Izražanje genskih prepisov za izooblike težkih verig miozina v različnih tipih mišičnih vlaken človeka = Expression of myosin heavy chain transcripts with distinct types of human muscle fibres : doktorska disertacija
Smerdu, Vika
While four genes determine four major isoforms (I, IIA, IIX and IIB) of myosinheavy chains (MyHC) in adult rodent skeletal muscles, only three isoforms, MyHC-I, -IIA and -IIB, were found in mature ...human skeletal muscles. At the beginning of this study three human MyHC genes, coding for MyHC-I, embryonic and perinatal MyHC isoforms, had already been determined. It was known that two additional MyHC genes also exist, they were supposed to be expressed in type II fibres. Two clones from human MyHC cDNA library were found to be very similar to cDNA clones for MyHC-IIA and -IIX of rat. Thus it was presumed that they correspond to the two undefined genes on chromosome 17 and that they are expressed in type II fibres of adult human skeletal muscles.To verify this assumption, the expression of MyHC transcripts (mRNA), corresponding to the two clones and to the three known MyHC genes, were analysed for the first time in this study. The MyHC transcript expression was revealed by in situ hybridisation technique with specific radiolabelled RNA probes. Beside embryonic, perinatal and beta probe also two probes, corresponding to the two partly known cDNA clones, were synthesised by transcription in vitro technique. Considering high homology of the two cDNA clones with rat cDNA clones for MyHC-IIA and -IIX, the corresponding probes were designated as 2a and 2x probes. According to the pattern of MyHC transcript expression fibre groups were determined and correlated to histochemical phenotypes of fibres (type I, IIC, IIA, IIAB and IIB), demonstrated with histochemical reaction for myofibrillar adenosine triphosphatase (ATPase). To show MyHC isoform expression monoclonal antibodiesspecific for rat MyHC were used and metabolic profiles of fibres were determined according to the activity of oxidative enzyme nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) and glycolytic enzyme a-glycerophosphate dehydrogenase (a-GPDH). (Abstract truncated at 2000 characters).
Type of material - dissertation
; adult, serious
Publication and manufacture - Ljubljana : [V. Smerdu], 2002
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