Presence of long and short dystrophin and/or utrophin products in <i>Torpedo marmorata</i> peripheral nerves

Rivier, François; Robert, Aude; Latouche, Jacqueline; Hugon, Gérald; Mornet, Dominique

doi:10.1016/0014-5793(95)01476-4

Cited by 9 publications

(11 citation statements)

References 34 publications

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“…However, the degree of homology is similar to that in the C-terminal region of dystrophin and utrophin, and immunization with sequences from these proteins has also resulted in specific and cross-reactive antibodies. 29,30 The chosen regions of the human calpain 3 sequence seem to share homology with a number of other proteins in different species, judging from the range of bands detected in different tissues and different species with these antibodies. In particular, the exon 8 immunogen seems to share some homology with myosin heavy chain, because both polyclonal antibodies and these monoclonal antibodies showed some immunoreactivity toward this protein.…”

Section: Characterization Of Epitopesmentioning

confidence: 99%

Characterization of Monoclonal Antibodies to Calpain 3 and Protein Expression in Muscle from Patients with Limb-Girdle Muscular Dystrophy Type 2A

Anderson

Davison

Moss

et al. 1998

The American Journal of Pathology

144

115

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Monoclonal antibodies were raised to two regions of calpain 3 (muscle-specific calcium-activated neutral protease), which is the product of the gene that is defective in limb-girdle muscular dystrophy type 2A. The antibodies produced characteristic patterns of bands on Western blots: normal calpain 3 protein was represented by bands at 94 kd, plus additional fragments at approximately 60 or 30 kd, according to the antibody used. Specificity was confirmed by the loss of all bands in patients with null gene mutations. The "normal" profile of bands was observed in muscle from 33 control subjects and 70 disease-control patients. Calpain 3 protein was found to be extremely stable in fresh human muscle, with full-size protein being detected 8 hours after the muscle had been removed. Blots of muscle from nine limb-girdle muscular dystrophy type 2A patients with defined mutations showed variation in protein expression, with seven showing a clear reduction in the abundance of protein detected. No simple relationship was found between the abundance and clinical severity. Two patients showed normal expression of the full-size 94 kd band accompanied by a clear reduction in the smaller fragments. This pattern was also observed in one patient with an undefined form of limb-girdle dystrophy. These results indicate that immunodiagnosis is feasible, but caution will need to be exercised with the interpretation of near-normal protein profiles.

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Section: Characterization Of Epitopesmentioning

confidence: 99%

Characterization of Monoclonal Antibodies to Calpain 3 and Protein Expression in Muscle from Patients with Limb-Girdle Muscular Dystrophy Type 2A

Anderson

Davison

Moss

et al. 1998

The American Journal of Pathology

144

115

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“…The specificity of each monoclonal antibody was previously confirmed in muscle and nerve tissues of rabbit (Fabbrizio et al, 1995), Torpedo (Rivier et al, 1996), and adult bovine heart (Rivier et al, 1999).We also produced new polyclonal antibodies directed against dystrophin, utrophin, α-dystrobrevin, epsilon-sarcoglycan and sarcospan that were obtained by injecting synthetic peptides corresponding to their C-terminal sequence (SRGNIPGKPMREDTM), (CPNVP-SRPQAM), (GVSYVPYCRS), (PQQQTTGKWYP) and (SLTAESEGPQQKI) as antigens, respectively, according to a previously described protocol (Rivier et al, 1996). In this study, we also used other polyclonal antibodies directed specifically against each sarcoglycan, as follows α (residues PLILDQH), β (residues AGYIPIDEDRHL), γ (residues VREQYT-TAEGI), δ (recombinant protein, residues 84-290), and β-dystroglycan (residues PPPYVPP).…”

Section: Antibodiesmentioning

confidence: 79%

“…We used a large battery of specific monoclonal and polyclonal antibodies produced in our laboratory. Their specificity was already described in previous studies related to dystrophin, utrophin and associated proteins Rivier et al, 1996;. We used Western blot technique to determine the presence and size of the molecules detected.Then, in a second step, we analyzed the histoimmuno-localisation of these molecules in various cell membranes.…”

mentioning

confidence: 99%

Dystrophin and dystrophin-associated protein in muscles and nerves from monkey

Royuela

Chazalette²,

Rivier³

et al. 2009

Eur J Histochem

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Since all organs (i.e. skeletal, cardiac, smooth muscles and sciatic nerve) are never only taken from a single patient, all these tissues were obtained from one cynomolgus monkey, a model closely resembling humans. This work describes an up-to-date reinvestigation of the dystrophin-glycoprotein complex and related molecules in various monkey tissues such those cited above. We used monoclonal and polyclonal antibodies produced in our laboratory, which are directed against dystrophin, utrophin, short-dystrophin products, α-dystrobrevin, β-dystroglycan, α-syntrophin, α-, β-, γ-, δ-, ε-sarcoglycan, and sarcospan. For each molecule, we determined their molecular weight and tissue localization. Regardless of the tissue analyzed, at least one dystrophin or utrophin as full-length molecule and one short-dystrophin product or dystrobrevin as proteins belonging to the dystrophin superfamily were found. β-dystroglycan, β and δ sarcoglycans were always detected, while other sarcoglycans varied from all to only three components. ε sarcoglycan appears to be specific to smooth muscle, which is devoid of α sarcoglycan. Sarcospan is only absent from sciatic nerve structures. Among the different muscles investigated in this study, short dystrophin products are only present in cardiac muscle. All of these findings are summarized in one table, which highlight in one single animal the variability of the dystrophin-glycoprotein complex components in relation with the organ studied. This statement is important because any attempt to estimate protein restoration needs in each study the knowledge of the expected components that should be considered normal.

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“…These domains are implicated in interactions with β-DG (Tinsley et al 1992). Like full-length utrophin, the Up71 isoform is ubiquitously expressed and was detected by our team and others in the peripheral nervous system and the small peripheral arteries (Fabbrizio et al 1995;Rivier et al 1996Rivier et al , 1997Wilson et al 1999). To date there has been little information about the expression of this short transcript and its implication in DMD pathology.…”

Section: Discussionmentioning

confidence: 99%

“…Monoclonal antibody (5A3) that recognized both dystrophin and utrophin was produced and characterized, as previously described (Fabbrizio et al 1995;Rivier et al 1996;Hernandez-Gonzalez et al 2005). Rabbit polyclonal antibody pan-Na + and the Schwann cell marker S-100 antibody were from Chemicon International.…”

Section: Antibodiesmentioning

confidence: 99%

Effect of β-dystroglycan processing on utrophin/Dp116 anchorage in normal and mdx mouse Schwann cell membrane

et al. 2006

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In the peripheral nervous system, utrophin and the short dystrophin isoform (Dp116) are co-localized at the outermost layer of the myelin sheath of nerve fibers; together with the dystroglycan complex. Dp116 is associated with multiple glycoproteins, i.e. sarcoglycans, and alpha- and beta-dystroglycan, which anchor the cytoplasmic protein subcomplex to the extracellular basal lamina. In peripheral nerve, matrix metalloproteinase activity disrupts the dystroglycan complex by cleaving the extracellular domain of beta-dystroglycan. Metalloproteinase creates a 30 kDa fragment of beta-dystroglycan, leading to a disruption of the link between the extracellular matrix and the cell membrane. Here we asked if the processing of the beta-dystroglycan could influence the anchorage of Dp116 and/or utrophin in normal and mdx Schwann cell membrane. We showed that metalloproteinase-9 was more activated in mdx nerve than in wild-type ones. This activation leads to an accumulation of the 30 kDa beta-dystroglycan isoform and has an impact on the anchorage of Dp116 and utrophin isoforms in mdx Schwann cells membrane. Our results showed that Dp116 had greater affinity to the full length form of beta-dystroglycan than the 30 kDa form. Moreover, we showed for the first time that the short isoform of utrophin (Up71) was over-expressed in mdx Schwann cells compared with wild-type. In addition, this utrophin isoform (Up71) seems to have greater affinity to the 30 kDa beta-dystroglycan which could explain the increased stabilization of this 30 kDa form at the membrane compartment. Our results highlight the potential participation of the short utrophin isoform and the cleaved form of beta-dystroglycan in mdx Schwann cell membrane architecture. We proposed that these two proteins could be implicated in Schwann cell proliferation in response to a microenvironment stress such as mediated by accumulating macrophages in mdx mouse muscle inflammation sites.

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Presence of long and short dystrophin and/or utrophin products in Torpedo marmorata peripheral nerves

Cited by 9 publications

References 34 publications

Characterization of Monoclonal Antibodies to Calpain 3 and Protein Expression in Muscle from Patients with Limb-Girdle Muscular Dystrophy Type 2A

Characterization of Monoclonal Antibodies to Calpain 3 and Protein Expression in Muscle from Patients with Limb-Girdle Muscular Dystrophy Type 2A

Dystrophin and dystrophin-associated protein in muscles and nerves from monkey

Effect of β-dystroglycan processing on utrophin/Dp116 anchorage in normal and mdx mouse Schwann cell membrane

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