[Gene hACR-1 suppresses apoptosis of striated muscle fibres of m. quadriceps femoris after ballistic transfection of mdx mice].
Human minidystrophin gene (pSG5dys plasmid) and hACR-1 gene (pRc-CMV-10.1 plasmid) were cotransfected by means of "gene-gun" to M. quadriceps femoris of mdx mice. Effects of transfection on dystrophin expression and survival of striated muscle fibres (SMF) were studied on the 21st day after shots. In the control mdx dystrophin-positive muscular fibers [D(+)] SMF and destroyed SMF made 2.1 +/- 0.1 and 2.1 +/- 0.3%, respectively. In mice transfected with pSG5dys plasmid (20 mkg of DNA per mouse), the shares of D(+) SMF and dead SMF raised, respectively, up to 5.6 +/- 1.4 and 4.5 +/- 0.9%. Transfection of mice with pRc-CMV-10.1 (DNA dose is 20 mkg per mouse) reduced the levels of apoptosis in SMF and D(+) SMF level to 1.6 +/- 0.6 and 1.1 +/- 0.4%, respectively. Cotransfection by pSG5dys and pRc-CMV-10.1 plasmids (10 and 10 mkg of each plasmids DNA per mouse) reduced the share of D(+) SMF to 1.1 +/- 0.5% and SMF destruction to 0.9 +/- 0.3%. pSG5dys transfection considerably reduced the share of SMF having peripherally located nuclei, thus indicating a decrease in SMF differentiation level after transfection. Cotransfection of ACR-1 gene and a dystrophin minigene did not suppress further cytodifferentiation of mdx muscle fibers. A conclusion is made that ballistic transfection by hACR-1 gene reduces the level of apoptosis in mdx mice SMF without changing the level of SMF differentiation. The cotransfection of mdx mice muscle by hACR-1 and human minidystrophin gene reduces SMF destruction and supports SMF differentiation, too.
Mikhaĭlov
V M
VM
Institute of Cytology RAS, St. Petersburg. vmikhailov@mail.cytspb.rssi.ru
Kropotov
A V
AV
Tomilin
N V
NV
Zelenin
A V
AV
Krutilina
R I
RI
Kolesnikov
V A
VA
Ostapenko
O V
OV
Zelenina
I A
IA
Baranov
A N
AN
Shteĭn
G I
GI
Baranov
V S
VS
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D019470:Biolistics; D002454:Cell Differentiation / Q000187:drug effects; D016189:Dystrophin / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008297:Male; D051379:Mice; D018101:Mice, Inbred mdx; D008822:Mice, Transgenic; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000473:pathology*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000473:pathology*; D010957:Plasmids; D013848:Thigh; D014162:Transfection
D016189:Dystrophin