Analysis of Embryonic and Larval Zebrafish Skeletal Myofibers from Dissociated Preparations

The zebrafish has  proven to be a valuable model system for exploring skeletal muscle function and  for studying human muscle diseases. Despite  the many ...

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

The zebrafish model is an emerging system for the study of neuromuscular disorders.  In the study of neuromuscular diseases, the integrity of the muscle ...

University of Michigan 

The Hospital for Sick Children

The University of Toronto

Transposons have been used in invertebrates for transgenesis and insertional mutagens in genetic screens. We tested a functional transposon called Sleeping Beauty in the one-cell mouse embryo. In this report, we describe experiments in which transposon vectors were injected into one-cell mouse embryos with mRNA expressing the SB10 transposase enzyme. Molecular evidence of transposition was obtained by cloning of insertion sites from multiple transgenic mice produced by SB10 mRNA/transposon coinjection. We also demonstrate germ-line transmission and expression from transposed elements. This technique has promise as a germ-line transgenesis method in other vertebrate species and for insertional mutagenesis in the mouse.

Mammalian germ-line transgenesis by transposition.

We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish (Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.

Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon.

Among functional elements of a metazoan gene, enhancers are particularly difficult to find and annotate. Pioneering experiments in Drosophila have demonstrated the value of enhancer "trapping" using an invertebrate to address this functional genomics problem.

Enhancer trapping in zebrafish using the Sleeping Beauty transposon.

Sleeping Beauty transposon for efficient gene delivery.

We report a novel gene tagging, identification and mutagenicity ('gene-breaking') method for the zebrafish, Danio rerio. This modular approach consists of two distinct and separable molecular cassettes. The first is a gene-finding cassette. In this study, we employed a 3' gene-tagging approach that selectively 'traps' transcripts regardless of expression status, and we show that this cassette identifies both known and novel endogenous transcripts in transgenic zebrafish. The second is a transcriptional termination mutagenicity cassette assembled from a combination of a splice acceptor and polyadenylation signal to disrupt tagged transcripts upon integration into intronic sequence. We identified both novel and conserved loci as linked phenotypic mutations using this gene-breaking strategy, generating molecularly null mutations in both larval lethal and adult viable loci. We show that the Histone 2a family member z (H2afza) variant is essential for larval development through the generation of a lethal locus with a truncation of conserved carboxy-terminal residues in the protein. In principle this gene-breaking strategy is scalable for functional genomics screens and can be used in Sleeping Beauty transposon and other gene delivery systems in the zebrafish.

Gene-breaking transposon mutagenesis reveals an essential role for histone H2afza in zebrafish larval development.

Use of the Sleeping Beauty transposon system for stable gene expression in mouse embryonic stem cells.

Congenital myopathies are clinically and genetically heterogeneous diseases that typically present in childhood with hypotonia and weakness and are most commonly defined by changes observed in muscle biopsy. Approximately 40% of congenital myopathies are currently genetically unresolved. We identified a family with dominantly inherited congenital myopathy characterized by distal weakness and biopsy changes that included core-like areas and increased internalized nuclei. To identify the causative genetic abnormality in this family, we performed linkage analysis followed by whole-exome capture and next-generation sequencing. A splice-acceptor variant in previously uncharacterized CCDC78 was detected in affected individuals and absent in unaffected family members and > 10,000 controls. This variant alters RNA-transcript processing and results in a 222Â bp in-frame insertion. CCDC78 is expressed in skeletal muscle, enriched in the perinuclear region and the triad, and found in intracellular aggregates in patient muscle. Modeling of the CCDC78 mutation in zebrafish resulted in changes mirroring the human disease that included altered motor function and abnormal muscle ultrastructure. Using a combination of linkage analysis, next-generation sequencing, and modeling in the zebrafish, we have identified a CCDC78 mutation associated with a unique myopathy with prominent internal nuclei and atypical cores.

Dominant mutation of CCDC78 in a unique congenital myopathy with prominent internal nuclei and atypical cores.

Myopathies cong&eacute;nitales sont cliniquement et g&eacute;n&eacute;tiquement h&eacute;t&eacute;rog&egrave;nes maladies g&eacute;n&eacute;ralement pr&eacute;sent dans l&#39;enfance avec hypotonie et faiblesse et plus commun&eacute;ment d&eacute;finies par les changements observ&eacute;s dans la biopsie musculaire. Environ 40 % des myopathies cong&eacute;nitales sont actuellement g&eacute;n&eacute;tiquement non r&eacute;solue. Nous avons identifi&eacute; une famille avec dominance h&eacute;r&eacute;ditaire myopathie cong&eacute;nitale caract&eacute;ris&eacute;e par des changements biopsie et faiblesse distales qui inclus les domaines nucl&eacute;aire et a augment&eacute; les noyaux int&eacute;rioris&eacute;es. Pour identifier l&#39;anomalie g&eacute;n&eacute;tique causal dans cette famille, nous avons effectu&eacute; analyse de liaison suivie d&#39;ensemble-l&#39;exome capture et s&eacute;quen&ccedil;age de nouvelle g&eacute;n&eacute;ration. Une variante de l&#39;accepteur d&#39;&eacute;pissage en CCDC78 non d&eacute;finies auparavant a &eacute;t&eacute; d&eacute;tect&eacute;e dans touch&eacute;s particuliers et absents chez les membres de la famille affect&eacute;es et &gt; 10 000 contr&ocirc;les. Cette variante modifie le traitement transcrit et se traduit par une insertion dans le cadre de bp &agrave; l&#39;adresse 222. CCDC78 est exprim&eacute;e dans le muscle squelettique, enrichi dans la r&eacute;gion p&eacute;rinucl&eacute;aire et la triade et trouv&eacute; dans les agr&eacute;gats intracellulaires dans les muscles de patients. Mod&eacute;lisation de la CCDC78 mutation chez le poisson z&egrave;bre a entra&icirc;n&eacute; des modifications de mise en miroir de la maladie humaine qui comprenait modifi&eacute; la fonction motrice et l&#39;ultrastructure de musculaire anormale. En utilisant une combinaison de linkage analysis, g&eacute;n&eacute;ration de s&eacute;quen&ccedil;age et mod&eacute;lisation dans le poisson-z&egrave;bre, nous avons identifi&eacute; une mutation de CCDC78 associ&eacute;e &agrave; une myopathie unique avec des noyaux internes et noyaux atypiques.

Mutation dominante de CCDC78 dans une myopathie cong&eacute;nitale unique avec des noyaux internes et noyaux atypiques.

Le miopatie congenite sono malattie clinicamente e geneticamente eterogenee che tipicamente presentano nell&#39;infanzia con ipotonia e debolezza e sono pi&ugrave; comunemente definite dai cambiamenti osservati nella biopsia muscolare. Attualmente circa il 40% delle miopatie congenite sono geneticamente non risolti. Abbiamo identificato una famiglia con miopatia congenita dominante, caratterizzata da cambiamenti biopsia e debolezza distale che includeva core-come le zone e interiorizzati nuclei &egrave; aumentato. Per identificare l&#39;anomalia genetica causativo in questa famiglia, abbiamo eseguito le analisi di linkage seguita da acquisizione intero-exome e sequenziamento di nuova generazione. Una variante di splice-accettore in CCDC78 precedentemente atipici &egrave; stata rilevata interessati e assente nei controlli &amp;gt; 10.000 e membri della famiglia inalterati. Questa variante altera elaborazione RNA-trascrizione e si traduce in un 222 inserimento nel telaio bp. CCDC78 &egrave; espressa nel muscolo scheletrico, arricchito nella regione perinucleare e la triade e trovato in aggregati intracellulari nel paziente muscolare. Modellazione di CCDC78 mutazione nello zebrafish ha provocato cambiamenti la malattia umana che comprendeva il mirroring alterata funzione motoria e ultrastruttura muscolare anormale. Utilizzando una combinazione di analisi di linkage, next-generation sequencing e modeling in the zebrafish, abbiamo identificato una mutazione di CCDC78 associata con un&#39;unica miopatia con nuclei prominenti interni e nuclei atipici.

Mutazione dominante di CCDC78 in un&#39;unica miopatia congenita con nuclei prominenti interni e nuclei atipici.

The Î²-tropomyosin gene encodes a component of the sarcomeric thin filament. Rod-shaped dimers of tropomyosin regulate actin-myosin interactions and Î²-tropomyosin mutations have been associated with nemaline myopathy, cap myopathy, Escobar syndrome and distal arthrogryposis types 1A and 2B. In this study, we expand the allelic spectrum of Î²-tropomyosin-related myopathies through the identification of a novel Î²-tropomyosin mutation in two clinical contexts not previously associated with Î²-tropomyosin. The first clinical phenotype is core-rod myopathy, with a Î²-tropomyosin mutation uncovered by whole exome sequencing in a family with autosomal dominant distal myopathy and muscle biopsy features of both minicores and nemaline rods. The second phenotype, observed in four unrelated families, is autosomal dominant trismus-pseudocamptodactyly syndrome (distal arthrogryposis type 7; previously associated exclusively with myosin heavy chain 8 mutations). In all four families, the mutation identified was a novel 3-bp in-frame deletion (c.20_22del) that results in deletion of a conserved lysine at the seventh amino acid position (p.K7del). This is the first mutation identified in the extreme N-terminus of Î²-tropomyosin. To understand the potential pathogenic mechanism(s) underlying this mutation, we performed both computational analysis and in vivo modelling. Our theoretical model predicts that the mutation disrupts the N-terminus of the Î±-helices of dimeric Î²-tropomyosin, a change predicted to alter protein-protein binding between Î²-tropomyosin and other molecules and to disturb head-to-tail polymerization of Î²-tropomyosin dimers. To create an in vivo model, we expressed wild-type or p.K7del Î²-tropomyosin in the developing zebrafish. p.K7del Î²-tropomyosin fails to localize properly within the thin filament compartment and its expression alters sarcomere length, suggesting that the mutation interferes with head-to-tail Î²-tropomyosin polymerization and with overall sarcomeric structure. We describe a novel Î²-tropomyosin mutation, two clinical-histopathological phenotypes not previously associated with Î²-tropomyosin and pathogenic data from the first animal model of Î²-tropomyosin-related myopathies.

Novel deletion of lysine 7 expands the clinical, histopathological and genetic spectrum of TPM2-related myopathies.

Le g&egrave;ne de la &beta;-tropomyosine code un composant du filament mince du sarcom&egrave;re. B&acirc;tonnet de dim&egrave;res de tropomyosine r&eacute;gulent les interactions actine-myosine et &beta;-tropomyosine mutations ont &eacute;t&eacute; associ&eacute;es nemaline myopathy, cap myopathie, syndrome d&#39;Escobar et arthrogrypose distale type 1 a et 2 b. Dans cette &eacute;tude, nous &eacute;largissons la gamme all&eacute;lique des myopathies &beta;-tropomyosine-li&eacute;s par le biais de l&#39;identification d&#39;une mutation nouvelle &beta;-tropomyosine dans deux contextes cliniques n&#39;a pas &eacute;t&eacute; associ&eacute; &agrave; la &beta;-tropomyosine. Le premier ph&eacute;notype clinique est myopathie core-tige, avec une mutation de &beta;-tropomyosine d&eacute;couverte par s&eacute;quen&ccedil;age de l&#39;exome ensemble dans une famille avec autosomiques dominantes distales myopathie et muscle biopsie caract&eacute;ristiques des tiges les mini-carottes et nemaline. Le ph&eacute;notype deuxi&egrave;me, observ&eacute; dans quatre familles ind&eacute;pendantes, est autosomique dominante trismus-pseudocamptodactyly syndrome (arthrogrypose distale type 7; pr&eacute;c&eacute;demment associ&eacute;s exclusivement &agrave; des mutations de la cha&icirc;ne lourde 8 la myosine). Dans toutes les familles de quatre, la mutation identifi&eacute;e &eacute;tait une nouvelle suppression de 3-bp dans le cadre (c.20_22del) qui se traduit par la suppression d&#39;une lysine conserv&eacute;e &agrave; la septi&egrave;me position d&#39;acides amin&eacute;s (p.K7del). Il s&#39;agit de la premi&egrave;re mutation identifi&eacute;e dans l&#39;extr&ecirc;me N-terminale de la &beta;-tropomyosine. Pour comprendre les m&eacute;canismes pathog&egrave;nes potentiels qui sous-tendent cette mutation, nous avons effectu&eacute; l&#39;analyse de calcul et de mod&eacute;lisation in vivo. Notre mod&egrave;le th&eacute;orique pr&eacute;dit que la mutation perturbe l&#39;extr&eacute;mit&eacute; N-terminale des h&eacute;lices-&alpha; &beta; dim&egrave;res-tropomyosine, un changement pr&eacute;vu de modifier la liaison de prot&eacute;ines entre &beta;-tropomyosine et d&#39;autres mol&eacute;cules et perturber la polym&eacute;risation de dim&egrave;res de &beta;-tropomyosine t&ecirc;te &agrave; queue. Pour cr&eacute;er un mod&egrave;le in vivo, nous avons exprim&eacute; des &beta;-tropomyosine sauvage ou p.K7del dans le pays en d&eacute;veloppement chez le poisson z&egrave;bre. p.K7del &beta;-tropomyosine ne parvient pas &agrave; localiser correctement dans le compartiment de filaments minces et son expression modifie la longueur du sarcom&egrave;re, sugg&eacute;rant que la mutation interf&egrave;re avec la polym&eacute;risation de t&ecirc;te-&agrave;-queue &beta;-tropomyosine et avec la structure globale du sarcom&egrave;re. Nous d&eacute;crivons une mutation nouvelle &beta;-tropomyosine, deux ph&eacute;notypes cliniques et histopathologiques, n&#39;a pas &eacute;t&eacute; associ&eacute;s &agrave; la &beta;-tropomyosine et donn&eacute;es pathog&egrave;nes depuis le premier mod&egrave;le animal de myopathies li&eacute;es &agrave; &beta;-tropomyosine.

Nouvelle suppression de lysine 7 &eacute;largit le spectre clinique, histopathologique et g&eacute;n&eacute;tique des myopathies li&eacute;es TPM2.

Il &beta;-tropomiosina gene codifica per un componente del sarcomero filamento sottile. Asta-a forma di dimeri di tropomiosina regolano interazione actina-miosina e tropomiosina &beta; mutazioni sono state associate con miopatia nemalinica, cap miopatia, sindrome di Escobar e artrogriposi distale tipo 1A e 2B. In questo studio, abbiamo espandere lo spettro allelica di miopatie &beta;-tropomiosina-correlati attraverso l&#39;identificazione di una mutazione del romanzo &beta;-tropomiosina in due contesti clinici non precedentemente associati con &beta;-tropomiosina. Il primo fenotipo clinico &egrave; miopatia del core-asta, con una mutazione &beta;-tropomiosina scoperta mediante sequenziamento exome intero in una famiglia con trasmissione autosomiche dominante distale miopatia e muscolo biopsia caratteristiche di canne sia minicores e nemalinica. Il fenotipo secondo, osservato in quattro famiglie indipendenti, &egrave; la sindrome autosomica dominante Trisma-pseudocamptodattilia (artrogriposi distale, tipo 7; precedentemente associato esclusivamente a mutazioni 8 catena pesante della miosina). In tutte le quattro famiglie, la mutazione identificata era una romanzo 3-bp-struttura di eliminazione (c.20_22del) che si traduce nell&#39;eliminazione di una lisina conservata presso la settima posizione dell&#39;amminoacido (p.K7del). Questa &egrave; la prima mutazione identificata in estrema N-terminus di &beta;-tropomiosina. Per comprendere i potenziali meccanismi patogeni comprendono sottostante questa mutazione, abbiamo eseguito analisi computazionale e per la modellazione in vivo. Il nostro modello teorico predice che la mutazione sconvolge il N-terminus di &alpha;-eliche di dimerica &beta;-tropomiosina, un cambiamento previsto per alterare il legame della proteina-proteina tra &beta;-tropomiosina e altre molecole e disturbare la testa-coda la polimerizzazione dei dimeri &beta;-tropomiosina. Per creare un modello in vivo, abbiamo espresso la wild-type o p.K7del &beta;-tropomiosina in the zebrafish in via di sviluppo. p.K7del &beta;-tropomiosina non riesce a localizzare correttamente all&#39;interno del vano del filamento sottile e sua espressione altera la lunghezza del sarcomero, suggerendo che la mutazione interferisce con polimerizzazione di testa-coda &beta;-tropomiosina e struttura nel complesso sarcomero. Descriviamo una mutazione del romanzo &beta;-tropomiosina, due fenotipi clinici istopatologici non precedentemente associati con &beta;-tropomiosina e patogeni dati dal primo modello animale di miopatie &beta;-tropomiosina-correlati.

Romanzo cancellazione di lisina 7 amplia lo spettro clinico, istopatologico e genetico di miopatie TPM2-correlati.

Dynamin-2 (DNM2) is a large GTPase involved in clathrin-mediated endocytosis and related trafficking pathways. Mutations in human DNM2 cause two distinct neuromuscular disorders: centronuclear myopathy and Charcot-Marie-Tooth disease. Zebrafish have been shown to be an excellent animal model for many neurologic disorders, and this system has the potential to inform our understanding of DNM2-related disease. Currently, little is known about the endogenous zebrafish orthologs to human DNM2. In this study, we characterize two zebrafish dynamin-2 genes, dnm2 and dnm2-like. Both orthologs are structurally similar to human DNM2 at the gene and protein levels. They are expressed throughout early development and in all adult tissues examined. Knockdown of dnm2 and dnm2-like gene products resulted in extensive morphological abnormalities during development, and expression of human DNM2 RNA rescued these phenotypes. Our findings suggest that dnm2 and dnm2-like are orthologs to human DNM2, and that they are required for normal zebrafish development.

Two dynamin-2 genes are required for normal zebrafish development.

Dynamine-2 (DNM2) est une GTPase grande impliqu&eacute;s dans l&#39;endocytose clathrine et associ&eacute;s &agrave; des voies de trafic. Mutations de DNM2 humaine causent deux troubles neuromusculaires distinctes : centronuclear myopathy et maladie de Charcot-Marie-Tooth. Poisson z&egrave;bre montrent un excellent mod&egrave;le animal de nombreux troubles neurologiques, et ce syst&egrave;me a la possibilit&eacute; d&#39;informer notre compr&eacute;hension des maladies li&eacute;es au DNM2. Actuellement, on conna&icirc;t les orthologues chez le poisson z&egrave;bre endog&egrave;ne &agrave; la DNM2 humaine. Dans cette &eacute;tude, nous caract&eacute;risons deux poisson-z&egrave;bre dynamine-2 g&egrave;nes, dnm2 et dnm2-like. Les deux orthologues sont structurellement semblables aux DNM2 humaine au niveau g&eacute;n&eacute;tique et prot&eacute;ines. Elles sont exprim&eacute;es au cours du d&eacute;veloppement pr&eacute;coce et dans tous les tissus adultes. Pr&eacute;cipitation de dnm2 et des produits de g&egrave;ne de type dnm2 a entra&icirc;n&eacute; des anomalies morphologiques au cours du d&eacute;veloppement, et expression de l&#39;ARN de DNM2 humaine a sauv&eacute; ces ph&eacute;notypes. Nos r&eacute;sultats sugg&egrave;rent que dnm2 et dnm2-like sont orthologues de DNM2 humaine, et qu&#39;ils sont requis pour le d&eacute;veloppement du poisson z&egrave;bre normal.

Deux g&egrave;nes de la dynamine-2 sont n&eacute;cessaires pour le d&eacute;veloppement du poisson z&egrave;bre normal.

Dynamin-2 (DNM2) &egrave; un grande GTPase coinvolti in endocytosis clathrin-mediata e relativi percorsi di traffico. Le mutazioni umane DNM2 causano due distinti disturbi neuromuscolari: centronucleare miopatia e malattia di Charcot-Marie-Tooth. Zebrafish hanno dimostrato di essere un eccellente modello animale per molti disturbi di natura neurologica, e questo sistema ha il potenziale per informare la nostra comprensione della malattia DNM2-correlati. Attualmente, poco si sa circa la orthologs di zebrafish endogeno a DNM2 umana. In questo studio, abbiamo caratterizzano due zebrafish dynamin-2 geni, dnm2 e dnm2-come. Entrambi orthologs sono strutturalmente simili a DNM2 umana, a livello del gene e della proteina. Essi sono espressi durante lo sviluppo precoce e in tutti i tessuti adulti esaminati. Atterramento di dnm2 e prodotti simili dnm2 gene ha provocato estese anomalie morfologiche durante lo sviluppo, ed espressione di umana DNM2 RNA salvato questi fenotipi. I nostri risultati suggeriscono che dnm2 e dnm2-come sono orthologs a DNM2 umana, e che sono necessari per lo sviluppo normale di zebrafish.

Due dynamin-2 geni sono necessari per lo sviluppo normale di zebrafish.

DNM2 is a ubiquitously expressed GTPase that regulates multiple subcellular processes. Mutations in DNM2 are a common cause of centronuclear myopathy, a severe disorder characterized by altered skeletal muscle structure and function. The precise mechanisms underlying disease-associated DNM2 mutations are unresolved. We examined the common DNM2-S619L mutation using both in vitro and in vivo approaches. Expression of DNM2-S619L in zebrafish led to accumulation of aberrant vesicular structures and to defective excitation-contraction coupling. Expression of DNM2-S619L in COS7 cells resulted in defective BIN1-dependent tubule formation. These data suggest that DNM2-S619L may cause disease, in part, by interfering with membrane tubulation.

Defective tubulation associated with the myopathy causing S619L DNM2 mutation.

Duchenne muscular dystrophy (DMD) is a common and relentlessly progressive muscle disease. Some interventions have been identified that modestly slow progression and prolong survival, but more meaningful therapies are lacking. The goal of this study is to identify new therapeutic pathways for DMD using a zebrafish model of the disease. To accomplish this, we performed a non-biased drug screen in sapje, a zebrafish line with a recessive nonsense mutation in dystrophin. We identified 6 positive hits (out of 640 total drugs tested) by their ability to prevent abnormal birefringence in sapje. Follow-up analyses demonstrated that fluoxetine, a selective serotonin reuptake inhibitor (SSRI), provided the most substantial benefit. Morpholino-based experimentation confirmed that modulation of the serotonin pathway alone can prevent the dystrophic phenotype, and transcriptomic analysis revealed changes in calcium homeostasis as a potential mechanism. In all, we demonstrate that monoamine agonists can prevent disease in a vertebrate model of DMD. Given the safe and widespread use of SSRIs in clinical practice, our study identifies an attractive target pathway for therapy development.

Fluoxetine prevents dystrophic changes in a zebrafish model of Duchenne muscular dystrophy.

Nemaline myopathy (NM) is a genetic muscle disorder characterized by muscle dysfunction and electron-dense protein accumulations (nemaline bodies) in myofibers. Pathogenic mutations have been described in 9 genes to date, but the genetic basis remains unknown in many cases. Here, using an approach that combined whole-exome sequencing (WES) and Sanger sequencing, we identified homozygous or compound heterozygous variants in LMOD3 in 21 patients from 14 families with severe, usually lethal, NM. LMOD3 encodes leiomodin-3 (LMOD3), a 65-kDa protein expressed in skeletal and cardiac muscle. LMOD3 was expressed from early stages of muscle differentiation; localized to actin thin filaments, with enrichment near the pointed ends; and had strong actin filament-nucleating activity. Loss of LMOD3 in patient muscle resulted in shortening and disorganization of thin filaments. Knockdown of lmod3 in zebrafish replicated NM-associated functional and pathological phenotypes. Together, these findings indicate that mutations in the gene encoding LMOD3 underlie congenital myopathy and demonstrate that LMOD3 is essential for the organization of sarcomeric thin filaments in skeletal muscle.

Ann E. Davidson

Departments of Pediatrics and Neurology

University of Michigan

Analysis of Embryonic and Larval Zebrafish Skeletal Myofibers from Dissociated Preparations

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

Ann E. Davidson

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Departments of Pediatrics and Neurology

University of Michigan

Analysis of Embryonic and Larval Zebrafish Skeletal Myofibers from Dissociated Preparations

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

Departments of Pediatrics and Neurology