Application of the fractionator and vertical slices to estimate total capillary length in skeletal muscle

Artacho‐Pérula, Emilio; Roldán-Villalobos, R; Cruz‐Orive, Luis M.

doi:10.1046/j.1469-7580.1999.19530429.x

Cited by 9 publications

(5 citation statements)

References 33 publications

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“…A stereological method of vertical sections in combination with a fractionator design has been proposed (Artacho-Perula et al 1999) but, to our knowledge, this method has not been applied in skeletal muscle research.…”

Section: Introductionmentioning

confidence: 99%

Nerve injury affects the capillary supply in rat slow and fast muscles differently

et al. 2005

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The goal of this study was to determine the acute effects of permanent denervation on the length density of the capillary network in rat slow soleus (SOL) and fast extensor digitorum longus (EDL) muscles and the effect of short-lasting reinnervation in slow muscle only. Denervation was performed by cutting the sciatic nerve. Both muscles were excised 2 weeks later. Reinnervation was studied 4 weeks after nerve crush in SOL muscle only. Capillaries and muscle fibres were visualised by triple immunofluorescent staining with antibodies against CD31 and laminin and with fluorescein-labelled Griffonia (Bandeira) simplicifolia lectin. A recently developed stereological approach allowing the estimation of the length of capillaries adjacent to each individual fibre (Lcap/Lfib) was employed. Three-dimensional virtual test grids were applied to stacks of optical images captured with a confocal microscope and their intersections with capillaries and muscle fibres were counted. Interrelationships among capillaries and muscle fibres were demonstrated with maximum intensity projection of the acquired stacks of optical images. The course of capillaries in EDL seemed to be parallel to the fibre axes, whereas in SOL, their preferential direction deviated from the fibre axes and formed more cross-connections among neighbouring capillaries. Lcap/Lfib was clearly reduced in denervated SOL but remained unchanged in EDL, although the muscle fibres significantly atrophied in both muscle types. When soleus muscle was reinnervated, capillary length per unit fibre length was completely restored. The physiological background for the different responses of the capillary network in slow and fast muscle is discussed.

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Section: Introductionmentioning

confidence: 99%

Nerve injury affects the capillary supply in rat slow and fast muscles differently

et al. 2005

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“…Ongoing monitoring of the neovasculature is, therefore, important to ensure immediate vascularization and successful integration of the construct with surrounding tissue. Conventional assessment using histology and microscopy is limited in accuracy for a number of reasons, including availability of assays (5) and loss of three‐dimensional information (6). Quantitative assessment of angiogenesis is best made in vivo using a non‐invasive method for serial, three‐dimensional depiction of the vascular topology.…”

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confidence: 99%

Dynamic Gd‐DTPA enhanced MRI as a surrogate marker of angiogenesis in tissue‐engineered bladder constructs: A feasibility study in rabbits

Cheng

Chen

Babyn

et al. 2005

Magnetic Resonance Imaging

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Purpose: To evaluate the potential of dynamic contrastenhanced magnetic resonance imaging (DCE-MRI) to assess angiogenesis in tissue-engineered bladder constructs in a blinded animal study, and compare different analysis approaches and their correlation with microvessel density (MVD). Materials and Methods:Constructs fortified with vascular endothelial growth factor (VEGF) for enhanced vascularity were grafted onto the bladder in nine rabbits. DCE-MRI of Gd-DTPA uptake was performed and analyzed using Tofts' model, the area under the concentration time curve (AUC), and the uptake slope. DCE-MRI parameters were compared to MVD determined with CD31 immunohistochemistry.Results: Significantly increased MVD was found in the high VEGF group (20 ng/g of tissue) but not at low VEGF (10 ng/g) (2.3ϫ increase, P ϭ 0.035 vs. 1.1ϫ over control). Enhanced permeability at low VEGF was suggested by elevated K trans , but overall correlation to MVD was poor. Significant correlation to MVD was obtained with AUC 8min (r ϭ 0.705, P ϭ 0.034). Furthermore, AUC 8min provided the most precise discrimination between different VEGF preparations and was the only parameter to show a significant increase (P ϭ 0.0058) consistent with MVD changes at high VEGF. Conclusion:Findings support DCE-MRI for evaluating angiogenesis in bladder constructs and suggest vessel changes other than density. Future studies should incorporate larger contrast agents and permeability assessment to devise an optimal DCE-MRI strategy.

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“…Cruz-Orive and Howard CV developed the design-based formulae for total curve length in 1991 [71]. Vertical slice techniques have been used to estimate the length of blood vessels [72][73][74], dendritic trees [75], and, more recently, nephron tubule segments [76]. To treat the rat testicular albuginea as a thick section, we first describe the technique used to separate it from the testicular tissue and spread it in a plane.…”

Section: Stereological Estimate Of Length Of Cholinergic Fibersmentioning

confidence: 99%

Length measurement of cholinergic innervation in the tunica albuginea of rat testis

Santamaría,

Ingelmo

2023

Preprint

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The testicular albuginea is not a simple receptacle that contains the testis parenchyma. It is a biologically active structure involved, among other things, in the progression of sperm from the seminiferous tubules to the rete testis. This function, attributed to the albuginea myofibroblasts' contractile properties, is mediated by its vegetative innervation. Therefore, the quantification of innervation seems relevant to deepen the knowledge of the testicular envelope's physiology. The albuginea contains cholinergic nerve fibers that are not very evident and with a marked directional arrangement (anisotropy). These structural characteristics require that the quantification of the fibers' length be carried out using stereological measurements, which ensure the absence of bias in the process. The authors have developed stereological tools usually carried out on vertical VUR (vertical uniform random) sections in the present work. This approach's novelty is that the albuginea is treated "in toto" as if it were a thick section (slice) on which a cycloid grid is applied through different optical planes. Thus, on a pilot study of three specimens, cholinergic nerve fibers' length density was estimated, as well as the estimates of the coefficient of error of measurements and the biological variability of the sample. Furthermore, after estimating the albuginea volume, the mean of the fibers' absolute length can be obtained. This stereological approach can be used after visualization of the immunostained nerve fibers for various markers, as long as the reagents' penetration into the tissue is ensured. Moreover, it can be used in other biological structures, such as the intestine of small animals, which can also be processed as whole mounts and treated as thick VUR sections.

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Application of the fractionator and vertical slices to estimate total capillary length in skeletal muscle

Cited by 9 publications

References 33 publications

Nerve injury affects the capillary supply in rat slow and fast muscles differently

Nerve injury affects the capillary supply in rat slow and fast muscles differently

Dynamic Gd‐DTPA enhanced MRI as a surrogate marker of angiogenesis in tissue‐engineered bladder constructs: A feasibility study in rabbits

Length measurement of cholinergic innervation in the tunica albuginea of rat testis

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