Plakins in development and disease

Plakins are large multi-domain molecules that have various functions to link cytoskeletal elements together and to connect them to junctional complexes. Plakins were first identified in epithelial cells where they were found to connect the intermediate filaments to desmosomes and hemidesmosomes [Ruhrberg, C., and Watt, F.M. (1997). The plakin family: versatile organizers of cytoskeletal architecture. Curr Opin Genet Dev 7, 392-397.]. They were subsequently found to be important for the integrity of muscle cells. Most recently, they have been found in the nervous system, where their functions appear to be more complex, including cross-linking of microtubules (MTs) and actin filaments [Leung, C.L., Zheng, M., Prater, S.M., and Liem, R.K. (2001). The BPAG1 locus: Alternative splicing produces multiple isoforms with distinct cytoskeletal linker domains, including predominant isoforms in neurons and muscles. J Cell Biol 154, 691-697., Leung, C.L., Sun, D., Zheng, M., Knowles, D.R., and Liem, R.K. (1999). Microtubule actin cross-linking factor (MACF): a hybrid of dystonin and dystrophin that can interact with the actin and microtubule cytoskeletons. J Cell Biol 147, 1275-1286.]. These plakins have also indicated their relationship to the spectrin superfamily of proteins and the plakins appear to be evolutionarily related to the spectrins, but have diverged to perform different specialized functions. In invertebrates, a single plakin is present in both Drosophila melanogaster and Caenorhabditis elegans, which resemble the more complex plakins found in mammals [Roper, K., Gregory, S.L., and Brown, N.H. (2002). The 'spectraplakins': cytoskeletal giants with characteristics of both spectrin and plakin families. J Cell Sci 115, 4215-4225.]. In contrast, there are seven plakins found in mammals and most of them have alternatively spliced forms leading to a very complex group of proteins with potential tissue specific functions [Jefferson, J.J., Leung, C.L., and Liem, R.K. (2004). Plakins: goliaths that link cell junctions and the cytoskeleton. Nat Rev Mol Cell Biol 5, 542-553.]. In this review, we will first describe the plakins, desmoplakin, plectin, envoplakin and periplakin and then describe two other mammalian plakins, Bullous pemphigoid antigen 1 (BPAG1) and microtubule actin cross-linking factor 1 (MACF1), that are expressed in multiple isoforms in different tissues. We will also describe the relationship of these two proteins to the invertebrate plakins, shortstop (shot) in Drosophila and VAB-10 in C. elegans. Finally, we will describe an unusual mammalian plakin, called epiplakin.     

Mouse models of the laminopathies

The A and B type lamins are nuclear intermediate filament proteins that comprise the bulk of the nuclear lamina, a thin proteinaceous structure underlying the inner nuclear membrane. The A type lamins are encoded by the lamin A gene (LMNA). Mutations in this gene have been linked to at least nine diseases, including the progeroid diseases Hutchinson-Gilford progeria and atypical Werner's syndromes, striated muscle diseases including muscular dystrophies and dilated cardiomyopathies, lipodystrophies affecting adipose tissue deposition, diseases affecting skeletal development, and a peripheral neuropathy. To understand how different diseases arise from different mutations in the same gene, mouse lines carrying some of the same mutations found in the human diseases have been established. We, and others have generated mice with different mutations that result in progeria, muscular dystrophy, and dilated cardiomyopathy. To further our understanding of the functions of the lamins, we also created mice lacking lamin B1, as well as mice expressing only one of the A type lamins. These mouse lines are providing insights into the functions of the lamina and how changes to the lamina affect the mechanical integrity of the nucleus as well as signaling pathways that, when disrupted, may contribute to the disease.     

金皮猪肉质相关组织学特征与CAST基因HinfI多态性的关系研究

本实验选择具有优良肉质的本地金华猪与肉质较差的引进品种皮特兰猪杂交所产的金皮F2代为研究对象,利用PCR-RFLP的方法检测金皮F2代猪CAST基因型的频率,并以肌球蛋白重链（MHC）免疫组织化学、过碘酸希夫反应和苏木精-伊红等多种组织学方法,研究猪背腰最长肌的肌肉组织学特性,并对其进行图像分析,在此基础上采用SAS分析其相互关系。结果发现．PAS染色后肌纤维可区分为三种类型。PAS阴性（Ⅰ）、PAS反应强阳性（Ⅱ-a）和PAS反应强度中间型（Ⅱ-b）。利用肌球蛋白重链单克隆抗体MHCs和MHCf染色结果表明．金皮F2代杂交猪背腰最长肌MHCs阳性（慢肌）纤维的比例为7．62％,快肌纤维为92.38％。CAST基因的扩增产物具有524bp和632bp两个Hinfl多态性片段,定义等位基因为A（114＋192＋400＋632bp）,B（114＋192＋400＋524bp）。AA、BB和AB基因型频率分别为0．1795、0．5897和0．2308,A和B的基因型频率分别为0．4743和0．5256。AA、BB和AB单型对眼肌面积．系水率、pH值、导电率等参数均无显著的影响。CAST基因的HinfI多态性对肌纤维的轴比有显著影响,AA单型有产生较多轴比较大（近似椭圆形）肌纤维的倾向．而BB则控制形成更多的轴比更小（近似于圆形）的肌纤维。具有AA单型的个体具有更多的肌间结缔组织．而具有BB单型的个体的肌间结缔组织较少．AB单型的个体介于两者之间。     

Differential expression of sarcoplasmic proteins in four heterogeneous ovine skeletal muscles

Fiber-type distribution is known to vary widely within and between muscles according to differences in muscle functions. 2-DE and MALDI-MS were used to investigate the molecular basis of muscle fiber type-related variability. We compared four lamb skeletal muscles with heterogeneous fiber-type composition that are relatively rich in fast-twitch fiber types, i.e., the semimembranosus, vastus medialis, longissimus dorsi, and tensor fasciae latae (TL). Our results clearly showed that none of the glycolytic metabolism enzymes detected, including TL which was most strongly glycolytic, made intermuscular differentiation possible. Muscle differentiation was based on the differential expression of proteins involved in oxidative metabolism, including not only citric acid cycle enzymes but also other classes of proteins with functions related to oxidative metabolism, oxidative stress, and probably to higher protein turnover. Detected proteins were involved in transport (carbonate dehydratase, myoglobin, fatty acid-binding protein), repair of misfolding damage (heat shock protein (HSP) 60 kDa, HSP-27 kDa, alpha-crystallin beta subunit, DJ1, stress-induced phosphoprotein), detoxification or degradation of impaired proteins (GST-Pi, aldehyde dehydrogenase, peroxiredoxin, ubiquitin), and protein synthesis (tRNA-synthetase). The fractionating method led to the detection of proteins involved in different functions related to oxidative metabolism that have not previously been shown concomitancy.     

Age-related fatigability in knee extensors and knee flexors during dynamic fatiguing contractions

This study investigated the effects of dynamic knee extension and flexion fatiguing task on torque and neuromuscular responses in young and older individuals. Eighteen young (8 males; 25.1 ± 3.2 years) and 17 older (8 males; 69.7 ± 3.7 years) volunteered. Following a maximal voluntary isometric contraction test, participants performed a fatiguing task involving 22 maximal isokinetic (concentric) knee extension and flexion contractions at 60°/s, while surface EMG was recorded simultaneously from the knee extensors (KE) and flexors (KF). Fatigue-induced relative torque reductions were similar between age groups for KE (peak torque decrease: 25.15% vs 26.81%); however, KF torque was less affected in older individuals (young vs older peak torque decrease: 27.6% vs 11.5%; p < 0.001) and this was associated with greater increase in hamstring EMG amplitude (p < 0.001) and hamstrings/quadriceps peak torque ratio (p < 0.01). Furthermore, KE was more fatigable than KF only among older individuals (peak torque decrease: 26.8% vs 11.5%; p < 0.001). These findings showed that the age-related fatigue induced by a dynamic task was greater for the KE, with greater age-related decline in KE compared to KF.     

Distribution of innervation zone and muscle fiber conduction velocity in the biceps brachii muscle

The spatial distributions of muscle innervation zone (IZ) and muscle fiber conduction velocity (CV) were examined in nine healthy young male participants. High-density surface electromyography (EMG) was collected from the biceps brachii muscle when subjects performed isometric elbow flexions at 20% to 80% of the maximal voluntary contraction (MVC). A total of 9498 samples of IZs were identified and CVs were calculated using the Radon transform. The center and width of IZ sample distribution were compared within four different force levels and six medial to lateral electrode column positions using repeated measures ANOVA and multiple comparison tests. Significant shifts of IZ center were observed in the medial columns (Columns 5, 6, and 7) compared with the lateral columns (Columns 3 and 4) (p < 0.05). Similarly, significant differences in the IZ width were found in Column 7 and 8 compared to Column 3 (p < 0.05). In contrast, muscle CV was unaffected by column position. Instead, muscle CV was faster at 40% and 80% MVC compared to 20% MVC (p < 0.05). The findings of this study add further insights into the physiological properties of the biceps brachii muscle.     

Effects of a tongue training program in Parkinson's disease: Analysis of electrical activity and strength of suprahyoid muscles

ObjectiveTo assess the electrical activity of the suprahyoid muscle and the tongue pressure in a two-wing program of tongue strengthening in individuals with Parkinson's disease (PD).MethodsA pre-post-detraining design study included sixty PD patients assigned to two groups randomly. The experimental group (EG) performed tongue isometric pressure exercises using the Iowa Oral Performance Instrument with an increasing scheme of 5% load weekly and traditional tongue therapy for 8 weeks. The control group (CG) performed only traditional therapy. The electrical activity of suprahyoids was measured using surface electromyography (sEMG) during tongue-to-palate pressure. Four selected domains of the Swallowing Quality of Life Questionnaire (SWAL-QOL) mostly related to tongue strength were considered.ResultsThe experimental group showed increased sEMG values of suprahyoid muscles reaching statistically significant difference at the fourth week of tongue training, while the control did it at the eighth week. Experimental group showed significant improvements in tongue strength (d = 2.128; p = 0.000). Only controls showed detraining effect. Statistically significant difference within groups were found in one and three dimensions of the SWAL-QOL in the CG and EG, respectively.ConclusionAnalysis of electrical activity on suprahyoids muscles provided a better understanding of the changes underlying the outcomes of tongue strength gains obtained through a combined tongue strengthening exercises protocol in PD. Such protocol led not only to increased tongue strength but also to a better perceived swallowing function in PD subjects.     

Low back muscle action potential conduction velocity estimated using high-density electromyography

While a decreasing spectral content of surface electromyography reflects low back muscle fatigue development, reliability of these decreases may be insufficient. Decreasing frequency content is largely determined by decreasing average motor unit action potential conduction velocities (CV), which is considered a more direct measure of muscle fatigue development. However, for the low back muscles it has been proven difficult to identify propagating potentials and consequently estimate the CV. The aim of this study was to estimate the low back muscle CV from high-density multi-channel electromyography by using peak-delay and cross-correlation methods. Fourteen healthy male participants without a history of low-back pain performed a 30 degrees lumbar flexion trial until exhaustion while standing. For 10 out of the 14 participants (118 out of 560 sites) realistic CV estimates were obtained using both methods, the majority likely over the iliocostalis lumborum muscle. Between-method CV differences appeared to be small. Close to the spine a considerable number of sites (79) yielded systematically overestimated low back muscle CV values. Estimating low back muscle CV may allow additional insight into low back muscle fatigue development and potentially improve its monitoring using (high-density) surface electromyography.     

A preliminary muscle activity analysis: Handle based and push-rim wheelchair propulsion

Approximately ninety percent of the wheelchair users worldwide prefer the conventional push rim mode of propulsion for daily mobility and rehabilitation. Even though push-rim wheelchairs help to promote a healthy life style, the high muscular demand and the non-continuous push motions can lead to serious upper extremity injuries. In this study, muscle EMG data of ten healthy subjects were recorded for a newly introduced handle based propulsion mechanism (HBP) and compared to conventional push-rim propulsion at two workloads, 25 W and 35 W respectively. The results for the mean peak muscle activations at both workloads demonstrate that push-rim propulsion leads to higher peak muscle activity compared to HBP at a similar wheelchair forward velocity of 1.11 m/s. The generation of these high peak muscle activations with increasing loads in push-rim propulsion over time can lead to overuse injuries. Overall, the use of the HBP mechanism is less straining to the muscles and may reduce fatigue during prolonged propulsion.