Uranyl-Free Staining as a Suitable Contrasting Technique for Nuclear Structures at Transmission Electron Microscopy

Lacavalla, Maria Assunta; Cisterna, Barbara

doi:10.1007/978-1-0716-2675-7_18

Cited by 3 publications

(1 citation statement)

References 13 publications

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“…As control, some grids were floated on the incubation mixture without the primary antibody, and then treated as above. Sections were stained for 30 min at room temperature with Uranyl Less EM stain (Electron Microscopy Sciences, Hatfield, PA, USA), followed by Reynolds' lead citrate for 1 min (Lacavalla & Cisterna, 2023). Sections were then observed with a Philips Morgagni transmission electron microscope operating at 80 kV.…”

Section: Methodsmentioning

confidence: 99%

Physical training promotes remodeling of the skeletal muscle extracellular matrix: An ultrastructural study in a murine model of Down syndrome

Cisterna

Boschi

Lacavalla

et al. 2023

Microscopy Res & Technique

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Down syndrome (DS) is a genetically based disease caused by triplication of chromosome 21. DS is characterized by multi‐systemic premature aging associated with deficit in motor coordination, balance, and postural control. Using a morphological, morphometrical, and immunocytochemical ultrastructural approach, this study investigated in vastus lateralis muscle of Ts65Dn mouse, a murine model of DS, the effect of an adapted physical training on the extracellular matrix (ECM) characteristics and whether the forecasted exercise‐induced ECM remodeling impacts on sarcomere organization. Morphometry demonstrated thicker basement membrane and larger collagen bundles with larger interfibrillar spacing as well as irregularly arrayed myofibrils and lower telethonin density on Z‐lines in trisomic versus euploid sedentary mice. In agreement with the multi‐systemic premature aging described in DS, these ECM alterations were similar to those previously observed in skeletal muscle of aged mice. Adapted physical training induced remodeling of ECM in both trisomic and euploid mice, that is, enlargement of the collagen bundles associated with hypertrophy of collagen fibrils and reduction of the interfibrillar spacing. A re‐alignment of the myofibrils and a higher telethonin density on Z‐line was found in trisomic mice. Altogether, our findings suggest that physical training is an effective tool in limiting/counteracting the trisomy‐associated musculoskeletal structural anomalies. The current findings constitute a solid experimental background for further study investigating the possible positive effect of physical training on skeletal muscle performance.Research Highlights Vastus lateralis muscle of trisomic mice shows aging‐like alterations of extracellular matrix. Training promotes extracellular matrix remodeling. Training may be an effective tool to counteract trisomy‐associated alterations of skeletal muscle.

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Section: Methodsmentioning

confidence: 99%

Physical training promotes remodeling of the skeletal muscle extracellular matrix: An ultrastructural study in a murine model of Down syndrome

Cisterna

Boschi

Lacavalla

et al. 2023

Microscopy Res & Technique

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Aged gastrocnemius muscle of mice positively responds to a late onset adapted physical training

Cisterna,

Lofaro,

Lacavalla

et al. 2023

Front. Cell Dev. Biol.

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Introduction: A regular physical training is known to contribute to preserve muscle mass and strength, maintaining structure and function of neural and vascular compartments and preventing muscle insulin resistance and inflammation. However, physical activity is progressively reduced during aging causing mobility limitations and poor quality of life. Although physical exercise for rehabilitation purposes (e.g., after fractures or cardiovascular events) or simply aiming to counteract the development of sarcopenia is frequently advised by physicians, nevertheless few data are available on the targets and the global effects on the muscle organ of adapted exercise especially if started at old age.Methods: To contribute answering this question for medical translational purposes, the proteomic profile of the gastrocnemius muscle was analyzed in 24-month-old mice undergoing adapted physical training on a treadmill for 12 weeks or kept under a sedentary lifestyle condition. Proteomic data were implemented by morphological and morphometrical ultrastructural evaluations.Results and Discussion: Data demonstrate that muscles can respond to adapted physical training started at old age, positively modulating their morphology and the proteomic profile fostering protective and saving mechanisms either involving the extracellular compartment as well as muscle cell components and pathways (i.e., mitochondrial processes, cytoplasmic translation pathways, chaperone-dependent protein refolding, regulation of skeletal muscle contraction). Therefore, this study provides important insights on the targets of adapted physical training, which can be regarded as suitable benchmarks for future in vivo studies further exploring the effects of this type of physical activity by functional/metabolic approaches.

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Physical Training Chronically Stimulates the Motor Neuron Cell Nucleus in the Ts65Dn Mouse, a Model of Down Syndrome

Inguscio

Lacavalla

Cisterna

et al. 2023

Cells

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Down syndrome (DS) is a genetically-based disease based on the trisomy of chromosome 21 (Hsa21). DS is characterized by intellectual disability in association with several pathological traits among which early aging and altered motor coordination are prominent. Physical training or passive exercise were found to be useful in counteracting motor impairment in DS subjects. In this study we used the Ts65Dn mouse, a widely accepted animal model of DS, to investigate the ultrastructural architecture of the medullary motor neuron cell nucleus taken as marker of the cell functional state. Using transmission electron microscopy, ultrastructural morphometry, and immunocytochemistry we carried out a detailed investigation of possible trisomy-related alteration(s) of nuclear constituents, which are known to vary their amount and distribution as a function of nuclear activity, as well as the effect of adapted physical training upon them. Results demonstrated that trisomy per se affects nuclear constituents to a limited extent; however, adapted physical training is able to chronically stimulate pre-mRNA transcription and processing activity in motor neuron nuclei of trisomic mice, although to a lesser extent than in their euploid mates. These findings are a step towards understanding the mechanisms underlying the positive effect of physical activity in DS.

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Uranyl-Free Staining as a Suitable Contrasting Technique for Nuclear Structures at Transmission Electron Microscopy

Cited by 3 publications

References 13 publications

Physical training promotes remodeling of the skeletal muscle extracellular matrix: An ultrastructural study in a murine model of Down syndrome

Physical training promotes remodeling of the skeletal muscle extracellular matrix: An ultrastructural study in a murine model of Down syndrome

Aged gastrocnemius muscle of mice positively responds to a late onset adapted physical training

Physical Training Chronically Stimulates the Motor Neuron Cell Nucleus in the Ts65Dn Mouse, a Model of Down Syndrome

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