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المصدر: Acta Histochemica. 98:345-355
مصطلحات موضوعية: Male, musculoskeletal diseases, congenital, hereditary, and neonatal diseases and abnormalities, Histology, Biology, Models, Biological, Sarcolemma, Sarcoglycans, Diaphorase, medicine, Animals, Rats, Wistar, Fluorescent Antibody Technique, Indirect, Muscle, Skeletal, chemistry.chemical_classification, Membrane Glycoproteins, Skeletal muscle, Cell Biology, General Medicine, musculoskeletal system, Rats, Turtles, Cell biology, Isoenzymes, Nitric oxide synthase, Cytoskeletal Proteins, Sarcoglycan, Enzyme, medicine.anatomical_structure, chemistry, Biochemistry, biology.protein, Nitric Oxide Synthase, Glycoprotein, Dystrophin, Chickens
الوصف: Previous studies have shown the association of NOS I with the sarcolemma in mammalian striated muscle fibers, implicating the dystrophin complex (DC) as a major anchor for the enzyme. The potential role of the sarcoglycan subcomplex, especially of alpha-sarcoglycan (adhalin), as part of the DC in holding of NOS I in the sarcolemmal position was examined by carrying out a comparative study on the distribution of NOS I, dystrophin, dystrophin-associated glycoproteins (DAG) and alpha-sarcoglycan in various skeletal muscles of non-mammals. Rat muscles were included since they reflect the situation in mammals. Catalytic NOS-associated diaphorase (NOSaD) activity as well as NOS I and DAG immunoreactivities were positive in the saracolemma region of skeletal muscle fibers of rats, chicken, and turtles. Adhalin immunoreactivity was present in the rat but absent in the chicken and turtle muscle surface membrane. These data suggest that alpha-sarcoglycan and therefore the entire sarcoglycan subcomplex may not be needed for localizing NOS I to the sarcolemma in these non-mammalian species. This may hold for skeletal muscle fibers in general.
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المصدر: European Journal of Neuroscience. 8:202-210
مصطلحات موضوعية: Nervous system, medicine.medical_specialty, Sympathetic Nervous System, Population, Gene Expression, Cytochrome c Group, Axonal Transport, Polymerase Chain Reaction, Neuroprotection, Iodine Radioisotopes, Transforming Growth Factor beta, Internal medicine, medicine, Animals, Chromaffin Granules, RNA, Messenger, Rats, Wistar, education, Cells, Cultured, Neurons, education.field_of_study, biology, business.industry, General Neuroscience, NADPH Dehydrogenase, Spinal cord, Rats, medicine.anatomical_structure, Endocrinology, Spinal Cord, Adrenal Medulla, GDF7, Adrenal Cortex, Axoplasmic transport, biology.protein, Female, Fibroblast Growth Factor 2, business, Adrenal medulla, Neuroscience, Neurotrophin
الوصف: Transforming growth factors beta (TGF-beta), a family of pleiotropic cytokines, are widely distributed in the developing and adult nervous system. In order to further determine the neural functions of TGF-beta, we have localized the TGF-beta isoforms 1, 2 and 3 in the adult rat adrenal medulla and studied the neuroprotective capacity of one representative family member, TGF-beta 2, for those spinal cord neurons which innervate adrenal chromaffin cells and which die after destruction of the adrenal medulla. Unilateral electrothermal destruction of the adrenal medulla led to the disappearance of 25% of sympathetic preganglionic neurons, which are located in the intermediolateral (IML) column of thoracic spinal cord segments 7-10 and can be selectively marked by NADPH-diaphorase. The neurons which disappeared following adrenomedullectomy constitute the full set of neurons that innervate the adrenal medulla. Implantation of gelfoam soaked with 0.5 micrograms TGF-beta 2 into the adrenal wound cavity rescued all spinal cord neurons in the IML ipsilaterally to the lesioned side. Cytochrome c was not effective. Injections of [125I]TGF-beta 2 into the adrenal medulla did not result in retrograde transport and subsequent labelling of spinal cord neurons, suggesting that TGF-beta may exert its neuroprotective actions by indirect mechanisms. TGF-beta applied to cultured adrenocortical cells did not overtly increase the amount of mRNA for fibroblast growth factor-2, an established trophic molecule for sympathetic preganglionic spinal cord neurons. The mechanisms by which TGF-beta exerts its neurotrophic effect are therefore unclear. Even so, our data provide the first evidence that TGF-beta may play an important role in vivo in the control of maintenance of a population of spinal cord neurons.
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المؤلفون: Katsumasa Goto, Yoshitaka Ohno, Elisa Bianchini, Daniela Danieli-Betto, Antonio Musarò, Yoshinobu Ohira, Jean-François Desaphy, Giulia Maria Camerino, Anne Picard, Takashi Ohira, Michele Salanova, Stefano Ciciliot, Dorianna Sandonà, Gabriella Dobrowolny, Romeo Betto, Martina Gutsmann, Fuminori Kawano, Diana Conte, Dieter Blottner, Elena Germinario, Sandra Furlan, Stefano Schiaffino, Gudrun Schiffl, Naoya Nakai
المصدر: PLoS ONE
PLoS ONE, Vol 7, Iss 3, p e33232 (2012)
PLoS ONE; Vol 7مصطلحات موضوعية: muscle atrophy, Male, Anatomy and Physiology, fast and slow skeletal muscle fibers, lcsh:Medicine, Astronomical Sciences, Nitric Oxide Synthase Type I, E3 ubiquitin ligases, Biochemistry, law.invention, Mice, Potassium Channels, Calcium-Activated, 0302 clinical medicine, law, Myosin, Molecular Cell Biology, Insulin-Like Growth Factor I, lcsh:Science, Musculoskeletal System, 0303 health sciences, Multidisciplinary, Weightlessness, musculoskeletal, neural, and ocular physiology, Animal Models, Space Exploration, musculoskeletal system, ion-channels, Adaptation, Physiological, Immunohistochemistry, Potassium channel, Muscle atrophy, Up-Regulation, Slow-Twitch Muscle Fiber, medicine.anatomical_structure, skeletal muscle atrophy, MICROGRAVITY, gene expression, ubiquitin, Muscle, medicine.symptom, Cellular Types, tissues, signal transduction, Research Article, medicine.medical_specialty, Histology, Ubiquitin-Protein Ligases, Down-Regulation, microgravity, Biology, Spaceflight, 03 medical and health sciences, Model Organisms, Internal medicine, medicine, Animals, Muscle, Skeletal, 030304 developmental biology, Soleus muscle, Myosin Heavy Chains, Interleukin-6, lcsh:R, Skeletal muscle, Proteins, Space Flight, Rats, Mice, Inbred C57BL, Endocrinology, Metabolism, lcsh:Q, 030217 neurology & neurosurgery
الوصف: The effect of microgravity on skeletal muscles has so far been examined in rat and mice only after short-term (5-20 day) spaceflights. The mice drawer system (MDS) program, sponsored by Italian Space Agency, for the first time aimed to investigate the consequences of long-term (91 days) exposure to microgravity in mice within the International Space Station. Muscle atrophy was present indistinctly in all fiber types of the slow-twitch soleus muscle, but was only slightly greater than that observed after 20 days of spaceflight. Myosin heavy chain analysis indicated a concomitant slow-to-fast transition of soleus. In addition, spaceflight induced translocation of sarcolemmal nitric oxide synthase-1 (NOS1) into the cytosol in soleus but not in the fast-twitch extensor digitorum longus (EDL) muscle. Most of the sarcolemmal ion channel subunits were up-regulated, more in soleus than EDL, whereas Ca(2+)-activated K(+) channels were down-regulated, consistent with the phenotype transition. Gene expression of the atrophy-related ubiquitin-ligases was up-regulated in both spaceflown soleus and EDL muscles, whereas autophagy genes were in the control range. Muscle-specific IGF-1 and interleukin-6 were down-regulated in soleus but up-regulated in EDL. Also, various stress-related genes were up-regulated in spaceflown EDL, not in soleus. Altogether, these results suggest that EDL muscle may resist to microgravity-induced atrophy by activating compensatory and protective pathways. Our study shows the extended sensitivity of antigravity soleus muscle after prolonged exposition to microgravity, suggests possible mechanisms accounting for the resistance of EDL, and individuates some molecular targets for the development of countermeasures.
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المؤلفون: Salanova, M, Betto, R, Conte, D, Danieli, Daniela, Desaphy, J. F., Musarò, A, Ohira, Y, Sandona', Dorianna, Schiaffino, S, Dieter, Blottner
URL الوصول: https://explore.openaire.eu/search/publication?articleId=od______3657::199778c97a3ac1c2e38bc2275b3f26aa
http://hdl.handle.net/11577/2578459 -
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المؤلفون: Martina Gutsmann, Michele Salanova, Elena Bortoloso, Gudrun Schiffl, Dieter Blottner, Pompeo Volpe, Dieter Felsenberg, Sandra Furlan, Daniel L. Belavy
مصطلحات موضوعية: Male, medicine.medical_specialty, Neuromuscular Junction, Motor nerve, Biology, Biochemistry, Models, Biological, Vibration, Neuromuscular junction, Homer Scaffolding Proteins, Postsynaptic potential, Internal medicine, Genetics, medicine, Animals, Humans, RNA, Messenger, Rats, Wistar, Muscle, Skeletal, Molecular Biology, Transcription factor, Exercise, DNA Primers, Denervation, Messenger RNA, Base Sequence, NFATC Transcription Factors, Skeletal muscle, NFAT, Resistance Training, Adaptation, Physiological, Muscle Denervation, Muscular Disorders, Atrophic, Recombinant Proteins, Rats, Endocrinology, medicine.anatomical_structure, Gene Expression Regulation, Carrier Proteins, Bed Rest, Biotechnology, Signal Transduction
الوصف: Protein calcium sensors of the Homer family have been proposed to modulate the activity of various ion channels and nuclear factor of activated T cells (NFAT), the transcription factor modulating skeletal muscle differentiation. We monitored Homer expression and subcellular localization in human skeletal muscle biopsies following 60 d of bedrest [Second Berlin Bedrest Study (BBR2-2)]. Soleus (SOL) and vastus lateralis (VL) biopsies were taken at start (pre) and at end (end) of bedrest from healthy male volunteers of a control group without exercise (CTR; n=9), a resistive-only exercise group (RE; n=7), and a combined resistive/vibration exercise group (RVE; n=7). Confocal analysis showed Homer immunoreactivity at the postsynaptic microdomain of the neuromuscular junction (NMJ) at bedrest start. After bedrest, Homer immunoreactivity decreased (CTR), remained unchanged (RE), or increased (RVE) at the NMJ. Homer2 mRNA and protein were differently regulated in a muscle-specific way. Activated NFATc1 translocates from cytoplasm to nucleus; increased amounts of NFATc1-immunopositive slow-type myonuclei were found in RVE myofibers of both muscles. Pulldown assays identified NFATc1 and Homer as molecular partners in skeletal muscle. A direct motor nerve control of Homer2 was confirmed in rat NMJs by in vivo denervation. Homer2 is localized at the NMJ and is part of the calcineurin-NFATc1 signaling pathway. RVE has additional benefit over RE as countermeasure preventing disuse-induced neuromuscular maladaptation during bedrest.
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7276b0da08f37c6369d16b854108ba67
http://hdl.handle.net/11577/2488239 -
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المؤلفون: Martina Gutsmann, Daniel L. Belavy, Michele Salanova, Dieter Blottner, Gudrun Schiffl, Dieter Felsenberg
المصدر: The FASEB Journal. 24
مصطلحات موضوعية: Resistive touchscreen, medicine.anatomical_structure, Protein S-nitrosylation, medicine.medical_treatment, Genetics, medicine, Biophysics, Skeletal muscle, Biology, Bed rest, Molecular Biology, Biochemistry, Biotechnology
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المؤلفون: Wolfgang Ertel, Annekatrin Aue, Karym El-Sayed, Katharina Stoelzel, Gundula Schulze-Tanzil, Ulrike Marzahn, Dieter Blottner, A. Haisch, Benjamin Kohl, Thilo John, Maren Kuhne
المصدر: International Journal of Molecular Medicine. 25
مصطلحات موضوعية: Cartilage, Articular, Alginates, Swine, Cell Culture Techniques, Type II collagen, SOX9, Chondrocyte, Chondrocytes, Genetics, medicine, Articular cartilage repair, Animals, Humans, Regeneration, Collagen Type II, Cells, Cultured, Chemistry, Cartilage, General Medicine, Chondrogenesis, Coculture Techniques, Cell biology, Transplantation, medicine.anatomical_structure, Immunology, Ear Cartilage, Type I collagen
الوصف: Cartilage injury remains a challenge in orthopedic surgery as articular cartilage only has a limited capacity for intrinsic healing. Autologous chondrocyte transplantation (ACT) is a suitable technique for cartilage repair, but requires articular cartilage biopsies for autologous chondrocyte expansion. The use of heterotopic chondrocytes derived from non-articular cartilage sources such as auricular chondrocytes may be a novel approach for ACT. The aim of the study is to evaluate whether co-cultured articular/auricular chondrocytes exhibit characteristics comparable to articular chondrocytes. Analysis of the proliferation rate, extracellular cartilage matrix (ECM) gene and protein expression (type II and I collagen, elastin, lubricin), beta1-integrins and the chondrogenic transcription factor sox9 in articular/auricular chondrocytes was performed using RTD-PCR, flow cytometry, immunofluorescence microscopy and Western blot analysis. Additionally, three-dimensional (3D) chondrocyte mono- and co-cultures were established. The proliferative activity and elastin gene expression were lower and that of type II collagen and lubricin was higher in articular compared with auricular chondrocytes. The species generally did not influence the chondrocyte characteristics, with the exception of type I collagen and sox9 expression, which was higher in porcine but not in human articular chondrocytes compared with both types of auricular chondrocytes. beta1-integrin gene expression did not differ significantly between the chondrocyte types. The type II collagen gene and protein expression was higher in articular chondrocyte monocultures and was slightly higher in co-cultures compared with monocultured auricular chondrocytes. Both chondrocyte types survived in co-culture. Despite their differing expression profiles, co-cultures revealed some adjustment in the ECM expression of both chondrocyte types.
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0c1c658418c4921a342e3b6919a00568
https://doi.org/10.3892/ijmm_00000394 -
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المؤلفون: Peter Ueblacker, Lutz Hänsel, Gerhard Luttke, Peter Mundinger, Klaus Eder, William E. Garrett, Helmut Hoffmann, Andreas Schlumberger, Alfred Binder, Andreas Betthäuser, Norbert Maassen, Johannes Böck, Martin Flueck, Michael Hufnagl, Benedikt Schoser, Wilhelm Bloch, Dieter Blottner, Bernhard Brenner, Hans-Wilhelm Müller-Wohlfahrt, Jan Ekstrand
المصدر: Muskelverletzungen im Sport
مصطلحات موضوعية: business.industry, Medicine, business
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::1b5aef373c3632f13eb8110915f5ded2
https://doi.org/10.1055/b-002-43892 -
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المؤلفون: Dieter Blottner, H. G. Baumgarten
المصدر: The Journal of Comparative Neurology. 316:45-55
مصطلحات موضوعية: medicine.medical_specialty, Sympathetic Nervous System, Central nervous system, Amidines, Biology, Choline O-Acetyltransferase, Parasympathetic Nervous System, Internal medicine, Glial Fibrillary Acidic Protein, Neural Pathways, medicine, Animals, Cholinergic neuron, Neurons, Staining and Labeling, Histocytochemistry, General Neuroscience, Rats, Inbred Strains, Spinal cord, Immunohistochemistry, Molecular biology, Choline acetyltransferase, Rats, Nitric oxide synthase, Autonomic nervous system, medicine.anatomical_structure, Endocrinology, Spinal Cord, biology.protein, Cholinergic, Female, Amino Acid Oxidoreductases, Nitric Oxide Synthase, Immunostaining
الوصف: Nitric oxide synthetase (NOS) can be selectively stained in neurons by either NAJIPHdiaphorase (i.e., NOS)-histochemistry or immunohistochemistry with antibodies raised against NOS, which apparently label identical reactive sites (Hope, B.T., G.J. Michael, K.M. Knigge, and S.R. Vincent, Proc. Natl. Acad. Sci. USA 88:2811-2814, '91). We provide histochemical evidence for the existence of a neuron-specific NOS-activity in autonomic neurons of the thoracic spinal cord. Among the four main preganglionic cell clusters investigated at midthoracic levels, Th7-10, the intermediolateral (1ML)-cell column was the most prominently stained cell group. The histochemical staining was absent in other spinal cord neurons and non-neuronal cells, e.g., GFAP-positive glial cells. Staining was completely blocked by ""-nitroL-arginine (L-"A), a potent NOS-inhibitor for brain and peripheral autonomic neurons, but was still observed in the presence of another NOS-inhibitor, Nu-monomethyl-L-arginine (MeArg). The NOS-activity co-localized with nearly half of the ChAT-immunostained neurons located in the mid-thoracic IML-cell column as quantified by cell counts in single and double-stained tissue sections. We conclude that NOS-activity-containing neurons represent a distinct group among cholinergic IML-neurons, which suggests a more general function of this newly defined subpopulation of the spinal cord autonomic system. In vivo Fast blue retrograde labeling combined with histochemical staining and immunostaining revealed that sympathoadrenal projection neurons belong to the distinct NOS and ChAT-positive IML-cell group. The results are consistent with the idea of a crucial role of NOS and the related L-argininedependent nitric oxide biosynthesis for adrenomedullary innervation and autonomic control of adrenal gland function. In the adult rat, the intermediate region of the thoracic spinal cord (comprising laminae VI, VII, and X according to the classification of Rexed, '54) contains sympathetic preganglionic neurons (SPNs) that form distinct cholinergic cell clusters (Navaratnam and Lewis, '70; Borges and Iversen, '86) and are referred to as the central autonomic (CA)
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المؤلفون: Michele Salanova, Dieter Blottner, Gudrun Schiffl
المصدر: Histochemistry and cell biology. 132(4)
مصطلحات موضوعية: Gene isoform, medicine.medical_specialty, Histology, SERCA, medicine.medical_treatment, Bed rest, Protein expression, Quadriceps Muscle, Sarcoplasmic Reticulum Calcium-Transporting ATPases, Myofibrils, Internal medicine, medicine, Myocyte, Humans, Protein Isoforms, Molecular Biology, Exercise, Chemistry, Cell Biology, S-Nitrosylation, Muscular Disorders, Atrophic, Medical Laboratory Technology, Cytosol, Endocrinology, Muscle Fibers, Slow-Twitch, Lower Extremity, Female, Homeostasis, Bed Rest
الوصف: We monitored changes in SERCA isoform specific expression and S-nitrosylation in myofibers of lower limb soleus (SOL) and vastus lateralis (VL) muscle biopsies before and after 60 days of voluntary long term bed rest (BR) without (BR-CTRL group, n = 8) and with exercise countermeasure (BR-EX group, n = 8). Before BR, a typical myofiber type-specific distribution of fast and slow SERCA1/2a isoforms was seen. After BR, a subpopulation (approx. 15%) of slow myofibers in BR-CTRL additionally expressed the fast SERCA1a isoform which was not seen in BR-EX. After BR, SERCA1a S-nitrosylation patterns analyzed by the biotin-switch assay decreased in disused SOL only but increased in both muscles following exercise. Differential SERCA1a S-nitrosylation and SERCA1a/2a co-expression in subsets of slow myofibers should be considered as signs of an altered cytosolic Ca(2+) homeostasis following chronic muscle disuse. Exercise preserved myofiber type-specific SERCA1a expression and S-nitrosylation in VL and SOL in a different way, suggesting muscle-specific responses to the countermeasure protocol applied during bed rest.
