Structure of neuromuscular junctions and differentiation of striated muscle fibers in mdx mice after bone-marrow stem cell therapy

Sokolova, A. V.; Зенин, В. В.; Mikhailov, V. M.

doi:10.1134/s1990519x10030077

Cited by 5 publications

(7 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Local transplantation of Lin(-) BM cells in M. quadriceps femoris of mdx mice aggregates islets of bungarothoxin-positive substance for large NMJs but without branches. In this case the level of dystrophin synthesis did not exceed 2% [30]. The more extensive level of dystrophin synthesis in case of whole body irradiation 3 Gy followed by BM cells transplantation ( Table 2) correlates with reparation of NMJs and branches enrichment.…”

Section: Discussionmentioning

confidence: 68%

“…Study of NMJs structure and their resting potential was conducted for two reasons. We have shown early that local transplantation of BM stem cells to M. quadriceps femoris of unirradiated mdx mice partly restore the structure of NMJs [30]. To confirm the results we studied a structure of diaphragm NMJs on the transverse section of synapsis region of diaphragm SMFs by staning with Tetramethylrhodamine-αbungarotoxin after general transplantation of BM cells.…”

Section: Resultsmentioning

confidence: 82%

“…In case of local transplantation of BMSCs we did not observed the formation of branches. High concentration of islets and growth of square of NMJs taken place on the background of negligible dystrophin synthesis near 2% [30]. To estimate functional significance of the structural changes of diaphragm NMJs we studied the electrophysiological properties of NMJs through 4 months after BM transplantation (Table 4).…”

Section: Resultsmentioning

confidence: 99%

“…Immunohistochemistry 10 µm cryosections were cut with cryostat (Bright Co, Ltd, UK) after preliminary muscle freezing in liquid nitrogen. Dried sections were fixed in ethanol mixed with carbinol (1:1 v/v) for 1 min at -20°C or in 10% formalin for 30 minutes at room temperature [30].…”

Section: Methodsmentioning

confidence: 99%

“…The differences between groups were assessed using Student's t-test. The differences were considered to be significant at p < 0.05 [30].…”

Section: Nmjs Assessmentmentioning

confidence: 99%

See 4 more Smart Citations

Non-myeloablative Bone Marrow Stem Cell Transplantation for mdx Mice Myodystrophy Therapy

Mikhailov¹,

Sokolova²,

Кравцова³

et al. 2012

J Cell Sci Ther

View full text Add to dashboard Cite

muscle and participate in muscle repair [12,13] and in muscle of mdx mice after BM stem cells transplantation [6].Except blood and muscle cells BM stem cells are also involved in the cell differentiation and regeneration in lungs, liver, skin, gastrointestinal tract and of epithelium of thyroid gland [6,14,15].Irradiation is a mandatory preliminary step of BM stem cells Abstract Background: Mdx mice are experimental model of cureless human monogenic disease Duchenne Muscular Dystrophy (DMD). Hope for а cure is associated with the use of stem cells therapy particular but not exclusively. Analysis of multiple experimental results shows what intramuscularly transplantation of different types of cells of different origins with stem cells properties can't convert mutant striated muscles fibers (SMF) into wild type SMF. It was concluded that only replacement of mutant bone marrow (BM) by wild type BM cells can convert mutant SMF into SMF of wild type. Unfortunately X-ray irradiation of mdx mice at a lethal dose of 11, 7 or 5 Gy followed by transplantation of wild C57BL/6 mice BM cells did not increase SMF dystrophin synthesis. The aim of this study was to analyse a dystrophin synthesis by mdx mice striated muscles after x-ray irradiation with the dose of 3 Gy followed by C57BL/6 bone marrow cells transplantation. Also we investigate the reparation of structure of diaphragm muscle fibers NMJs. To confirm the functional significance of observed structural changes of NMJs an investigation of resting membrane potential of diaphragm muscle fiber NMJs was also conducted.Methods: 1-1.5 months old mdx mice were irradiated by x-ray at a dose of 3 Gy. Next day freshly prepared BM cells were injected intravenously in the amount of (15-20) x 10 6 cells per mouse. Animals were studied through 2, 4, and 6 months after transplantation. Each experimental group of mice included 3-8 animals. Mus. quadriceps femoris and diaphragm muscle fibers with their nerve-muscle junctions (NMJs) were under investigation.For chimerism registration a special study was conducted using transplantation of GFP-positive C57BL/6 BM cells to mdx mice after 3 Gy irradiation. Through 6 months BM cells were separated from long bones and smears were prepared. After carbinol fixation smears were stained by propidium iodide and studied on confocal microscope LSM 5 Pascal (Carl Zeiss, Germany) to count the part of GFP-positive cells in relation to whole quantity of nuclear cells. Results:We observed a stable growth of dystrophin synthesis after nonlethal X-ray irradiation at a dose of 3 Gy. The part of dystrophin positive SMF of M. quadriceps femoris increased from 1% up to 4% (2 months), 12% (4 months) and 27% (6 months) after transplantation. Growth of dystrophin synthesis was accompanied by the decrease of SMF death level, by increase of part of SMF without central nuclear up to 22%, by accumulation of MNJ branches and by reparation of resting membrane potentials. The part of GFP-positive cells between all cells with nuclear on the BM smears of chimeric GFP tran...

show abstract

Section: Discussionmentioning

confidence: 68%

Section: Resultsmentioning

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The differences between groups were assessed using Student's t-test. The differences were considered to be significant at p < 0.05 [30].…”

Section: Nmjs Assessmentmentioning

confidence: 99%