Increased expression of acetylcholine receptors in the diaphragm muscle of MDX mice.
The absence of dystrophin in Duchenne muscular dystrophy (DMD) and in the mutant mdx mouse causes muscle degeneration and disruption of the neuromuscular junction. Based on evidence from the denervation-like properties of these muscles, we assessed the ligand-binding constants of nicotinic acetylcholine receptors (nAChRs) and the mRNA expression of individual subunits in membrane preparations of diaphragm muscles from adult (4-month-old) and aged (20-month-old) control and mdx mice. The concentration of nAChRs as determined by the maximal specific [(125)I]-alpha-bungarotoxin binding (Bmax) in the muscle membranes did not change with aging in both animal strains. When compared to age-matched control groups, the Bmax in mdx muscles was increased by 65% in adults, and by 103% in aged mice with no alteration of toxin affinity for nAChRs. Reverse-transcription polymerase chain reaction assays showed that mRNA transcripts for the nAChR alpha1, gamma, alpha7, and beta2, but not the epsilon subunits, were more abundant in mdx than in control muscles. The results indicate increased expression of extrajunctional nAChRs in the mdx diaphragm and reflect impairment of nAChR regulation in dystrophin-deficient muscles. These observations may be related to the resistance to nondepolarizing muscle relaxants and the high sensitivity to depolarizing agents reported in DMD patients.
Ghedini
Paulo C
PC
Department of Pharmacology, Natural Products Section, Universidade Federal de São Paulo, 04044-020, Rua Três de Maio 100, São Paulo, SP, Brazil.
Viel
Tânia A
TA
Honda
Luciana
L
Avellar
Maria Christina W
MC
Godinho
Rosely O
RO
Lima-Landman
Maria Teresa R
MT
Lapa
Antônio J
AJ
Souccar
Caden
C
D000818:Animals; D002038:Bungarotoxins; D003627:Data Interpretation, Statistical; D003964:Diaphragm / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D011950:Receptors, Cholinergic / Q000096:biosynthesis* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002038:Bungarotoxins; D011950:Receptors, Cholinergic