase activity and myosin heavy chain composition in single human muscle fibres. Histochemistry 96:21–24

  60. Stephenson GMM (2001) Hybrid skeletal muscle fibres: a rare or common phenomenon? Clin Exp Pharmacol Physiol 28:692–702

  61. Strbenc M, Smerdu V, Zupanc M, Tozon N, Fazarinc G (2004) Pattern of myosin heavy chain isoforms in different fibre types of canine trunk and limb skeletal muscles. Cell Tiss Org 176:178–186

  62. Talmadge RJ, Roy RR (1993) Electrophoretic separation of rat skeletal muscle myosin heavy-chain isoforms. J Appl Physiol 75:2337–2340

  63. Talmadge RJ, Roy RR, Edgerton VR (1993) Muscle fiber types and function. Curr Opin Rheumatol 5:695–705

  64. Talmadge RJ, Roy RR, Chalmers GR, Edgerton VR (1996a) MHC and sarcoplasmic reticulum protein isoforms in functionally overloaded cat plantaris muscle fibers. J Appl Physiol 80:1296–1303

  65. Talmadge RJ, Grossman EJ, Roy RR (1996b) Myosin heavy chain composition of adult feline (Felis catus) limb and diaphragm muscles. J Exp Zool 275:413–420

  66. Tanabe R, Muroya S, Chikuni K (1998) Sequencing of the 2a, 2x, and slow isoforms of the bovine myosin heavy chain and the different expression among muscles. Mamm Genome 9:1056–1058

  67. Toniolo L, Patrono M, Maccatrozzo L, Pellegrino MA, Canepari M, Rossi R, D'Antona G, Bottinelli R, Reggiani C, Mascarello F (2004) Fast fibres in a large animal: fibre types, contractile properties and myosin expression in pig skeletal muscles. J Exp Biol 207:1875–1886

  68. Towbin H, Staehlin T, Gordeon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci 184:441–450

  69. Tseng BS, Kasper CE, Edgerton VR (1994) Cytoplasm-to-myonucleus ratios and succinate dehydrogenase activities in adult rat slow and fast muscle fibers. Cell Tissue Res 275:39–49

  70. Weiss A, McDonough D, Wertman B, Acakpo-Satchivi L, Montgomery K, Kucherlapati R, Leinwand L, Krauter K (1999) Organization of human and mouse skeletal myosin heavy chain gene clusters is highly conserved. Proc Natl Acad Sci U S A 16:2958–2963

  71. Wu YZ, Baker MJ, Crumley RL, Blanks RH, Caiozzo VJ (1998) A new concept in laryngeal muscle: multiple myosin isoform types in single muscle fibers of the lateral cricoarytenoid. Otolaryngol Head Neck Surg 118:86–94

  72. Wu YZ, Crumley RL, Caiozzo VJ (2000) Are hybrid fibers a common motif of canine laryngeal muscles? Single-fiber analyses of myosin heavy-chain isoform composition. Arch Otolaryngol Head Neck Surg 126:865–873

  73. Zhang KM, Hu P, Wang SW, Feher JJ, Wright LD, Weschler AS, Spratt JA, Brigss FN (1996) Salbutamol changes the molecular and mechanical properties of canine skeletal muscle. J Physiol (Lond) 496:211–220

Download references

Acknowledgements

We thank Prof. Stefano Schiaffino (University of Padova, Italy) for his generous gift of antibodies. Antibody S5-8H2 was a generous gift of Dr. Eric Barrey (INRA, France).

Author information

Correspondence to José-Luis L. Rivero.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Acevedo, L.M., Rivero, J.L. New insights into skeletal muscle fibre types in the dog with particular focus towards hybrid myosin phenotypes. Cell Tissue Res 323, 283–303 (2006). https://doi.org/10.1007/s00441-005-0057-4

Download citation

Keywords

  • Myosin heavy chain
  • Monoclonal antibodies
  • Immunohistochemistry
  • Myonuclear domain size
  • SERCA isoforms
  • Dog