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Molecular Mechanisms in Muscle Development and Function
Molekularni mehanizmi razvoja in delovanja skeletne mišice
Start: 1/1/2004
End: 12/31/2008
Homepage: http://sicris.izum.si/search/prg.aspx?lang=slv&id=3680

Project Status History
unknown 1/1/2004
 
Keywords
skeletal muscle, mammals, plasticity, regeneration, ageing, myosin heavy chains, capillaries, satellite cells, myonuclei, neuromuscular junction, acetilcholinesterase mRNA, agrin,culture of human skeletal muscle, insulin resistence, RT-PCR, in situ hybrid
skeletna mišica, sesalci-, plastičnost-, regeneracija,-staranje,-težke verige miozina,-kapilare,-satelitne celice,-mišična jedra, živčnomišični stik, acetilholinesteraza, acetilholinestrazna mRNA, agrin, kultura človeške skeletne mišice, insulinska rezist

Abstract
The program joins two research groups: 'Skeletal muscle: its structure and function'and 'Human muscle under the in vitro conditions'.The program proposed by group A is dealing with the morphological, morphometric and immunohistochemical analysis of normal and diseased muscles. It consists of three research directions:I. Morphological and histochemical characteristics of skeletal muscles:We shall continue investigating the morphology and the expression pattern of myosin heavy chain isoforms (MyHC) and their transcripts in skeletal muscles of bigger mammals. Human extraocular muscles will be analysed in different age periods to prove that human EOM do not age. Special emphasis will be given to the capillary density per muscle fibre. II. Molecular mechanisms underlying the acute myopathic changes, accompanying critical conditions. Recent investigations revealed active role of the skeletal muscle fibre in the signalling processes, triggered by different types of cytokines under the stress conditions like septic shock. In our preliminary studies we demonstrated release of IL-6 from the cultured human skeletal muscle fibre after the treatments with TNF-a. These investigation will be continued in this part of the program.III. Molecular mechanisms responsible for the insulin resistance in the human skeletal muscle fibre. Insulin resistance is the major cause of hyperglycemia in type 2 diabetics and is therefore intensively investigated. The experimental model of the in vitro innervated human muscle allows studies of the molecular mechanisms underlying insulin-induced exocytosis, which is impaired in the insulin-resistant muscle fibres. In collaboration with the research group at the Institute of Pathophysiology, which is fully equipped to follow the processes of exocytosis, we shall investigate molecular mechanisms underlying the insulin-induced translocation of the glucose transporters 4 (GLUT4) bearing vesicles to theII. Plasticity of skeletal muscle in physiological, experimental and pathological conditionsPlasticity of skeletal muscles that has been thoroughly studied in our previous research will further be investigated either on experimental models, like heterochronous isotransplantation, heterotopic transplantation of the rat slow and fast muscles as well as axotonotomy and axonotmesis. Additionally, plasticity will be studied in human muscles in inactivity and in the critical illness myopathy. Special attention will be paid to the influence of the changed innervation, chorticosteroid and thyroid hormones on the expression of the myosin heavy chain isoforms and their transcripts as well as to the myogenic factors and to the changed capillary supply.A part of our research will be devoted to the investigation of the biophysical characteristics of normal and dystrophic muscles and to the morphometric analysis of the muscles in patients with the myotonic dystrophy III Epidemiology of the limb-girdle muscular dystrophyWe shall continue to study the epidemiology of the 'limb-girdle dystrophy' in Slovenia.The research program proposed by group B is focused on the molecular mechanisms underlying various aspects of the development and function of the human skeletal muscle fibre. In most studies we will employ the in vitro system in which human muscle fibre is innervated by the motor neuron extending from the explant of the embryonic rat spinal cord. The main research directions will be the following:I. Molecular mechanism regulating synaptogenesis and function of the neuromuscular junction (NMJ)By "gene silencing" we will investigate the roles and expression of specific proteins participating in the formation and maintenance of the NMJ. This part of our research will be focused on agrin, AChE and BuChE, which are all important, basal lamina bound constituents of the NMJ. This part of our program includes continuation of our previous research on the structure - function relationship of the AChE.
Program obsega dve programski podskupini:A. Struktura in funkcija skeletne mišiceB. Človeška mišica v razmerah in vitroV predlaganem programu skupine A želimo proučevati morfologijo ter morfometrične in imunohistokemične značilnosti skeletnih mišic. Program ima tri sklope:I. Morfometrične in histokemične značilnosti skeletnih mišicNadaljevali bomo z raziskavami morfoloških značilnosti in izražanja izooblik težkih verig miozina ter transkriptov zanje v mišicah velikih sesalcev. V ekstraokularnih mišicah človeka, ki jih bomo analizirali v različnih starostnih obdobjih, želimo dokazati, da se te mišice ne starajo.II. Plastičnost skeletne mišice v fizioloških, eksperimentalnih in patoloških pogojihŠtudije plastičnosti, ki smo jo podrobneje proučevali že v preteklem obdobju, nameravamo nadaljevati na eksperimentalnih modelih kot so heterokrona izotransplantacija, heterotopična transplantacija podganje hitre in počasne mišice, aksonotomija in aksonotmeza, pa tudi na humanih mišicah pri inaktivitetni atrofiji in pri akutni miopatiji kritično bolnega s selektivno izgubo miozinskih filamentov. Posebno pozornost bomo posvetili vplivu spremenjene inervacije, kortikosteroidnih in tireoidnih hormov na ekspresijo izooblik MyHC in transkriptov zanje, izražanju miogenih faktorjev in spremenjeni kapilarni mreži. Del raziskav je namenjen proučevanju biofizikalnih lastnosti normalnih in distrofičnih mišic ter morfometrični analizi mišic pri bolnikih z miotonično distrofijo.III. Epidemiologija ramensko-medenične mišične distrofijeZadnji sklop proučuje epidemiologijo ramensko-medeničnih oblik mišične distrofije v SlovenijiPredlagani program skupine B se loteva preučevanja molekularnih mehanizmov, ki uravnavajo delovanje skeletne mišice. Posebna pozornost je pri tem posvečena mehanizmom, ki sodelujejo pri oživčenju skeletnega mišičnega vlakna; ker pri tem procesu sodelujejo poleg skeletnih mišičnih vlaken tudi motonevroni, ki izvirajo iz hrbtenjače, so v ta del programa vključene tudi raziskave na ravni centralnega živčevja, predvsem na ravni same hrbtenjače. Program raziskav ima naslednje smeri: I. Molekularni mehanizmi, ki uravnavajo sinaptogenezo in delovanje živčnomišičnega stika (ŽMS) : S pomočjo tehnik "utišanja genov" bomo skušali ugotoviti pomen posameznih beljakovinskih molekul, ki se sintetizirajo v motonevronu in sodelujejo pri nastajanju ŽMS; drugi del raziskav bo posvečen mehanizmom, ki uravnavajo ekspresijo teh komponent ŽMS v motonevronu v posameznih fazah razvoja ŽMS. Pri tem bo večina raziskav posvečena molekulam kot so agrin, AChE in BuChE. V okviru tega sklopa bomo nadaljevali tudi raziskave na ravni ugotavljanja povezanosti med strukturo in funkcijo AChE. II. Molekularni mehanizmi akutnih miopatičnih sprememb, ki spremljajo nekatera kritična stanja, kot je npr septični šok: novejše raziskave so pokazale, da skeletno mišično vlakno aktivno sodeluje pri procesih, ki se prožijo prek različnih vrst citokinov, kar ima poseben pomen v raznih patoloških stanjih kot je n pr. septični šok. Preliminarne študije, ki smo jih opravili v preteklem obdobju na kulturi človeške mišice, so to potrdile. V okviru programa nameravamo zato nadaljevati z raziskavami molekularnih mehanizmov, predvsem tistih, prek katerih TNF-alfa vpliva na izločanje IL-6 iz človeške skeletne mišice v kulturi. III. Molekularni mehanizmi, ki so odgovorni za insulinsko rezistenco v skeletnem mišičnem vlaknu: insulinska rezistenca in z njo povezana sladkorna bolezen sta zaradi svoje pogostosti v ospredju današnjih medicinskih raziskav. Človeška mišica oživčena in vitro predstavlja odličen poskusni model za raziskave molekularnih mehanizmov, ki privedejo do teh bolezenskih stanj. V sodelovanju s skupino na Inštitutu za patološko fiziologijo, ki razpolaga z najsodobnejšo tehnologijo za spremljanje eksocitotičnih procesov, bomo preučevali molekularne mehanizme, prek katerih insulin sproži translokacijo glukoznih transporterjev 4 (GLUT4)

 
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