Muscular System Development Research Articles

ADirected influence on the development of the muscular system during the course of kyokushin karate occupations among the teenagers aged 12-15 years with the scoliosis of the first degree

Аннотация Проведено исследование о возможности исправления сколиоза первой степени у подрост-ков в возрасте 12-15 лет в процессе занятий каратэ кёкусинкай при направленном развитии мы-шечного корсета. Используемые средства тренировки включали специально подобранные двига-тельные задания, применяемые в каратэ кёкусинкай, имеющие различную координационную структуру и технику выполнения основных соревновательных действий, связанных с ударными движениями рук и ног. Для эксперимента были сформированы две группы детей в возрасте 12-15 лет со сколиозом первой степени в каждой. Экспериментальная группа детей занималась по мето-дике упражнений разной сложности, применяемых в каратэ кёкусинкай, контрольная по методике, применяемой в адаптивной физической культуре. В результате формирующего педагогического эксперимента было доказано, что развитие силы мышечного корсета туловища выявило положи-тельную тенденцию в исправлении сколиоза подростков. Однако у детей экспериментальной груп-пы, занимающихся по предложенной методике изменения оказались более выраженными, чем у группы детей занимающихся в контрольной группе. Ключевые слова

Is gastrectomy-induced high turnover of bone with hyperosteoidosis and increase of mineralization a typical osteomalacia?

Gastrectomy (GX) is thought to result in osteomalacia due to deficiencies in Vitamin D and Ca. Using a GX rat model, we showed that GX induced high turnover of bone with hyperosteoidosis, prominent increase of mineralization and increased mRNA expression of both osteoclast-derived tartrate-resistant acid phosphatase 5b and osteocalcin. The increased 1, 25(OH)2D3 level and unchanged PTH and calcitonin levels suggested that conventional bone and Ca metabolic pathways were not involved or changed in compensation. Thus, GX-induced bone pathology was different from a typical osteomalacia. Gene expression profiles through microarray analysis and data mining using Ingenuity Pathway Analysis indicated that 612 genes were up-regulated and 1,097 genes were down-regulated in the GX bone. These genes were related functionally to connective tissue development, skeletal and muscular system development and function, Ca signaling and the role of osteoblasts, osteoclasts and chondrocytes. Network analysis indicated 9 genes (Aldehyde dehydrogenase 1 family, member A1; Aquaporin 9; Interleukin 1 receptor accessory protein; Very low density lipoprotein receptor; Periostin, osteoblast specific factor; Aggrecan; Gremlin 1; Angiopoietin-like 4; Wingless-type MMTV integration site family, member 10B) were hubs connected with tissue development and immunological diseases. These results suggest that chronic systemic inflammation might underlie the GX-induced pathological changes in bone.

Deregulation of Specific Sets of Genes Detected by Microarray Analysis of Marrow Stromal Fibroblast Cells Stimulated by IR and VR Light

Conflicting reports have questioned whether low level laser treatment can have an effect on bone. Purpose:The purpose of this study was to examine how light affects gene expression. Methods: Clones of marrow fibroblast stem cells were isolated from human bone marrow and used for primary cell cultures. The cells were grown without light as a control and with light under eight different conditions: two wavelengths of light(830-IR and 633nmVR) at four different energy levels(0.5.1.0,1.5 and 2.0 joules /cm2). Results: Light stimulated some cultures to proliferate up to 40% faster depending on the type of light stimulation using BrdU markers. For the light-stimulated cultures, western blot data for osteocalcin, alkaline phosphatase and RUNX 2, markers of osteoblast differentiation and bone formation, did not show a pattern of increased or early gene expression compared to control cultures. Affymetrix Human exon microarrays of 22,000 protein-encoding genes (40 markers per gene) showed distinct sets of genes being deregulated for each of the eight experimental conditions. A two-fold screen for gene expression differences between control and light-stimulated cultures revealed a complex network of gene expression interactions involving dose response and silencing of inhibitor genes for candidate genes such as RANKL, IL-1A and MMP10. Pathway analysis showed the top VR biological gene networks were skeletal and muscular system development and function, tissue development and amino acid metabolism, consistent with bone turnover. The top biological functions associated with IR stimulation are skin condition, genetic disorders, cancer, immune response. Conclusion: This study provides strong evidence for light stimulating different sets of genes according to wavelength and energy level that can alter bone turnover. The data provides a molecular explanation for the conflicting reports in the current literature.

Evolutional principles of homology in regulatory genes of myogenesis

Analysis of early steps in muscular system development of invertebrates and vertebrates shows that early steps of myogenesis are regulated by genes-orthologs mainly belonging to two families, Pax and bHLH. In the majority of the following organisms, muscles formation (steps of determination and the earliest steps of myogenesis) is regulated by genes orthologs Pax3 which belong to the family Pax: nematodes (Caenorhabditis elegans, Pristionchus pacificus), insects (Drosophila melanogaster), echinoderms (Strongylocentrotus purpuratus), sea squirts (Ciona intestinalis, Holocynthia roretzi), fishes (Danio rerio), amphibians (Xenopus laevis), birds, and mammals (mouse, rat). The nematode C. elegans is an exception since formation of its muscles in this period is regulated by homeobox gene Pal-1 belonging to the family Caudal. The sea squirt C. intestinalis is also an exception because the earliest steps of development involved in further muscle formation are accompanied by activation of the gene CiSna (snail) (gene family basic Zinc finger). The next steps of myogenesis in all analyzed species are regulated by genes orthologs belonging to the family of transcriptional factors bHLH. They along with genes Pax3 are characterized by a high extent of homology in all studied groups of animals.

Identification of gene pathways altered by deletion of the androgen receptor specifically in mineralizing osteoblasts and osteocytes in mice

Androgens play a key role in skeletal growth and maintenance in males and can mediate their actions, at least in part, via the androgen receptor (AR) in osteoblasts. To investigate the mechanisms by which androgens exert their effects via the AR in mineralizing osteoblasts and osteocytes, we identified gene targets/pathways regulated by the AR using targeted gene expression and microarray approaches on bone isolated from mice in which the AR is specifically deleted in mineralizing osteoblasts and osteocytes (mOBL-ARKOs). Gene ontology mining indicated a number of biological processes to be affected in the bones of mOBL-ARKOs including skeletal and muscular system development and carbohydrate metabolism. All genes identified to have altered expression in the bones of mOBL-ARKOs were confirmed by Q-PCR for their androgen responsiveness in an androgen deprivation and replacement mouse model. The osteoblast genes Col1a1 and Bglap and the osteoclast genes Ctsk and RANKL (Tnfs11) were upregulated in the bones of mOBL-ARKOs, consistent with the increased matrix synthesis, mineralization, and bone resorption observed previously in these mice. Of significant interest, we identified genes involved in carbohydrate metabolism (adiponectin and Dpp4) and in growth and development (GH, Tgfb (Tgfb2), Wnt4) as potential targets of androgen action via the AR in mineralizing osteoblasts.

Genome-wide association of an integrated osteoporosis-related phenotype: is there evidence for pleiotropic genes?

Multiple musculoskeletal traits assessed by various methods at different skeletal sites serve as surrogates for osteoporosis risk. However, it is a challenge to select the most relevant phenotypes for genetic study of fractures. Principal component analyses (PCA) were conducted in participants of the Framingham Osteoporosis Study on 17 measures including bond mineral density (BMD) (hip and spine), heel ultrasound, leg lean mass (LLM), and hip geometric indices, adjusting for covariates (age, height, body mass index [BMI]), in a combined sample of 1180 men and 1758 women, as well as in each sex. Four principal components (PCs) jointly explained ~69% of the total variability of musculoskeletal traits. PC1, explaining ~33% of the total variance, was referred to as the component of "Bone strength," because it included the hip and spine BMD as well as several hip cross-sectional properties. PC2 (20.5% variance) was labeled as "Femoral cross-sectional geometry;" PC3 (~8% variance) captured only ultrasound measures; PC4, explaining ~7% variance, was correlated with LLM and hip geometry. We then evaluated ~2.5 mil SNPs for association with PCs 1, 2, and 4. There were genome-wide significant associations (p < 5 × 10⁻⁸) between PC2 and HTR1E (that codes for one of the serotonin receptors) and PC4 with COL4A2 in women. In the sexes-combined sample, AKAP6 was associated with PC2 (p = 1.40 × 10⁻⁷). A single nucleotide polymorphism (SNP) in HTR1E was also associated with the risk of nonvertebral fractures in women (p = 0.005). Functions of top associated genes were enriched for the skeletal and muscular system development (p < 0.05). In conclusion, multivariate combination provides genetic associations not identified in the analysis of primary phenotypes. Genome-wide screening for the linear combinations of multiple osteoporosis-related phenotypes suggests that there are variants with potentially pleiotropic effects in established and novel pathways to be followed up to provide further evidence of their functions.

Meta-analysis of gene-based genome-wide association studies of bone mineral density in Chinese and European subjects

SummaryGene-based association approach could be regarded as a complementary analysis to the single SNP association analysis. We meta-analyzed the findings from the gene-based association approach using the genome-wide association studies (GWAS) data from Chinese and European subjects, confirmed several well established bone mineral density (BMD) genes, and suggested several novel BMD genes.IntroductionThe introduction of GWAS has greatly increased the number of genes that are known to be associated with common diseases. Nonetheless, such a single SNP GWAS has a lower power to detect genes with multiple causal variants. We aimed to assess the association of each gene with BMD variation at the spine and hip using gene-based GWAS approach.MethodsWe studied 778 Hong Kong Southern Chinese (HKSC) women and 5,858 Northern Europeans (dCG); age, sex, and weight were adjusted in the model. The main outcome measure was BMD at the spine and hip.ResultsNine genes showed suggestive p value in HKSC, while 4 and 17 genes showed significant and suggestive p values respectively in dCG. Meta-analysis using weighted Z-transformed test confirmed several known BMD genes and suggested some novel ones at 1q21.3, 9q22, 9q33.2, 20p13, and 20q12. Top BMD genes were significantly associated with connective tissue, skeletal, and muscular system development and function (p < 0.05). Gene network inference revealed that a large number of these genes were significantly connected with each other to form a functional gene network, and several signaling pathways were strongly connected with these gene networks.ConclusionOur gene-based GWAS confirmed several BMD genes and suggested several novel BMD genes. Genetic contribution to BMD variation may operate through multiple genes identified in this study in functional gene networks. This finding may be useful in identifying and prioritizing candidate genes/loci for further study.Electronic supplementary materialThe online version of this article (doi:10.1007/s00198-011-1779-7) contains supplementary material, which is available to authorized users.

Specific and extensive endometrial deregulation is present before conception in IVF/ICSI repeated implantation failures (IF) or recurrent miscarriages

The objective was to examine if IVF/ICSI repeated implantation failures (IF) or recurrent miscarriages (RM) could be related to preconceptional endometrial deregulations. IF was defined as the absence of pregnancy despite the transfer of at least ten IVF/ICSI good quality embryos, and RM as having at least three unexplained miscarriages. Fertile controls (FC) were women who had given birth at least once. Endometrial biopsy was performed in the mild luteal phase of a non-conceptual cycle (five women were selected in each group). Affymetrix chips (GeneChip Human Genome U133 Plus2.0 Array) were used for hybridization. Data were normalized by the gcRMA method, and raw p values adjusted by the Bonferroni procedure (1%). Differential expression of selected genes was analysed using real-time PCR. Gene networks and biological functions were explored using the Ingenuity Pathways Analysis software. Endometrial gene expression profiles at the time of uterine receptivity differ dramatically in the endometrium among FC, RM, and IF patients. Compared to FC, 2126 and 2477 genes are differentially expressed in IF and RM groups, respectively, and 2363 between IF and RM. In both conditions, differential gene expression referred mainly to DNA transcription and expression. Other main cellular functions deregulated in IF conditions correspond to cell morphology, cellular development, cell cycle, and cellular assembly, while in RM conditions, deregulated cellular functions relate to cell signalling (degradation of cyclic AMP and calcium metabolism) and cellular maintenance. In both conditions, there is an over-representation of deregulations related to the haematological system. In the IF condition, cell-mediated immune response and nervous system development and function are highly deregulated, while in RM patients, main deregulations are in organ and tissue development, humoral immune response, and muscular system development and function. Extensive endometrial deregulations are present before conception in patients who experienced IF or RM with both distinct and common deregulation.

Abstract 4930: An integrated proteomic and bioinformatic approach identifies a subset of EGFR-expressing epithelial ovarian cancer

Abstract Epithelial Ovarian Cancer (EOC) is initially characterized by intraperitoneal growth of tumor masses followed by an accumulation of ascitic fluid containing tumor cells, inflammatory leukocytes, cytokines and chemokines. Among those latter, IL-6 and IL-8 have been found to contribute to cell proliferation of EOC. EGFR, detected in up to 60% of EOCs, is associated with poor outcome. EGFR overexpression has been also associated to chemotherapy resistance. However, treatment of EOC patients with anti-EGFR resulted to give very poor response. In this study we show that EGF stimulation of EOC lines IGROV1 and OAW42, displaying comparable high expression levels of EGFR, induced, in conditioned medium of both cell lines production of several cyto/chemokines as detected by Bioplex analysis. IL-6, PAI-1 and IL-8 were however specifically produced upon EGFR activation only in IGROV1 and, conversely, silencing of EGFR in this cell line abrogated, following stimulation, their induction. Immunohistochemistry of EOC tissue samples with an anti-P(Y1173)-EGFR and dosage of IL-6 in the corresponding ascitic fluids indicated that, although not significant, IL-6 levels trended upwards with P-EGFR expression. Moreover in the ascitic fluids from EOC patients IL-6 and PAI-1 significantly correlated (Spearman correlation: r=0.79). In order to elucidate the molecular determinants of EGFR-, IL-6-, PAI-1- and IL-8-expressing EOC, a computational analysis was performed on the gene expression profile of 204 serous EOC patients contained in a public data set downloaded from GEO repository (GSE9891). Pearson correlation analyses were performed individually on EGFR, IL6, IL8 and PAI-1 (SERPINE1) expression intensities versus intensity of all the other genes in the data set. Correlation values and p-values of each pair were computed and p-values were adjusted for simple multiple testing procedures (“Bonferroni” and others) through the R programming language. This analysis yielded lists of candidate genes positively or negatively correlated to those of interest and, among those, 39 were in common among the genes correlated to IL6 and PAI-1. These genes were loaded in Ingenuity Pathway software and 23 genes resulted to be directly or indirectly associated with the “Cellular Movement, Embryonic Development, Cellular Growth and Proliferation” and “Skeletal and Muscular System Development and Function, Tissue Development” with the highest scores. Overall we identified a subtype of EGFR-expressing EOCs showing a particular signature of microenvironment-related genes thus facilitating the development of alternative approaches potentially suited for improving diagnosis and treatment of EOCs. Partially supported by Italian Health Ministry (Grant ACC1) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4930. doi:10.1158/1538-7445.AM2011-4930

Genome-wide expression profiling of urinary bladder implicates desmosomal and cytoskeletal dysregulation in the bladder exstrophy-epispadias complex

The bladder exstrophy-epispadias complex (BEEC) represents a spectrum of urological abnormalities where part, or all, of the distal urinary tract fails to close during development, becoming exposed on the outer abdominal wall. While the etiology of BEEC remains unknown, strong evidence exists that genetic factors are implicated. To understand the pathways regulating embryonic bladder development and to identify high-probability BEEC candidate genes, we conducted a genome-wide expression profiling (GWEP) study using normal and exstrophic human urinary bladders and human and mouse embryologic bladder-precursor tissues. We identified 162 genes differentially expressed in both embryonic and postnatal human samples. Pathway analysis of these genes revealed 11 biological networks with top functions related to skeletal and muscular system development, cellular assembly and development, organ morphology, or connective tissue disorders. The two most down-regulated genes desmin (DES, fold-change, -74.7) and desmuslin (DMN, fold-change, -53.0) are involved in desmosome mediated cell-cell adhesion and cytoskeletal architecture. Intriguingly, the sixth most overexpressed gene was desmoplakin (DSP, fold-change, +48.8), the most abundant desmosomal protein. We found 30% of the candidate genes to be directly associated with desmosome structure/function or cytoskeletal assembly, pointing to desmosomal and/or cytoskeletal deregulation as an etiologic factor for BEEC. Further findings indicate that p63, PERP, SYNPO2 and the Wnt pathway may also contribute to BEEC etiology. This study provides the first expression profile of urogenital genes during bladder development and points to the high-probability candidate genes for BEEC.

A comparative transcriptomic study of vitamin E and an algae-based antioxidant as antioxidative agents: Investigation of replacing vitamin E with the algae-based antioxidant in broiler diets

Previous study indicated that inclusion of an algae-based antioxidant as an antioxidative agent [EconomasE, Alltech, Nicholasville, KY; EcoE] significantly reduced the amount of vitamin E (VE) required in broiler diets without compromising performance and meat quality. To assess the mechanisms related to the VE-saving activity of EcoE, as well as other potential functions related to EcoE and VE supplementation, we analyzed gene expression profiles of breast muscle from broilers fed a control diet, the control diet + 50 IU of VE/kg, the control diet + 100 IU of VE/kg, or the control diet + 200 g of EcoE/ton. Evaluation of the serum antioxidant capacity indicated that dietary supplementation of either a high level of VE (50 or 100 IU of VE/kg) or EcoE significantly improved bird antioxidant status. Analysis of gene expression profiles indicated that expression of 542 genes of the breast muscle were altered (P < 0.05, fold change >1.2) by dietary treatments, of which a significant part were commonly regulated by EcoE and VE (especially the control diet + 50 IU of VE/kg). In addition to the process of cellular oxidation, gene ontology analysis indicated the involvement of EcoE and VE on cell morphology, skeletal and muscular system development and function, immune response, and multiple metabolic processes, including lipid, carbohydrate, and drug metabolism. Results of this experiment indicate that the biological roles of high VE, including its activity as an antioxidant, can be greatly mimicked at the transcriptional level by EcoE, and they suggest a relationship of functional redundancy between VE and EcoE in the broiler diets.

Gene expression profile of the fibrotic response in the peritoneal cavity

The cellular response to materials implanted in the peritoneal cavity has been utilised to produce tissue for grafting to hollow smooth muscle organs (blood vessels, bladder, uterus and vas deferens). To gain insight into the regulatory mechanisms involved in encapsulation of a foreign object, and subsequent differentiation of encapsulating cells, the present study used microarray technology and real-time RT-PCR to identify the temporal changes in gene expression associated with tissue development. Immunohistochemical analysis showed that 3–7 days post-implantation of foreign objects (cubes of boiled egg white) into rats, they were encapsulated by tissue comprised primarily of haemopoietic (CD45 +) cells, mainly macrophages (CD68 +, CCR1 +). By day 14, tissue capsule cells no longer expressed CD68, but were positive for myofibroblast markers α-smooth muscle (SM) actin and SM22. In accordance with these results, gene expression data showed that early capsule (days 3–7) development was dominated by the expression of monocyte/macrophage-specific genes ( CD14, CSF-1, CSF-1R, MCP-1) and pro-inflammatory mediators such as transforming growth factor ( TGF- β). As tissue capsule development progressed (days 14–21), myofibroblast-associated and pro-fibrotic genes (associated with TGF-β and Wnt/β-catenin signalling pathways, including Wnt 4, TGFβRII, connective tissue growth factor (CTGF), SMADs-1, -2, -4 and collagen-1 subunits) were significantly up-regulated. The up-regulation of genes associated with Cardiovascular and Skeletal and Muscular System Development at later time-points suggests the capacity of cells within the tissue capsule for further differentiation to smooth muscle, and possibly other cell types. The identification of key regulatory pathways and molecules associated with the fibrotic response to implanted materials has important applications not only for optimising tissue engineering strategies, but also to control deleterious fibrotic responses.

Genome expression profiling and network analysis of nitrite therapy during chronic ischemia: Possible mechanisms and interesting molecules

Sodium nitrite is widely recognized to be a highly effective NO donor for the treatment of several ischemic tissue disorders. However, mechanisms by which nitrite confers cytoprotection during ischemic disorders remain largely unknown. In this study, we used genome expression profiling approaches to evaluate changes in gene expression in the hind-limb ischemia model using vehicle or sodium nitrite therapy. Sodium nitrite significantly restored ischemic tissue perfusion by day 3 post-ligation which returned to normal by day 7. Genesifter analysis of Affymetrix GeneChip data revealed a significant down-regulation of gene expression profiles at day 3, whereas gene expression profiles were predominantly up-regulated at day 7. Ingenuity network analysis of gene expression profiles at day 3 showed a strong decrease in gene expression from networks associated with immune functions such as acute inflammatory responses, antigen presentation, and humoral immune responses while networks containing increased gene expression profiles were associated with cardiovascular, skeletal, and muscle system development and function. Network analysis of day 7 gene array data revealed predominant up-regulation of genes associated with cell survival, tissue morphology, connective tissue function, skeletal and muscular system development, and lymphoid tissue structure and development. These data suggest that sodium nitrite elicits potent anti-inflammatory and pro-angiogenic gene responses at early time points which is later followed by up-regulation of genes associated with tissue repair and homeostasis.

Carbon Monoxide Modulates α–Smooth Muscle Actin and Small Proline Rich-1a Expression in Fibrosis

Carbon monoxide (CO) is a biologically active molecule produced in the body by the stress-inducible enzyme, heme oxygenase. We have previously shown that CO suppresses fibrosis in a murine bleomycin model. To investigate the mechanisms by which CO opposes fibrogenesis, we performed gene expression profiling of fibroblasts treated with transforming growth factor-beta(1) and CO. The most highly differentially expressed categories of genes included those related to muscular system development and the small proline-rich family of proteins. We confirmed in vitro, and in an in vivo bleomycin model of lung fibrosis, that CO suppresses alpha-smooth muscle actin expression and enhances small proline-rich protein-1a expression. We further show that these effects of CO depend upon signaling via the extracellular signal-regulated kinase pathway. Our results demonstrate novel transcriptional targets for CO and further elucidate the mechanism by which CO suppresses fibrosis.

Rotifer muscles as revealed by phalloidin‐TRITC staining and confocal scanning laser microscopy

AbstractBy combining phalloidin‐TRITC staining with confocal scanning laser microscopy (CSLM), the pattern of the musculature in two species of Rotifera, Euchlanis dilatata unisetata and Brachionus quadridentatus is revealed. The same general muscle pattern prevails in both species. The major components of the body wall musculature are: 1. retractor muscles (5 pairs in E. dilatata unisetata and 3 pairs in B. quadridentatus); 2. Two pairs of dorso‐ventral muscles; 3. Two pairs of perpendicular muscles (in E. dilatata unisetata); 4. retractors of the corona (median, lateral and ventral); 5. Foot retractors. In addition, three pairs of cutaneo‐visceral muscles and visceral muscles (including mastax muscles) are described. The sphincter of the corona was found only in B. quadridentatus. The high degree of muscle differentiation points to a high level of development of rotifer muscular system.