Advertisement

Histochemistry and Cell Biology

, Volume 152, Issue 5, pp 323–331 | Cite as

3D analysis of capillary network in skeletal muscle of obese insulin-resistant mice

  • Nejc UmekEmail author
  • Simon Horvat
  • Erika Cvetko
  • Marko Kreft
  • Jiří Janáček
  • Lucie Kubínová
  • Tatjana Stopar Pintarič
  • Ida Eržen
Original Paper

Abstract

In obesity, the skeletal muscle capillary network regresses and the insulin-mediated capillary recruitment is impaired. However, it has been shown that in the early stage of advanced obesity, an increased functional vascular response can partially compensate for other mechanisms of insulin resistance. The present study aimed to investigate the changes in the capillary network around individual muscle fibres during the early stage of obesity and insulin resistance in mice using 3D analysis. Capillaries and muscle fibres of the gluteus maximus muscles of seven high-fat-diet-induced obese and insulin-resistant mice and seven age-matched lean healthy mice were immunofluorescently labelled in thick transverse muscle sections. Stacks of images were acquired using confocal microscope. Capillary network characteristics were estimated by methods of quantitative image analysis. Muscle fibre typing was performed by histochemical analysis of myosin heavy chain isoforms on thin serial sections of skeletal muscle. Capillary length per muscle fibre length and capillary length per muscle fibre surface were increased by 27% and 23%, respectively, around small muscle fibres in obese mice, while there were no significant comparative differences around large fibres of obese and lean mice. Furthermore, the capillarization was larger around small compared to large fibres and there was a shift toward fast type myosin heavy chain isoforms, with no significant changes in muscle fibre diameters, tortuosity and anisotropy in obese mice. Overall, the results show that obese insulin-resistant mice have selective increase in capillarization around small predominantly intermediate muscle fibres, which is most likely related to the impaired glucose metabolism characteristic of type 2 diabetes.

Keywords

Obesity Insulin resistance Capillary network 3D analysis Skeletal muscle Fibre type 

Notes

Acknowledgements

We are thankful to David Vondrášek, Nataša Pollak Kristl, Ivan Blažinovič, Majda Črnak Maasarani, Friderik Štendler, Marko Slak, Andreja Vidmar, Stanko Kristl, Milan Števanec, Vesna Mrak and Liljana Markova for technical support, and to Miha Pintarič and Chiedozie K. Ugwoke for manuscript proofreading.

Funding

This study was funded by the Slovenian Research Agency (Grant Nos: P3-0043, P4-0220 and P3-0310), the Czech Ministry of Education Youth and Sports (LM2015062 Czech-BioImaging), the European Regional Development Fund (OPPK BrainView CZ.2.16/3.1.00/21544), the Operational program for Research Development and Education (CZ.02.1.01/0.0/0.0/16_013/0001775 Modernization and support of research activities of the national infrastructure for biological and medical imaging Czech-BioImaging) and the University Medical Centre Ljubljana Tertiary Funding.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest, financial or otherwise regarding the publication of this paper.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. The Ethical Committee for laboratory animals of the Republic of Slovenia reviewed and approved all animal study protocols (Permit Number: U34401-21/2013/6).

References

  1. Ahmed SK, Egginton S, Jakeman PM, Mannion AF, Ross HF (1997) Is human skeletal muscle capillary supply modelled according to fibre size or fibre type? Exp Physiol 82:231–234CrossRefGoogle Scholar
  2. Andrikopoulos S, Blair AR, Deluca N, Fam BC, Proietto J (2008) Evaluating the glucose tolerance test in mice. Am J Physiol Endocrinol Metab 295:E1323–E1332.  https://doi.org/10.1152/ajpendo.90617.2008 CrossRefPubMedGoogle Scholar
  3. Baum O, Bigler M (2016) Pericapillary basement membrane thickening in human skeletal muscles. Am J Physiol Heart Circ Physiol 311:H654–H666.  https://doi.org/10.1152/ajpheart.00048.2016 CrossRefPubMedGoogle Scholar
  4. Blüher M (2010) The distinction of metabolically ‘healthy’ from ‘unhealthy’ obese individuals. Curr Opin Lipidol 21:38–43.  https://doi.org/10.1097/MOL.0b013e3283346ccc CrossRefPubMedGoogle Scholar
  5. Čebašek V, Eržen I, Vyhnal A, Janáček J, Ribarič S, Kubínová L (2010) The estimation error of skeletal muscle capillary supply is significantly reduced by 3D method. Microvasc Res 79:40–46.  https://doi.org/10.1016/j.mvr.2009.11.005 CrossRefPubMedGoogle Scholar
  6. Chadderdon SM, Belcik JT, Bader L, Peters DM, Kievit P, Alkayed NJ, Kaul S, Grove KL, Lindner JR (2016) Temporal changes in skeletal muscle capillary responses and endothelial-derived vasodilators in obesity-related insulin resistance. Diabetes 65:2249–2257.  https://doi.org/10.2337/db15-1574 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Charles JP, Cappellari O, Spence AJ, Hutchinson JR, Wells DJ (2016) Musculoskeletal geometry, muscle architecture and functional specialisations of the mouse hindlimb. PLoS One 11:e0147669.  https://doi.org/10.1371/journal.pone.0147669 CrossRefPubMedPubMedCentralGoogle Scholar
  8. Clerk LH, Vincent MA, Jahn LA, Liu Z, Lindner JR, Barrett EJ (2006) Obesity blunts insulin-mediated microvascular recruitment in human forearm muscle. Diabetes 55:1436–1442.  https://doi.org/10.2337/db05-1373 CrossRefPubMedGoogle Scholar
  9. Cvetko E, Karen P, Eržen I (2012) Myosin heavy chain composition of the human sternocleidomastoid muscle. Ann Anat 194:467–472.  https://doi.org/10.1016/j.aanat.2012.05.001 CrossRefPubMedGoogle Scholar
  10. Elias I, Franckhauser S, Bosch F (2013) New insights into adipose tissue VEGF-A actions in the control of obesity and insulin resistance. Adipocyte 2:109–112.  https://doi.org/10.4161/adip.22880 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Ellis CG, Goldman D, Hanson M, Stephenson AH, Milkovich S, Benlamri A, Ellsworth ML, Sprague RS (2010) Defects in oxygen supply to skeletal muscle of prediabetic ZDF rats. Am J Physiol Heart Circ Physiol 298:H1661–H1670.  https://doi.org/10.1152/ajpheart.01239.2009 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Engin A (2017) The definition and prevalence of obesity and metabolic syndrome. Adv Exp Med Biol 960:1–17.  https://doi.org/10.1007/978-3-319-48382-5_1 CrossRefPubMedGoogle Scholar
  13. Eriksson KF, Saltin B, Lindgarde F (1994) Increased skeletal muscle capillary density precedes diabetes development in men with impaired glucose tolerance: a 15-year follow-up. Diabetes 43:805–808.  https://doi.org/10.2337/diab.43.6.805 CrossRefPubMedGoogle Scholar
  14. Eržen I, Janáček J, Kubínová L (2011) Characterization of the capillary network in skeletal muscles from 3D data. Physiol Res 60:1–13PubMedGoogle Scholar
  15. Eržen I, Janáček J, Kreft M, Kubínová L, Cvetko E (2018) Capillary network morphometry of pig soleus muscle significantly changes in 24 hours after death. J Histochem Cytochem 66:23–31.  https://doi.org/10.1369/0022155417737061 CrossRefPubMedGoogle Scholar
  16. Gavin TP, Stallings HW, Zwetsloot KA, Westerkamp LM, Ryan NA, Moore RA, Pofahl WE, Hickner RC (2005) Lower capillary density but no difference in VEGF expression in obese vs. lean young skeletal muscle in humans. J Appl Physiol 98:315–321.  https://doi.org/10.1152/japplphysiol.00353.2004 CrossRefPubMedGoogle Scholar
  17. Gomes JLP, Fernandes T, Soci UPR, Silveira AC, Barretti DLM, Negrão CE, Oliveira EM (2017) Obesity downregulates microRNA-126 inducing capillary rarefaction in skeletal muscle: effects of aerobic exercise training. Oxid Med Cell Longev 2017:1–9.  https://doi.org/10.1155/2017/2415246 CrossRefGoogle Scholar
  18. Gute D, Laughlin MH, Amann JF (1994) Regional changes in capillary supply in skeletal muscle of interval-sprint and low-intensity, endurance-trained rats. Microcirculation 1:183–193CrossRefGoogle Scholar
  19. Harris BA (2005) The influence of endurance and resistance exercise on muscle capillarization in the elderly: a review. Acta Physiol Scand 185:89–97.  https://doi.org/10.1111/j.1365-201X.2005.01461.x CrossRefPubMedGoogle Scholar
  20. Hepple RT, Mathieu-Costello O (2017) Estimating the size of the capillary-to-fiber interface in skeletal muscle: a comparison of methods. J Appl Physiol 91:2150–2156.  https://doi.org/10.1152/jappl.2001.91.5.2150 CrossRefGoogle Scholar
  21. Hermansen L, Wachtlova M (1971) Capillary density of skeletal muscle in well-trained and untrained men. Acta Physiol Scand 30:860-863.  https://doi.org/10.1152/jappl.1971.30.6.860 CrossRefGoogle Scholar
  22. Janáček J, Čebašek V, Kubínová L, Ribarič S, Eržen I (2009) 3D visualization and measurement of capillaries supplying metabolically different fiber types in the rat extensor digitorum longus muscle during denervation and reinnervation. J Histochem Cytochem 57:437–447.  https://doi.org/10.1369/jhc.2008.953018 CrossRefPubMedPubMedCentralGoogle Scholar
  23. Janáček J, Cvetko E, Kubínová L, Travnik L, Eržen I (2011) A novel method for evaluation of capillarity in human skeletal muscles from confocal 3D images. Microvasc Res 81:231–238.  https://doi.org/10.1016/j.mvr.2010.11.012 CrossRefPubMedGoogle Scholar
  24. Janáček J, Kreft M, Čebašek V, Eržen I (2012) Correcting the axial shrinkage of skeletal muscle thick sections visualized by confocal microscopy. J Microsc 246:107–112.  https://doi.org/10.1111/j.1365-2818.2011.03594.x CrossRefPubMedGoogle Scholar
  25. Karen P, Stevanec M, Smerdu V, Cvetko E, Kubínová L, Erzen I (2009) Software for muscle fibre type classification and analysis. Eur J Histochem 53:87–95.  https://doi.org/10.4081/ejh.2009.e11 CrossRefPubMedGoogle Scholar
  26. Kim F, Pham M, Maloney E, Rizzo NO, Morton GJ, Wisse BE, Kirk EA, Chait A, Schwartz MW (2008) Vascular inflammation, insulin resistance, and reduced nitric oxide production precede the onset of peripheral insulin resistance. Arterioscler Thromb Vasc Biol 28:1982–1988.  https://doi.org/10.1161/ATVBAHA.108.169722 CrossRefPubMedPubMedCentralGoogle Scholar
  27. King AJ (2012) The use of animal models in diabetes research. Br J Pharmacol 166:877–894.  https://doi.org/10.1111/j.1476-5381.2012.01911.x CrossRefPubMedPubMedCentralGoogle Scholar
  28. Kondo H, Fujino H, Murakami S, Nagatomo F, Roy RR, Ishihara A (2011) Regressed three-dimensional capillary network and inhibited angiogenic factors in the soleus muscle of non-obese rats with type 2 diabetes. Nutr Metab (Lond) 8:77.  https://doi.org/10.1186/1743-7075-8-77 CrossRefGoogle Scholar
  29. Krogh A (1919) The number and distribution of capillaries in muscles with calculations of the oxygen pressure head necessary for supplying the tissue. J Physiol 52:409–415.  https://doi.org/10.1113/jphysiol.1919.sp001839 CrossRefPubMedPubMedCentralGoogle Scholar
  30. Laitinen L (1987) Griffonia simplicifolia lectins bind specifically to endothelial cells and some epithelial cells in mouse tissues. Histochem J 19:225–234CrossRefGoogle Scholar
  31. Lampa SJ, Potluri S, Norton AS, Laskowski MB (2004) A morphological technique for exploring neuromuscular topography expressed in the mouse gluteus maximus muscle. J Neurosci Methods 138:51–56.  https://doi.org/10.1016/j.jneumeth.2004.03.012 CrossRefPubMedGoogle Scholar
  32. Lexell J (1997) Muscle capillarization: morphological and morphometrical analyses of biopsy samples. Muscle Nerv Suppl 5:S110–S112CrossRefGoogle Scholar
  33. Lillioja S, Young AA, Culter CL, Ivy JL, Abbott WG, Zawadzki JK, Yki-Järvinen H, Christin L, Secomb TW, Bogardus C (1987) Skeletal muscle capillary density and fiber type are possible determinants of in vivo insulin resistance in man. J Clin Invest 80:415–424.  https://doi.org/10.1172/JCI113088 CrossRefPubMedPubMedCentralGoogle Scholar
  34. Lucas CA, Kang LHD, Hoh JFY (2000) Monospecific antibodies against the three mammalian fast limb myosin heavy chains. Biochem Biophys Res Commun 272:303–308.  https://doi.org/10.1006/bbrc.2000.2768 CrossRefPubMedGoogle Scholar
  35. Mårin P, Andersson B, Krotkiewski M, Björntorp P (1994) Muscle fiber composition and capillary density in women and men with NIDDM. Diabetes Care 17:382–386.  https://doi.org/10.2337/diacare.17.5.382 CrossRefPubMedGoogle Scholar
  36. Montero D (2016) Comment on Prior et al. Increased skeletal muscle capillarization independently enhances insulin sensitivity in older adults after exercise training and detraining. Diabetes 2015;64:3386-3395. Diabetes 65:e11–e12.  https://doi.org/10.2337/db15-1461 CrossRefPubMedGoogle Scholar
  37. Montero D, Oberholzer L, Haider T, Breenfeldt-Andersen A, Dandanell S, Meinild-Lundby AK, Maconochie H, Lundby C (2018) Increased capillary density in skeletal muscle is not associated with impaired insulin sensitivity induced by bed rest in healthy young men. Appl Physiol Nutr Metab 43:1334–1340.  https://doi.org/10.1139/apnm-2018-0195 CrossRefPubMedGoogle Scholar
  38. Nyholm B, Qu Z, Kaal A, Pedersen SB, Gravholt CH, Andersen JL, Saltin B, Schmitz O (1997) Evidence of an increased number of type IIb muscle fibers in insulin-resistant first-degree relatives of patients with NIDDM. Diabetes 46:1822–1828.  https://doi.org/10.2337/diab.46.11.1822 CrossRefPubMedGoogle Scholar
  39. Oana F, Takeda H, Hayakawa K, Matsuzawa A, Akahane S, Isaji M, Akahane M (2005) Physiological difference between obese (fa/fa) Zucker rats and lean Zucker rats concerning adiponectin. Metabolism 54:995–1001.  https://doi.org/10.1016/j.metabol.2005.02.016 CrossRefPubMedGoogle Scholar
  40. Prior SJ, Goldberg AP, Ortmeyer HK, Chin ER, Chen D, Blumenthal JB, Ryan AS (2015) Increased akeletal nuscle capillarization independently enhances insulin sensitivity in older adults after exercise training and detraining. Diabetes 64:3386–3395.  https://doi.org/10.2337/db14-1771 CrossRefPubMedPubMedCentralGoogle Scholar
  41. Saltin B, Lindgarde F, Houston M, Hörlin R, Nygaard E, Gad P (1979) Physical training and glucose tolerance in middle-aged men with chemical diabetes. Diabetes 28:30–32.  https://doi.org/10.2337/diab.28.1.s30 CrossRefPubMedGoogle Scholar
  42. Schaad L, Hlushchuk R, Barré S, Gianni-Barrera R, Haberthür D, Banfi A, Djonov V (2017) Correlative imaging of the murine hind limb vasculature and muscle tissue by microCT and light microscopy. Sci Rep 7:41842.  https://doi.org/10.1038/srep41842 CrossRefPubMedPubMedCentralGoogle Scholar
  43. Schiaffino S, Gorza L, Sartore S, Saggin L, Ausoni S, Vianello M, Gundersen K, Lømo T (1989) Three myosin heavy chain isoforms in type 2 skeletal muscle fibres. J Muscle Res Cell Motil 10:197–205CrossRefGoogle Scholar
  44. Solomon TPJ, Haus JM, Li Y, Kirwan JP (2011) Progressive hyperglycemia across the glucose tolerance continuum in older obese adults is related to skeletal muscle capillarization and nitric oxide bioavailability. J Clin Endocrinol Metab 96:1377–1384.  https://doi.org/10.1210/jc.2010-2069 CrossRefPubMedPubMedCentralGoogle Scholar
  45. Stuart CA, McCurry MP, Marino A, South MA, Howell MEA, Layne AS, Ramsey MW, Stone MH (2013) Slow-twitch fiber proportion in skeletal muscle correlates with insulin responsiveness. J Clin Endocrinol Metab 98:2027–2036.  https://doi.org/10.1210/jc.2012-3876 CrossRefPubMedPubMedCentralGoogle Scholar
  46. Torgan CE, Brozinick JT, Kastello GM, Ivy JL (1989) Muscle morphological and biochemical adaptations to training in obese Zucker rats. J Appl Physiol 67:1807–1813.  https://doi.org/10.1152/jappl.1989.67.5.1807 CrossRefPubMedGoogle Scholar
  47. Van Der Laarse WJ, Des Tombe AL, Lee-De Groot MBE, Diegenbach PC (1998) Size principle of striated muscle cells. Neth J Zool 48:213–223.  https://doi.org/10.1163/156854298x00075 CrossRefGoogle Scholar
  48. Van Wessel T, De Haan A, Van Der Laarse WJ, Jaspers RT (2010) The muscle fiber type-fiber size paradox: hypertrophy or oxidative metabolism? Eur J Appl Physiol 110:665–694.  https://doi.org/10.1007/s00421-010-1545-0 CrossRefPubMedPubMedCentralGoogle Scholar
  49. Vincent L, Féasson L, Oyono-Enguéllé S, Banimbek V, Denis C, Guarneri C, Aufradet E, Monchanin G, Martin C, Gozal D, Dohbobga M, Wouassi D, Garet M, Thiriet P, Messonnier L (2010) Remodeling of skeletal muscle microvasculature in sickle cell trait and α-thalassemia. Am J Physiol Heart Circ Physiol 298:H375–H384.  https://doi.org/10.1152/ajpheart.00812.2009 CrossRefPubMedGoogle Scholar
  50. Vital P, Larrieta E, Hiriart M (2006) Sexual dimorphism in insulin sensitivity and susceptibility to develop diabetes in rats. J Endocrinol 190:425–432.  https://doi.org/10.1677/joe.1.06596 CrossRefPubMedGoogle Scholar
  51. WHO (2018) Obesity and overweight. https://www.who.int/en/news-room/fact-sheets/detail/obesity-and-overweight. Accessed 9 Aug 2019
  52. Williams BA (2010) Toward a potential paradigm shift for the clinical care of diabetic patients requiring perineural analgesia: strategies for using the diabetic rodent model. Reg Anesth Pain Med 35:329–332.  https://doi.org/10.1097/AAP.0b013e3181e82e0b CrossRefPubMedPubMedCentralGoogle Scholar
  53. Yokomizo H, Inoguchi T, Sonoda N, Sakaki Y, Maeda Y, Inoue T, Hirata E, Takei R, Ikeda N, Fujii M, Fukuda K, Sasaki H, Takayanagi R (2014) Maternal high-fat diet induces insulin resistance and deterioration of pancreatic β-cell function in adult offspring with sex differences in mice. Am J Physiol Endocrinol Metab 306:E1163–E1175.  https://doi.org/10.1152/ajpendo.00688.2013 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of Medicine, Institute of AnatomyUniversity of LjubljanaLjubljanaSlovenia
  2. 2.Department of Animal Science, Chair for Genetics, Biotechnology and Immunology, Biotechnical FacultyUniversity of LjubljanaDomžaleSlovenia
  3. 3.Department of Physiology, Anthropology and Ethology, Biotechnical FacultyUniversity of LjubljanaLjubljanaSlovenia
  4. 4.Department of Biomathematics, Institute of PhysiologyCzech Academy of SciencePrague 4Czech Republic
  5. 5.Department of Anaesthesiology and Surgical Intensive TherapyUniversity Medical Centre LjubljanaLjubljanaSlovenia

Personalised recommendations