1 Institut National de la Santé et de la Recherche Médicale Unité Mixte de Recherche (UMR) 533, Faculté de Médecine, 44093 Nantes; 2 Centre National à la Recherche Scientifique UMR 7637, Ecole Supérieure de Physique et de Chimie Industrielle, 75005 Paris; and 3 Centre National à la Recherche Scientifique UMR 6548, 06108 Nice, France
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The mdx mouse is a model for human Duchenne muscular dystrophy (DMD), an X-linked degenerative disease of skeletal muscle tissue characterized by the absence of the dystrophin protein. The mdx mice display a much milder phenotype than DMD patients. After the first week of life when all mdx muscles evolve like muscles of young DMD patients, mdx hindlimb muscles substantially compensate for the lack of dystrophin, whereas mdx diaphragm muscle becomes progressively affected by the disease. We used cDNA microarrays to compare the expression profile of 1,082 genes, previously selected by a subtractive method, in control and mdx hindlimb and diaphragm muscles at 12 time points over the first year of the mouse life. We determined that 1) the dystrophin gene defect induced marked expression remodeling of 112 genes encoding proteins implicated in diverse muscle cell functions and 2) two-thirds of the observed transcriptomal anomalies differed between adult mdx hindlimb and diaphragm muscles. Our results showed that neither mdx diaphram muscle nor mdx hindlimb muscles evolve entirely like the human DMD muscles. This finding should be taken under consideration for the interpretation of future experiments using mdx mice as a model for therapeutic assays.
muscular dystrophy; mdx mice; complementary deoxyribonucleic acid microarray; differential gene expression
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
DYSTROPHIN IS A CYTOSKELETAL protein located at the inner face of the cell membrane and is associated with cytosolic membrane glycoproteins (58). Its primary role is to stabilize the muscle plasma membrane (6). Duchenne muscular dystrophy (DMD) is a degenerative skeletal muscle disease occurring in humans that is caused by various mutations or deletions in the dystrophin gene, located on the short arm of the X chromosome (Xp21; see Refs. 23 and 25). This X-linked recessive myopathy affects 1 in 3,500 boys and is characterized by an absence of dystrophin protein expression. Dystrophin deficiency induces a fast, progressive, and severe dystrophy of skeletal and heart muscles, leading to death around the age of 20 yr (18).
Mice of the strain C57BL/10ScSn-Dmdmdx (mdx; X-linked muscular dystrophy) are commonly used as a model for DMD (7). Because of a point-mutation at position 3185 of the murine dystrophin gene (23, 41), mdx muscles lack the subsarcolemmal dystrophin protein, similar to human DMD muscles. However, in mdx mice, the hindlimb muscles differ histologically from diaphragm muscle, which more closely resembles DMD muscles. Hindlimb muscles display an initial period of intense necrosis in 2- to 3-wk-old mice (24). Later on, a sustained regenerative activity allows mdx hindlimb muscles to compensate for the necrosis (8, 12). Despite the lack of dystrophin, mdx hindlimb muscles either do not show obvious functional disability or express only mild weakness in adult mice. In contrast, mdx diaphragm muscle initially exhibits a regeneration process that does not compensate for massive degeneration and fails to restore muscle structure and function. This leads to significant extracellular matrix proliferation (fibrosis) associated with severe functional changes, including a reduction in maximal force production, elasticity, and twitch speed. This dramatic evolution in mdx diaphragm muscle appears similar to that observed in skeletal limb muscles of DMD patients (46). These differences in evolution between mdx and DMD muscle types lead to a questioning of the validity of mdx mice as a model in studies of DMD.
The recent development of cDNA microarrays provided a powerful
experimental approach to determine simultaneously the molecular profile
of several hundreds to several thousands of genes in defined physiological conditions (5, 14). It offered the
possibility to identify molecular or cellular pathways not previously
associated with the disease (26). Recently, Chen et al.
(10) compared the expression profile of muscle biopsies
from patients with DMD or -sarcoglycan deficiency using
oligonucleotide arrays. This global analysis of expression profiles
revealed that 275/6,000 genes encoding proteins involved in diverse
cellular functions like metabolism, immune response, cell growth, or
differentiation and signaling were differentially regulated all along
the progression of both human diseases. The authors demonstrated a
multigenic impact of the unique genetic defect of dystrophin in DMD
muscles. Presently, more than a decade after the discovery of the
dystrophin gene and its involvement in DMD, no global view is available
comparing the transcriptomal states of mdx diaphragm and
hindlimb muscles and their evolution with aging.
In the present study, we used cDNA microarray technology to determine comparative gene expression profiles between hindlimb and diaphragm muscles of control (C57BL/10) and mdx mice. Previously, we had constructed four subtracted/normalized libraries containing cDNA clones of genes up- or downregulated in hindlimb or diaphragm muscles of 12-wk-old mdx mice (48) using a strategy based on suppression subtractive hybridization (SSH; see Ref. 13). These cDNA clones were printed on microarrays and profiled for gene expression in hindlimb and diaphragm muscles at 12 time points, ranging from 1 to 52 wk of age. We hypothesized that this global approach would provide a large-scale insight into the comparative remodeling of both mdx muscle types over the first year of the mouse life. Our aim was 1) to determine whether these murine muscle types actually represent a realistic animal model of the dramatic evolution observed in human DMD muscles and 2) to identify some of the transcriptomal landmarks and/or related molecular/cellular pathways leading to the distinct evolution between these two muscle types in mdx mice.
![]() |
MATERIALS AND METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Mouse strains and RNA isolation.
To determine which and how many genes were affected by the primary
deficiency of dystrophin during the life span of one year, the gene
expression profiles of mdx diaphragm and hindlimb muscles were compared with the control (C57BL/10) counterparts, using cDNA
microarrays. Breeding pairs of C57BL/10ScSn mice and of
C57BL/10ScSn-Dmdmdx/J mice (41)
were purchased from Jackson Laboratories and bred in a conventional
animal care facility (veterinarian school). Only male animals were
studied to avoid intersexual variation. Animals were killed by cervical
dislocation at 12 different ages (1, 2, 3, 4, 5, 7, 9, 12, 16, 24, 36,
and 52 wk) representative of the first year of the mouse life. Tissues
were rapidly dissected, snap-frozen in liquid nitrogen, and stored at
80°C as samples pooled from at least six animals of the same age to
eliminate any interindividual variation in expression levels. Total RNA was extracted using Trizol reagent (Life Technologies) and treated with
DNase I. Absence of genomic DNA was systematically assessed in parallel
PCR and RT-PCR experiments of purified RNA preparations using
actin-specific primers. Poly(A)+ RNA was isolated using the
Oligotex mRNA kit (Qiagen).
cDNA clone selection and functional classification.
The microarrays used in this study contained 1,082 different cDNA
clones; 1,019 out of 1,082 arrayed clones were issued from four
subtractive libraries previously prepared by SSH between control and
mdx hindlimb muscles and between control and mdx
diaphragm muscle of 12-wk-old mice (Table
1; see Refs. 13 and 48). The
sequence of 23% of these clones could not be matched to sequences deposited in public databases; therefore, they represented novel genes
(C. A. Dechesne and A. V. Tkatchenko, unpublished
observations). The remaining 77% represented known genes
encoding diverse protein components that were distributed evenly among
the six main functional categories (Table
2; see Ref. 1). Roughly
one-quarter of the clones is involved in metabolism. The large
representation of metabolic genes in our libraries correlates well with
the energy-demanding function of the mdx muscles that are
composed of numerous regenerated fibers. The next largest functional
category is that of cell signaling (containing 15% of the genes),
followed by gene/protein expression (11%). A few genes were
represented on the microarray by several clones containing sequences
from different parts of these genes. This redundancy provided useful
repetitive determinations and the possibility to average the expression
profiles of the same gene [e.g., parvalbumin, insulin-like growth
factor II (IGF-II), titin, etc.]. An additional population of 63 cDNA
clones was added to the microarrays. This population consisted of
3'-untranslated region PCR-amplified fragments from components of the
multiprotein dystrophin system, nuclear factor-B family proteins,
and a few potassium channels (Table 1). Information on all clones
(sequence, accession number, copy of the related publication; see Ref.
48) is available as Supplemental Material. Please refer to
the Supplemental Material1
for this article (published online at the American Journal of Physiology-Cell Physiology web site).
|
|
Microarray manufacturing, labeling, hybridizations, and imaging. cDNA microarrays were produced and hybridized as described previously (16, 17, 36). The 1,082 spotted cDNA fragments were cloned in the plasmid vector PCRII-TOPO (Invitrogen) and further amplified using universal sequences flanking the cloning site. The PCR products (0.4-1.5 kb) were precipitated, resuspended in 3× saline-sodium citrate, and robotically arrayed on polylysine-coated glass slides (GeneMachine; Omnigrid). First-strand complex cDNA probes were generated by incorporation of Cy3- or Cy5-dUTP (Amersham and Pharmacia) by reverse transcription of 1 µg of each pooled poly(A)+ RNA using Superscript II (Life Technologies). Each labeled cDNA from control and mdx muscles from mice of the 12 different ages was mixed with an equal amount of the corresponding 12-wk-old C57BL/10 mouse muscles used as control. To avoid unspecific DNA binding, mouse cot-1 DNA (GIBCO-BRL), yeast tRNA, and poly(dT) were added to the mixed Cy3- and Cy5-labeled probes before hybridization. Poly(A)+ RNA samples were alternatively labeled with either Cy3 or Cy5 to minimize the influence of either fluorochrome on the relative gene expression ratios. Hybridized arrays were scanned by fluorescence confocal microscopy (ScanArray 3000; GSI-Lumonics). Measurements were obtained separately for each fluorochrome at 10 µm/pixel resolution.
Analysis of array data.
Fluorescence values and ratios for each cDNA array were analyzed
using the ScanAlyze software package available at
http://rana.stanford.edu/software. Previously described
procedures and criteria were used to exclude defective array spots and
to correct for differences in DNA labeling efficiency
(16). All clones with either a weak hybridization signal
(<1,000) or a defective signal (dust, spot deformation, pixel
incoherence, etc.) were flagged and excluded from further analysis (see
Supplemental Material). All array experiments were (minimally)
performed as duplicates; duplicate expression ratios were averaged. Of
the arrays used for further analysis, at least 75% of the spots had a
regression correlation >0.85. In control experiments, the same
poly(A)+ RNA was divided into two parts, each independently
labeled with a different fluorochrome before mixing and hybridizing.
The median values of the rough differential expression ratios [(signal
Cy5)/(signal Cy3) for each gene] remained between 1.5 and 1.5 (Fig.
1A). This was consistent with
previous reports on microarray reproducibility and sensitivity
(16, 35-36, 38, 40, 45). A normalization procedure
was performed to compensate for moderate differences commonly observed
between Cy3 and Cy5 signals. Spots with a measured differential
expression ratio between 1.5 and 0.66 were selected in each microarray.
A linear regression was calculated for both fluorescent signals within
this clone subpopulation. The calculated coefficients were then applied
to all Cy5 values of the corresponding array to obtain normalized
differential expression ratios. In further experiments, performed using
different clone preparations, cDNA arrays, and cDNA targets, a
differential expression ratio of <0.5 or >2 was observed as the
threshold for experimentally reproducible differential expression
(Fig. 1B).
|
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Tissue and gene collection. To obtain a global and comparative vision of transcriptomal processes involved in control and mdx diaphragm and hindlimb muscles over the first year of mouse life, gene expression profiling was performed at 12 different time points. These time points encompassed two distinct mouse life periods that corresponded to young (1-5 wk old) and adult (7-52 wk old) age (29). Twelve-week-old control mice were initially chosen as the time point reference for the subtractive libraries (48) and were used in the present study as reference for all hybridization reactions. This age was generally considered pivotal for both mdx mice muscles (21, 29, 34, 37). It succeeds to the period of the most intense degeneration/regeneration processes affecting both types of mdx muscles (2-5 wk). It precedes the progressive degeneration and fibrosis that mainly affect adult diaphragm muscle (see additional Fig. 1 in Supplemental Material and Refs. 32 and 34).
The relevance of the 1,082 selected clones for the analysis of differential expression in mdx muscle tissue was enforced by the expression profiles of three different tissues of 12-wk-old control and mdx mice: diaphragm muscle, hindlimb muscles, and kidney. As shown in Fig. 2, a few tens of clones were found to be up- or downregulated in both muscle types, whereas the same clones were expressed equally in control and mdx kidney. These clones corresponded to clones previously identified as differentially expressed in mdx muscle using reverse and direct Northern blot analysis (13).
|
Collection of differentially expressed genes. A complete set of 48 expression ratios in hindlimb and diaphragm muscles from control and mdx mice at 12 different ages was obtained for 843/1,082 clones (Table 1). The remaining 239/1,082 clones were undetectable or only partially detectable. Some of the 843 fully informative clones showed persistent expression changes of >2.0 relative to control muscle at all 12 time points, whereas others were only differentially expressed at certain time points. A first classification consisted of the selection of clones that showed modulation of expression >2.0, in at least two successive time points. This initial classification of temporal gene expression ratios (16, 17, 36) revealed that, at early ages, numerous genes were up- or downregulated in both control and mdx muscles compared with the age of 12 wk. From the ages of 7-9 wk until 1 yr, most of the developmentally regulated genes maintained a persistent differential expression in mdx muscles but no significant expression variations in either control muscle.
The analytical SAM procedure (52) was applied to the collection of the 843 detected clones to identify those that presented statistically significant expression changes in mdx muscles. The SAM analysis was independently applied to comparative time-paired experiments over the (1-5 wk) young and (12-52 wk) adult periods of mouse life (29). On the basis of the degree of significance (52) and an applied global ratio difference of 2.0, SAM analysis produced lists of genes potentially deregulated by the dystrophin deficiency (see SAM-related table in Supplemental Material). Figure 3 summarizes the SAM results obtained from the time-paired comparisons between mdx and control hindlimb (Fig. 3A) and diaphragm (Fig. 3B) muscles, and between both mdx muscle types (Fig. 3C), during the adult life period. No significant gene expression differences were found between control hindlimb and diaphragm muscles in the adult period (Fig. 3D). This suggested that the expression differences detected in and between mdx muscles during the second part of the mouse life span were solely because of dystrophin deficiency. SAM analysis was also performed on the same four muscle pairs during the (1- to 5-wk) young period where numerous genes displayed large differential expression ratios ranging from 2 to 80. Some genes were found to be statistically different between control hindlimb and diaphragm muscles, as well as between control and mdx muscle types and between mdx hindlimb and diaphragm muscles. These observations showed the difficulty to distinguish between the genetic program of normal muscle development and its eventual perturbation because of the dystrophin defect.
|
|
Expression profiling in control and mdx muscles.
The 112 differential expression profiles were compared and classified
into four different groups, as shown in Fig.
5. Within each profile group, we
classified the cDNA clones by functional categories (Fig. 5). The first
group contained clones with temporal gene expression profiles similar
for both mdx muscle types but differing from profiles of
their control counterparts (n = 38, subset
I). In this group, dystrophin deficiency induced deregulation of
numerous genes, associated with membrane disorganization, sarcomere desintegrity, and gene expression. In both adult mdx muscle
types, the point mutation in the dystrophin gene clearly had similar multiple effects on diverse transcriptomal activities. The three remaining groups in Fig. 5 were composed of clones that showed differential expression, either only in mdx diaphragm muscle
(n = 21, subset IIa) or only in
mdx hindlimb muscles (n = 40, subset IIb), or that showed different differential expression in both types of mdx muscle (n = 13, subset
IIc).
|
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
This is the first report of large-scale temporal gene expression profiling in two mdx muscles. The data were obtained using arrays containing 1,082 cDNA clones selected from four substractive libraries prepared from control and mdx diaphragm and hindlimb muscles (48). The application of the stringent analytical method SAM (52) to the numerous expression data drastically limited the number of informative genes. Roughly 10% of all the clones previously isolated by SSH (48) were finally detected as being differentially expressed in our microarray experiments. Different factors [the redundancy observed in the SSH procedure applied on 12-wk-old muscles only (48), the cellular complexity of the tissue samples used for the targets, the insensitivity of the array method when analyzing weakly expressed genes, etc.] could explain the discrepancy observed between the two systems of gene selection and their limits of application.
The present differential gene set of 112 out of 1,082 in mdx
mice is parallel to the set of 275 out of 6,000 human DMD differential genes identified in a recent study analyzing expression profiles of
muscle biopsies of patients with DMD and patients with -sarcoglycan deficiency, using oligonucleotide arrays (see Table 3, Ref.
10, and
http://microarray.NMCTesearch.org/resources.htm). Both murine and human gene sets constitute the largest (but obviously not
exhaustive) collection of transcriptomal landmarks in
dystrophin-deficient muscles. A flow chart, indicating the pathological
cascades initiated by dystrophin deficiency, was proposed to interpret
the oligonucleotide microarray expression profiling study of human DMD
muscles (10). It listed the main molecular circuits or
pathways that are perturbed in dystrophic cells, their possible
progressive interactions, and the relationship between cell
autonomous and microenvironmental changes. With regard to the proposed
flow chart and the related profiles in human dystrophic muscles, two
main conclusions can be drawn from our expression results in
mdx muscles.
Conclusion 1: Dystrophin deficiency initiates globally identical pathological cascades in murine and human dystrophic muscles. The most central feature in all dystrophic muscles of both animal species clearly was the occurrence of perturbations in calcium homeostasis (21). This is obviously connected to the primary function of dystrophin, which is to stabilize the sarcolemmal membrane of myofibers. In DMD (4) and mdx myofibers (30, 51), dystrophin deficiency results in membrane lesions, leading to chronic leakage of intracellular constituents and increased intracellular calcium concentration (24). As previously documented in mdx hindlimb myofibers, this abnormal calcium accumulation causes most of the multiple progressive downstream consequences of the dystrophin defect by mediating an elevated rate of protein degradation (15, 44; Fig. 5, subset I).
In addition, numerous other (secondary) markers involved in proliferation/dedifferentiation and inflammation processes were identified in both murine and human dystrophic muscles, indicating that the absence of dystrophin initiates the same perturbations in the genetic machinery of both animal species. Examples were osteoblast-specific factor 2 (Osf2), IGF-II, lyzozyme M and P, etc., which were upregulated in DMD and both mdx muscles. Osf2, a chondrocyte-related transcript, was one of the strongest upregulated genes in both muscle types in adult mdx mice (expression ratios of 8 and 4 in diaphragm and hindlimb muscles, respectively). Its persistent upregulation, which was detected in both animal species (10), was linked to that of human versican, a large chondroitin sulfate proteoglycan known to stimulate the proliferation of chondrocytes, and to that of three other chondrocyte- and bone-related transcripts in dystrophic muscles (10, 56). Using the murine arrays, we identified novel markers not identified on the human array. Most of these markers belonged to the circuits of the proposed pathological flow (10). The central role of calcium homeostasis disruption in dystrophic cells was confirmed by expression variations in several genes: the parvalbumin gene, genes encoding proteins like calcium/calmodulin-dependent protein kinase, and a few protease inhibitors that are controlled by the intracellular calcium concentration. Our murine expression arrays revealed for the first time a concomitant and persistent upregulation of the transcriptionally related components H19 and IGF-II in mdx muscles. Normally, both genes are expressed in many tissues of endodermal and mesodermal origin during embryogenesis (28, 33), but they are downregulated at birth. IGF-II and H19 undergo genomic imprinting in control mouse and human tissues, resulting in parent-specific monoallelic expression. Perturbations in the expression of both genes, including loss of imprinting, were evidenced in several cancers, including rhabdomyosarcoma (9, 55). The persistent increased expression of both genes in mdx muscles (and of IGF-II in human dystrophic hindlimb muscles) underlines the importance of massive muscle cell proliferation and dedifferentiation in each mdx muscle type along the mouse life span. In parallel, it was recently documented that expression of IGF-II ameliorates the mdx phenotype by downregulating programmed cell death, as shown by a reduction of degenerative regions and of the number of centrally located nuclei (42, 43). In addition, a link between IGF-II expression and collagen accumulation was proposed recently, suggesting a role of this protein in the mechanism of fibrosis in dystrophic cells (22, 31). The question whether these factors really do have a beneficial influence in terms of potential therapeutic targets is still open. Our results on the expression of genes encoding dystrophin-related or -associated proteins in mdx muscles contrasted surprisingly with those obtained in DMD muscles (10). We found thatConclusion 2: Dystrophin deficiency causes different perturbations
in the transcriptomal circuits of mdx diaphragm and hindlimb muscles.
The observed expression differences (Fig. 5, subsets II)
could explain some of the known histological and functional differences between both mdx muscle types. Thus the opposite expression
levels of parvalbumin genes in both mdx muscle types
probably reflected differences in the intracellular calcium content.
Such differences may in turn modulate the calcium-dependent proteolysis
in a muscle type-specific way. Low levels of parvalbumin (<2.5-fold),
a protein that links calcium and a relaxing factor in fast muscles
(39), together with the concomitant downregulation of
genes encoding 1-protease inhibitors type 1, 3, and 5 (<2.2-fold), could provoke an intensed proteolytic activity within the
mdx diaphragm cells and a massive
degeneration/disorganization with aging. Relatively low levels of
calcium, resulting from higher contents of parvalbumins and of protease
inhibitors, apparently prevented the mdx hindlimb cells from
such a rapid and dramatic degeneration. These observations led us to
propose a more precise reexamination of the protective or deleterious
role of intracellular calcium levels and their influence on the
activity of related proteases in the dystrophic muscle cells. These
proteins could be used as either pharmaceutical or genetic targets for
future therapy.
![]() |
ACKNOWLEDGEMENTS |
---|
We thank Agnes Hivonnait, Patrick Guyot, and Jacques Noireaud for assistance with animal care and facilities. We are grateful to Denis Escande, Jean-Jacques Schott, and Jean Rossier for helpful discussions and reading of the manuscript and to Philippe Marc for initial assistance in interpreting microarray image data.
![]() |
FOOTNOTES |
---|
M. Steenman was supported by a grant from the Institut National de la Santé et de la Recherche Médicale. This work was supported by the Institut National de la Santé et de la Recherche Médicale, the Centre National à la Recherche Scientifique, l'Association Française contre les Myopathies, and la Fondation de la Recherche Médicale.
Address for reprint requests and other correspondence: J. J. Léger, Inserm U533, Faculté de Médecine, 1 rue Gaston Veil, 44093 Nantes, France (E-mail: Jean.Leger{at}nantes.inserm.fr).
1 Supplemental material to this article is available online at http://ajpcell.physiology.org/cgi/content/full/283/3/C773/DC1.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
April 24, 2002;10.1152/ajpcell.00112.2002
Received 12 March 2002; accepted in final form 17 April 2002.
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Adams, MD,
Kerlavage AR,
Fleischmann RD,
Fuldner RA,
Bult CI,
Lee NH,
Kirkness EF,
Weinstock KG,
Gocayne JD,
White O,
Initial assessment of human gene diversity and expression patterns based upon 83 million nucleotides of cDNA sequence.
Nature
377:
3-174,
1995[ISI][Medline].
2.
Barbiroli, B,
Funicello R,
Lotti S,
Montagna P,
Ferlini A,
and
Zaniol P.
31P-NMR spectroscopy of skeletal muscle in Becker dystrophy and DMD/BMD carriers. Altered rates of phosphate transport.
J Neurol Sci
109:
188-195,
1992[ISI][Medline].
3.
Beis, A,
Zammit VA,
and
Newsholme EA.
Activities of 3-hydroxybutyrate dehydrogenase, 3-oxoacid CoA-transferase and acetoacetyl-CoA thiolase in relation to ketone-body utilization in muscles from vertebrates and invertebrates.
Eur J Biochem
104:
209-215,
1980[Abstract].
4.
Bodensteiner, JB,
and
Engel AG.
Intracellular calcium accumulation in Duchenne muscular dystrophy and other myopathies: a study of 567,000 muscle fibers in 114 biopsies.
Neurology
28:
314-327,
1977.
5.
Brown, PO,
and
Botstein D.
Exploring the new world of the genome with DNA microarrays.
Nat Genet
21:
33-37,
1999[ISI][Medline].
6.
Brown, RH,
and
Hoffman EP.
Molecular biology of Duchenne muscular dystrophy.
Trends Neurol Sci
11:
480-483,
1988[ISI][Medline].
7.
Bulfield, G,
Siller WG,
Wight PAL,
and
Moore KJ.
X chromosome-linked muscular dystrophy (mdx) in the mouse.
Proc Natl Acad Sci USA
81:
1189-1192,
1984[Abstract].
8.
Carnwath, JW,
and
Shotton DM.
Muscular dystrophy in the mdx mouse: histopathology of the soleus and extensor digitorum longus muscles.
J Neurol Sci
80:
39-54,
1987[ISI][Medline].
9.
Casola, S,
Pedone PV,
Cavazzana AO,
Basso G,
Luksch R,
D'amore ES,
Carli M,
Bruni CB,
and
Riccio A.
Expression and parental imprinting of the H19 gene in human rhabdomyosarcoma.
Oncogene
14:
1503-1510,
1997[ISI][Medline].
10.
Chen, YW,
Zhao P,
Borup R,
and
Hoffman EP.
Expression profiling in the muscular dystrophies. Identification of novel aspects of molecular pathophysiology.
J Cell Biol
151:
1321-1336,
2000
11.
Chi, M,
Hintz MY,
Mc Kee CS,
Felder S,
Grant N,
Kaiser KK,
and
Lowry OH.
Effect of Duchenne muscular dystrophy on enzymes of energy metabolism in individual muscle fibers.
Metabolism
36:
761-767,
1987[ISI][Medline].
12.
Dangain, J,
and
Vrbova G.
Muscle development in mdx mutant mice.
Muscle Nerve
7:
700-704,
1984[ISI][Medline].
13.
Diatchenko, L,
Lau F,
Campbell AP,
Chenchik A,
Moqadam F,
Huang B,
Lukyanov S,
Lukyanov K,
Gurskaya N,
Sverdlov ED,
and
Siebert PD.
Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.
Proc Natl Acad Sci USA
93:
6025-6030,
1996
14.
Duggan, DJ,
Bittner M,
Chen Y,
Meltzer P,
and
Trent JM.
Expression profiling using cDNA microarrays.
Nat Genet
21:
10-14,
1999[ISI][Medline].
15.
Ebashi, S,
and
Sugita H.
The role of calcium in physiological and pathological processes of skeletal muscle.
In: Current Topics in Nerve and Muscle Research, edited by Aguayo AJ,
and Karpati G.. Amsterdam, The Netherlands: Excerpta Medica, 1979, p. 73-84.
16.
Eisen, MB,
and
Brown PO.
DNA arrays for analysis of gene expression.
Methods Enzymol
303:
179-205,
1999[ISI][Medline].
17.
Eisen, MB,
Spellman PT,
Brown PO,
and
Botstein D.
Cluster analysis and display of genome-wide expression patterns.
Proc Natl Acad Sci USA
95:
14863-14868,
1998
18.
Emery, AEH
Duchenne Muscular Dystrophy. Oxford Monographs on Medical Genetics (2nd ed.). Oxford, UK: Oxford University Press, 1993, vol. 24.
19.
Even, PC,
Decrouy A,
and
Chinet A.
defective regulation of energy metabolism in mdx-mouse skeletal muscles.
Biochem J
304:
649-654,
1994[ISI][Medline].
20.
Gannoun-Zaki, L,
Fournier-Bidoz S,
Le Cam G,
Chambon C,
Millasseau P,
Leger JJ,
and
Dechesne CA.
Down-regulation of mitochondrial mRNAs in the mdx mouse model for Duchenne muscular dystrophy.
FEBS Lett
375:
268-272,
1995[ISI][Medline].
21.
Gillis, JM.
Understanding dystrophinopathies: an inventory of the structural and functional consequences of the absence of dystrophin in muscles of the mdx mouse.
J Muscle Res Cell Motil
20:
605-625,
1999[ISI][Medline].
22.
Goldspink, G,
Fernandes K,
Williams PE,
and
Wells DJ.
Age-related changes in collagen gene expression in the muscles of mdx dystrophic and normal mice.
Neuromuscul Disord
4:
183-191,
1994[ISI][Medline].
23.
Hoffman, EP,
Brown RH,
and
Kunkel LM.
Dystrophin: the protein product of the Duchenne muscular dystrophy locus.
Cell
51:
919-928,
1987[ISI][Medline].
24.
Hoffman, EP,
and
Gorospe JR.
The animal models of Duchenne muscular dystrophy: windows on the pathophysiological consequences of dystrophin deficiency.
In: Ordering the Membrane Cytoskeleton Trilayer, edited by Mooseker MT,
and Morrow J.. New York: Academic, 1991, p. 113-154.
25.
Hoffman, EP,
and
Kunkel LM.
Dystrophin abnormalities in Duchenne/Becker muscular dystrophy.
Neuron
2:
1019-1029,
1989[ISI][Medline].
26.
Iyer, VR,
Eisen MB,
Ross DT,
Schuler G,
Moore T,
Lee JCF,
Trent JM,
Staudt LM,
Hudson J, Jr,
Boguski MS,
Lashkari D,
Shalon D,
Botstein D,
and
Brown PO.
The transciptional program in the response of human fibroblast to serum.
Science
283:
83-87,
1999
27.
Kuznetsov, AV,
Winkler K,
Wiedemann FR,
Von Bossanyi P,
Dietzmann K,
and
Kunz WS.
Impaired mitochondrial oxidative phosphorylation in skeletal muscle of the dystrophin-deficient mdx mouse.
Mol Cell Biochem
183:
87-96,
1998[ISI][Medline].
28.
Looijenga, LHJ,
Verkerk AJMH,
De Groot N,
Hochberg AA,
and
Oosterhuis JW.
H19 in normal development and neoplasia.
Mol Reprod Dev
46:
419-439,
1997[ISI][Medline].
29.
Lynch, GS,
Hinkle RT,
Chamberlain JS,
Brooks SV,
and
Faulkner JA.
Force and power output of fast and slow skeletal muscles from mdx mice 6-28 months old.
J Physiol
535:
591-600,
2001
30.
MacLennan, P,
McArdle A,
and
Edwards RHT
Effects of calcium on protein turnover of incubated muscles from mdx mice.
Am J Physiol Endocrinol Metab
260:
E594-E598,
1991
31.
Murphy, PG,
Loitz BJ,
Frank CB,
and
Hart DA.
Influence of exogenous growth factors on the synthesis and secretion of collagen types I and III by explants of normal and healing rabbit ligaments.
Biochem Cell Biol
72:
403-409,
1994[ISI][Medline].
32.
Niebroj-Dobosz, I,
Fidzianska A,
and
Glinka Z.
Comparative studies of Hindlimb and diaphragm muscles of mdx mice.
Basic Appl Myol
7:
381-386,
1997[ISI].
33.
Ohlsson, R,
and
Franklin G.
Normal development and neoplasia: the imprinting connection.
Int J Dev Biol
39:
869-876,
1995[ISI][Medline].
34.
Pastoret, C,
and
Sebille A.
mdx mice show progressive weakness and muscle deterioration with age.
J Neurol Sci
129:
97-105,
1995[ISI][Medline].
35.
Perou, CM,
Jeffrey SS,
Van De Rijn M,
Rees CA,
Eisen MB,
Ross DT,
Pergamenschikov A,
Williams CF,
Zhu SX,
Lee JC,
Lashkari D,
Shalon D,
Brown PO,
and
Botstein D.
Distinctive gene expression patterns in human mammary epithelial cells and breast cancers.
Proc Natl Acad Sci USA
96:
9212-9217,
1999
36.
Pollack, JR,
Perou CM,
Alizadeh AA,
Eisen MB,
Pergamenschikov A,
Williams CF,
Jeffrey SS,
Botstein D,
and
Brown PO.
Genome-wide analysis of DNA copy-number changes using cDNA microarrays.
Nat Genet
23:
41-46,
1999[ISI][Medline].
37.
Porter, JD,
Khanna S,
Kaminski HJ,
Rao JS,
Merriam AP,
Richmonds CR,
Leahy P,
Li J,
Guo W,
and
Andrade FH.
A chronic inflammatory response dominates the skeletal muscle molecular signature in dystrophin-deficient mdx mice.
Hum Mol Genet
11:
263-272,
2002
38.
Ross, DT,
Scherf U,
Eisen MB,
Perou CM,
Rees C,
Spellman P,
Lyer V,
Jeffrey SS,
Van Der Rijn M,
Waltham M,
Pergamenschikov A,
Lee JC,
Lashkari D,
Shalon D,
Myers TG,
Weinstein JN,
Botstein D,
and
Brown PO.
Systematic variation in gene expression patterns in human cancer cell lines.
Nat Genet
24:
227-235,
2000[ISI][Medline].
39.
Sano, M,
Yokota T,
Endo T,
and
Tsukagoshi H.
A developmental change in the content of parvalbumin in normal and dystrophic mouse (mdx) muscle.
J Neurol Sci
97:
261-272,
1990[ISI][Medline].
40.
Sehl, PD,
Tai JT,
Hillan KJ,
Brown LA,
Goddard A,
Yang R,
Jin H,
and
Lowe DG.
Application of cDNA microarrays in determining molecular phenotype in cardiac growth, development, and response to injury.
Circulation
101:
1990-1999,
2000
41.
Sicinski, P,
Geng Y,
Ryder-Cook AS,
Barnard MG,
Darlinson MG,
and
Barnard PJ.
The molecular basis of muscular dystrophy in the mdx mouse: a point mutation.
Science
244:
1578-1580,
1989[ISI][Medline].
42.
Smith, J,
Fowke G,
and
Schofield PN.
Programmed cell death in dystrophic (mdx) muscle is inhibited by IGF-II.
Cell Death Differ
2:
243-251,
1995[ISI].
43.
Smith, J,
Goldsmith C,
Ward A,
and
LeDieu R.
IGF-II ameliorates the dystrophic phenotype and coordinately down-regulates programmed cell death.
Cell Death Differ
11:
1109-1118,
2000.
44.
Spencer, M,
and
Tidball JG.
Calpain concentration is elevated although net calcium-dependent proteolysis is suppressed in dystrophin-deficient muscle.
Exp Cell Res
203:
107-114,
1992[ISI][Medline].
45.
Stanton, LW,
Garrard LJ,
Damm D,
Garrick BL,
Lam A,
Kapoun AM,
Zheng Q,
Protter AA,
Schreiner GF,
and
White RT.
Altered patterns of gene expression in response to myocardial infarction.
Circ Res
86:
939-945,
2000
46.
Stedman, HH,
Sweeney HL,
Shrager JB,
Maguire HC,
Panetieeri RA,
Petrof B,
Narusawa M,
Leferovich JM,
Sladky JT,
and
Kelly AM.
The mdx mouse diaphragm reproduces the degenerative changes of Duchenne muscular dystrophy.
Nature
352:
536-539,
1991[ISI][Medline].
47.
Tanabe, Y,
Esaki K,
and
Nomura T.
Skeletal muscle pathology in X chromosome-linked muscular dystrophy (mdx) mouse.
Acta Neuropathol (Berl)
69:
91-95,
1986[ISI][Medline].
48.
Tkatchenko, AV,
Le Cam G,
Leger JJ,
and
Dechesne CA.
Large-scale analysis of differential gene expression in the hindlimb muscles and diaphragm of mdx mouse.
Biochim Biophys Acta
1500:
17-30,
2000[ISI][Medline].
49.
Tkatchenko, AV,
Pietu G,
Cros N,
Gannoun-Zaki L,
Auffray C,
Leger JJ,
and
Dechesne CA.
Identification of altered gene expression in skeletal muscles from Duchenne muscular dystrophy patients.
Neuromuscul Disord
11:
269-277,
2001[ISI][Medline].
50.
Toyofuko, T,
Hoffman JR,
Zak R,
and
Carlson BM.
Expression of -cardiac and
-skeletal actin mRNAs in relation to innervation in regenerating and non-regenerating rat skeletal muscles.
Dev Dyn
193:
332-339,
1992[ISI][Medline].
51.
Turner, PR,
Westwood T,
Regen CM,
and
Steinhardt RA.
Increased protein degradation results from elevated free calcium levels found in muscle from mdx mice.
Nature
335:
735-738,
1988[ISI][Medline].
52.
Tusher, VG,
Tibshirani R,
and
Chu G.
Significance analysis of microarrays applied to the ionizing radiation response.
Proc Natl Acad Sci USA
98:
5116-5121,
2001
53.
Van Der Ven, PF,
Bartsch JW,
Gautel M,
Jockusch H,
and
Furst DO.
A functional knock-out of titin results in defective myofibrils assembly.
J Cell Sci
113:
1405-1414,
2000
54.
Whalen, RG,
Schwartz K,
Bouveret P,
Sell SM,
and
Gros F.
Contractile protein isoenzymes in muscle development: identification of an embryonic form of myosin heavy chain.
Proc Natl Acad Sci USA
76:
5197-5201,
1979[Abstract].
55.
Zhan, S,
Shapiro DN,
and
Helman LJ.
Activation of an imprinted allele of the insulin-like growth factor II gene implicated in rhabdomyosarcoma.
J Clin Invest
94:
445-448,
1994[ISI][Medline].
56.
Zhang, Y,
Cao L,
Kiani C,
Yang BL,
Hu W,
and
Yang BB.
Promotion of chondrocyte proliferation by versican mediated by G1 domain and EGF-like motifs.
J Cell Biochem
73:
445-457,
1999[ISI][Medline].
57.
Zisman, A,
Peroni OD,
Abel ED,
Michael MD,
Mauvais-Jarvis F,
Lowell BB,
Wojtaszewski JF,
Hirshman MF,
Virkamaki A,
Goodyear LJ,
Khan CR,
and
Khan BB.
Targeted disruption of the glucose transporter 4 selectively in muscle causes insulin resistance and glucose intolerance.
Nat Med
6:
924-928,
2000[ISI][Medline].
58.
Zubrzycka-Gaarn, EE,
Bulman DE,
Karpati G,
Burghes AHM,
Belfall B,
Klamut HJ,
Talbot J,
Hodges RS,
Ray PN,
and
Worton RG.
The Duchenne muscular dystrophy gene product is localized in sarcolemma of human skeletal muscle.
Nature
333:
466-469,
1988[ISI][Medline].