Two new reliable immunohistochemical methods for simultaneous identification of capillaries, the three types of fibers and basal lamina in human skeletal muscle

Bailly, Mélina; Féasson, Léonard; Pereira, Bruno; Boileau, Amandine; Hourdé, Christophe; Germain, Natacha; Galusca, Bogdan; Courteix, Daniel; Thivel, David; Verney, Julien

doi:10.1007/s00418-020-01895-5

Cited by 5 publications

(8 citation statements)

References 51 publications

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“…In a recent publication, Al-Shammari et al (2019) presented a method to stain for types I, IIA, and IIX muscle fibers and the basal lamina using specific antibodies, while using the Ulex europaeus agglutinin (lectin; UEA I) I to label capillary endothelial cells in the same tissue section. Bailly et al (2020) have now presented a similar methodology, however replacing the UEA I lectin methodology with the endothelial cellspecific CD31 antibody. They tested five different protocols with various antibody cocktails and single-and two-step procedures to establish a time-and tissue-saving reliable immunohistochemical staining method for the three muscle fiber subtypes, the basal lamina (for muscle fiber morphometry measurements), and endothelial cells on a single tissue section (Fig.…”

Section: New Tissue Section Methods To Identify Muscle Fiber Subtypesmentioning

confidence: 99%

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In focus in HCB

Taatjes

Roth

2020

Histochem Cell Biol

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Section: New Tissue Section Methods To Identify Muscle Fiber Subtypesmentioning

confidence: 99%

“…4 Immunohistochemical staining of human skeletal muscle using method 1 (a) and method 3 (c) to reveal type I (blue), type IIA (green), type IIX (black) fibers, laminin (white), and endothelial cells (red). From Bailly et al (2020)…”

Section: New Tissue Section Methods To Identify Muscle Fiber Subtypesmentioning

confidence: 99%

In focus in HCB

Taatjes

Roth

2020

Histochem Cell Biol

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“…performed in two-steps to avoid cross-reactivity between BF-35 and anti-CD31 antibodies. This staining method has been previously described and validated (Bailly et al, 2020a). A slide scanner (Axio Scann.Z1, Carl Zeiss, Munich, Germany) was used for acquisition and ImageJ software (NIH, Bethesda, MD, United States) for image processing and analyses, as previously described (Bailly et al, 2020b).…”

Section: Immunofluorescencementioning

confidence: 99%

New Insights on Bone Tissue and Structural Muscle-Bone Unit in Constitutional Thinness

et al. 2022

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While few studies pointed out low bone mineral densities in constitutionally thin women, little is known about potential explanations. The objective was to further explore bone architecture in both women and men with constitutional thinness to investigate their mechanical muscle-bone coupling (or uncoupling). Thirty constitutionally thin people and 31 normal weight controls participated in the study. Body composition, hip structural analysis, and trabecular bone score were assessed by dual-energy X-ray absorptiometry, bone architecture using high-resolution peripheral quantitative computed tomography, and muscle explorations through histological staining on muscle biopsies. Thirty-two out of the 48 indexes relative to density, geometry, texture, and architecture of bones were found significantly lower (p < 0.05) in constitutionally thin individuals compared with controls. This observation was particularly pronounced in constitutionally thin men. Bone microarchitecture was more altered in weight-supporting bone (tibia) than in non-weight-supporting (radius) bone, which might refer to a normal physiological adaptation (Frost’s mechanostat theory). Yet, the heat-maps of correlations analyses showed many alterations of body weight or muscle associations with bone parameters in constitutionally thin individuals contrary to controls. Present results might support the idea of intrinsic disturbances of bone cells independently to the small muscle structure, particularly in men.

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“…These cells can be precisely identified with different well-characterised IHC and IF techniques [295]. Among them, and in the context of dystrophic muscles, the most used are biotinylate or fluorescein-labelled lectins such as Ulex europaeus agglutinin 1 (UEA 1, only for humans) [296] or isolectin B4 (IB4) from Griffonia simplicifolia [297], antibodies to the cluster of differentiation 31 (CD31) [12,298] for endothelial cells and to PDGFR-β, to neuron-glia antigen 2 (NG2) or to α-smooth muscle actin (α-SMA), for pericytes [12]. CD31, also recognised as platelet-endothelial cell adhesion molecule-1, is an adhesion molecule specifically expressed on endothelial cells, platelets and several immune cells [299].…”

Section: Changes In Capillarisationmentioning

confidence: 99%

“…Currently, by applying IF techniques, several groups have successfully identified different fibre types simultaneously with capillaries and laminin. For specific protocols, see: [140,296,298].…”

Section: Changes In Capillarisationmentioning

confidence: 99%

Histological Methods to Assess Skeletal Muscle Degeneration and Regeneration in Duchenne Muscular Dystrophy

Dubuisson

Versele

Planchon

et al. 2022

IJMS

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Duchenne muscular dystrophy (DMD) is a progressive disease caused by the loss of function of the protein dystrophin. This protein contributes to the stabilisation of striated cells during contraction, as it anchors the cytoskeleton with components of the extracellular matrix through the dystrophin-associated protein complex (DAPC). Moreover, absence of the functional protein affects the expression and function of proteins within the DAPC, leading to molecular events responsible for myofibre damage, muscle weakening, disability and, eventually, premature death. Presently, there is no cure for DMD, but different treatments help manage some of the symptoms. Advances in genetic and exon-skipping therapies are the most promising intervention, the safety and efficiency of which are tested in animal models. In addition to in vivo functional tests, ex vivo molecular evaluation aids assess to what extent the therapy has contributed to the regenerative process. In this regard, the later advances in microscopy and image acquisition systems and the current expansion of antibodies for immunohistological evaluation together with the development of different spectrum fluorescent dyes have made histology a crucial tool. Nevertheless, the complexity of the molecular events that take place in dystrophic muscles, together with the rise of a multitude of markers for each of the phases of the process, makes the histological assessment a challenging task. Therefore, here, we summarise and explain the rationale behind different histological techniques used in the literature to assess degeneration and regeneration in the field of dystrophinopathies, focusing especially on those related to DMD.

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Two new reliable immunohistochemical methods for simultaneous identification of capillaries, the three types of fibers and basal lamina in human skeletal muscle

Cited by 5 publications

References 51 publications

In focus in HCB

In focus in HCB

New Insights on Bone Tissue and Structural Muscle-Bone Unit in Constitutional Thinness

Histological Methods to Assess Skeletal Muscle Degeneration and Regeneration in Duchenne Muscular Dystrophy

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