The aim of this study was to determine whether capillarity in the denervated and reinnervated rat extensor digitorum longus muscle (EDL) is scaled by muscle fiber oxidative potential. We visualized capillaries adjacent to a metabolically defined fiber type and estimated capillarity of fibers with very high oxidative potential (O) vs fibers with very low oxidative potential (G). Capillaries and muscle fiber types were shown by a combined triple immunofluorescent technique and the histochemical method for NADH-tetrazolium reductase. Stacks of images were captured by a confocal microscope. Applying the Ellipse program, fibers were outlined, and the diameter, perimeter, cross-sectional area, length, surface area, and volume within the stack were calculated for both fiber types. Using the Tracer plug-in module, capillaries were traced within the three-dimensional (3D) volume, the length of capillaries adjacent to individual muscle fibers was measured, and the capillary length per fiber length (Lcap/Lfib), surface area (Lcap/Sfib), and volume (Lcap/Vfib) were calculated. Furthermore, capillaries and fibers of both types were visualized in 3D. In all experimental groups, O and G fibers significantly differed in girth, Lcap/Sfib, and Lcap/Vfib, but not in Lcap/Lfib. We conclude that capillarity in the EDL is scaled by muscle fiber size and not by muscle fiber oxidative potential.

Department of Biomathematics Institute of Physiology Academy of Sciences of the Czech Republic Prague Czech Republic

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Adolfson J, Ljungquist A, Tornling G, Unge G (1981) Capillary increase in the skeletal muscle of trained young and adult rats. J Physiol 310:529–532 PubMed PMC

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 82:231–234 PubMed

Andersen P, Henricksson J (1977) Capillary supply of the quadriceps femoris muscle of man: adaptive response to exercise. J Physiol 270:677–690 PubMed PMC

Badr I, Brown MD, Egginton S, Hudlicka O, Milkiewicz M, Verhaeg J (2003) Differences in local environment determine the site of physiological angiogenesis in rat skeletal muscle. Exp Physiol 88:565–568 PubMed

Brodal P, Ingjer F, Hermansen L (1977) Capillary supply of skeletal muscles fibers in untrained and endurance trained men. Am J Physiol 232:H7705–7712 PubMed

Brown MD, Cotter MA, Hudlicka O, Vrbova G (1976) The effects of different patterns of muscle activity on capillary density, mechanical properties and structure of slow and fast rabbit muscles. Pflugers Arch 361:241–250 PubMed

Čapek M, Brůža P, Janáček J, Karen P, Kubinová L, Vagnerová R (2009) Volume reconstruction of large tissue specimens from serial physical sections using confocal microscopy and correction of cutting deformations by elastic registration. Microsc Res Tech 72:110–119 PubMed

Capo LA, Sillau AH (1983) The effect of hypothyroidism on capillarity and oxidative capacity in rat soleus and gastrocnemius muscles. J Physiol 342:1–14 PubMed PMC

Čebašek V, Kubínová L, Janáček J, Ribarič S, Eržen I (2007) Adaptation of muscle fiber types and capillary network to acute denervation and shortlasting reinnervation. Cell Tissue Res 330:279–289 PubMed

Čebašek V, Kubínová L, Ribarič S, Eržen I (2004) A novel staining method for quantification and 3D visualisation of capillaries and muscle fibres. Eur J Histochem 48:151–158 PubMed

Čebašek V, Kubínová L, Ribarič S, Eržen I (2005) Capillary network in slow and fast muscles and in oxidative and glycolytic muscle fibres. Image Anal Stereol 24:51–58

Čebašek V, Radochová B, Kubínová L, Ribarič S, Eržen I (2006) Nerve injury affects the capillary supply in rat slow and fast muscles differently. Cell Tissue Res 323:305–312 PubMed

Charifi N, Kadi F, Feasson L, Costes F, Geyssant A, Denis C (2003) Enhancement of microvessel tortuosity in the vastus lateralis muscle of old men in response to endurance training. J Physiol 554:559–569 PubMed PMC

Cotter MA, Hudlicka O, Pette D, Staudte H, Vrbova H (1973) Changes of capillary density and enzyme pattern in fast rabbit muscles during long-term stimulation. J Physiol 230:34P–35P PubMed

Degens H, Deveci D, van Bemden AB, Hoofd LJC, Eginton S (2006) Maintenance of heterogeneity of capillary spacing is essential for adequate oxygenation in the soleus muscle of the growing rat. Microcirculation 13:467–476 PubMed

Degens H, Turek Z, Hoofd LJC, van't Hof MA (1992) The relationship between capillarisation and fibre types during compensatory hypertrophy of the plantaris muscle in the rat. J Anat 180:455–463 PubMed PMC

Deveci D, Marshall JM, Egginton S (2001) Relationship between capillary angiogenesis, fibre type and fibre size in chronic systemic hypoxia. Am J Physiol Heart Circ Physiol 281:H241–255 PubMed

Egginton S (1990) Numerical and areal density estimates of fibre type composition in a skeletal muscle (extensor digitorum longus). J Anat 168:73–80 PubMed PMC

Egginton S (2002) Temperature and angiogenesis. The possible role of mechanical factors in capillary growth. Comp Biochem Physiol A Mol Integr Physiol 132:773–787 PubMed

Egginton S, Hudlicka O (2000) Selective long term electrical stimulation of fast glycolytic fibres increases capillary supply but oxidative enzyme activity in rat skeletal muscles. Exp Physiol 85:567–573 PubMed

Gray SD, Renkin EM (1978) Microvascular supply in relation to fibre metabolic type in mixed skeletal muscles of rabbits. Microvasc Res 16:406–425 PubMed

Gute D, Laughin MH, Amann JF (1994) Regional changes in capillary supply in skeletal muscle of interval-sprint and low-intensity, endurance-trained rats. Microcirculation 1:183–193 PubMed

Hepple RT, Mathieu-Costello O (2001) Estimating the size of the capillary to fiber interface in skeletal muscle: a comparison of methods. J Appl Physiol 91:2150–2156 PubMed

Hepple RT, Vogel JE (2004) Anatomic capillarization is maintained in relative excess of fiber oxidative capacity in some skeletal muscles of late middle-aged rats. J Appl Physiol 96:2257–2264 PubMed

Ingjer F (1979) Effect of endurance training on muscle fibre ATPase activity, capillary supply and mitochondria content in man. J Physiol 294:419–432 PubMed PMC

Janáček J, Saxl I, Mao XW, Kubínová L (2007) Measurement of capillary length from 3D confocal images using image analysis and stereology. Focus on Microscopy, Valencia, Spain, April 10–13, Program and Abstract Book, p. 71

Karen P, Jirkovská M, Tomori Z, Demjénová E, Janáček J, Kubínová L (2003) Three-dimensional computer reconstruction of large tissue volumes based on composing series of high-resolution confocal images by GlueMRC and LinkMRC software. Microsc Res Tech 62:415–422 PubMed

Kubínová L, Janáček J, Ribarič S, Čebašek V, Eržen I (2001) Three-dimensional study of the capillary supply of skeletal muscle fibres using confocal microscopy. J Muscle Res Cell Motil 22:217–227 PubMed

Large J, Tyler KR (1985) Changes in capillary distribution in rat fast muscles following nerve crush and reinnervation. J Physiol 362:13–21 PubMed PMC

Mai JV, Edgerton VR, Barnard RJ (1970) Capillarity of red, white and intermediate muscle fibers in trained and untrained guinea pigs. Experientia 26:1222–1223 PubMed

Mathieu O, Cruz-Orive LM, Hoppeler H, Weibel ER (1983) Estimating length density and quantifying anisotropy in skeletal muscle capillaries. J Microsc 131:131–146 PubMed

Maxwell LC, White TP, Faulkner JA (1980) Oxidative capacity, blood flow, and capillarity of skeletal muscles. J Appl Physiol 49:627–633 PubMed

Novikoff AB, Shin WY, Drucker J (1961) Mitochondrial localization of oxidative enzymes: staining results with two tetrazolium salts. J Biophys Biochem Cytol 9:47–61 PubMed PMC

Panisello P, Torell JR, Esteva S, Pages T, Viscor G (2008) Capillary supply, fibre types and fibre morphometry in rat tibialis anterior and diaphragm muscle after intermittent exposure to hypobaric hypoxia. Eur J Appl Physiol 103:203–213 PubMed

Plyley MJ, Groom AC (1975) Geometrical distribution of capillaries in mammalian striated muscle. Am J Physiol 228:1376–1383 PubMed

Poole DC, Mathieu-Costello O, West JB (1989) Capillary tortuosity in rat soleus muscle is not affected by endurance training. Am J Physiol 256:H1110–1116 PubMed

Ribarič S, Stefanovska A, Brzin M, Kogovšek M, Krošelj P (1991) Biochemical, morphological, and functional changes during peripheral nerve regeneration. Mol Chem Neuropathol 15:143–157 PubMed

Romanul FCA (1965) Capillary supply and metabolism of muscle fibers. Arch Neurol 12:497–509 PubMed

Romanul FCA, Pollock M (1969) The parallelism of changes in oxidative metabolism and capillary supply of skeletal muscle fibers. In Locke S, ed. Modern Neurology. Boston, Little Brown, 203–213

Sillau AH (1985) Capillarity, oxidative capacity and fibre composition of the soleus and gastrocnemius muscles of rats in hypothyroidism. J Physiol 361:281–295 PubMed PMC

Sillau AH, Aquin L, Lechner A, Bui MV, Banchero N (1980) Increased capillary supply in skeletal muscle of guinea pigs acclimated to cold. Respir Physiol 42:233–245 PubMed

Škorjanc D, Jaschinsky F, Heine G, Pette D (1998) Sequential increases in capillarization and mitochondrial enzymes in low-frequency-stimulated rabbit muscle. Am J Physiol 274:C810–818 PubMed

Stål P, Eriksson P-O, Thornell LE (1996) Differences in capillary supply between human oro-facial, masticatory and limb muscles. J Muscle Res Cell Motil 17:183–197 PubMed

Sullivan SM, Pitman RN (1984) In vitro O2 uptake and histochemical fiber type of resting hamster muscles. J Appl Physiol 57:246–253 PubMed

Ziada AMA, Hudlicka O, Tyler KR, Wright AJA (1984) The effect of long-term vasodilatation on capillary growth and performance in rabbit heart and skeletal muscle. Cardiovasc Res 18:724–732 PubMed

Fiber-type composition and 3D capillary analysis of the human splenius capitis muscle

Skeletal muscle myosin heavy chain expression and 3D capillary network changes in streptozotocin-induced diabetic female mice

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

Capillary Network Morphometry of Pig Soleus Muscle Significantly Changes in 24 Hours After Death