Nestin-GFP Transgene Reveals Neural Precursor Cells in Adult Skeletal Muscle

Background: Therapy for neural lesions or degenerative diseases relies mainly on finding transplantable active precursor cells. Identifying them in peripheral tissues accessible for biopsy, outside the central nervous system, would circumvent the serious immunological and ethical concerns impeding cell therapy. Methodology/Principal Findings: In this study, we isolated neural progenitor cells in cultured adult skeletal muscle from transgenic mice in which nestin regulatory elements control GFP expression. These cells also expressed the early neural marker Tuj1 and light and heavy neurofilament but not S100b, indicating that they express typical neural but not Schwann cell markers. GFP+/Tuj1+ cells were also negative for the endothelial and pericyte markers CD31 and a-smooth muscle actin, respectively. We established their a) functional response to glutamate in patch-clamp recordings; b) interstitial mesenchymal origin; c) replicative capacity; and d) the environment necessary for their survival after fluorescenceactivated cell sorting. Conclusions/Significance: We propose that the decline in nestin-GFP expression in muscle progenitor cells and its persistence in neural precursor cells in muscle cultures provide an invaluable tool for isolating a population of predifferentiated neural cells with therapeutic potential

O ultrassom terapêutico na medula espinhal acelera a regeneração do nervo ciático de ratos Therapeutic ultrasound on the spinal cord accelerates regeneration of the sciatic nerve in rats

OBJETIVO: Estudar os efeitos da irradiação ultrassônica de baixa intensidade aplicada sobre a medula espinhal na regeneração do nervo ciático de ratos após lesão por esmagamento controlado, avaliando os resultados pelo índice funcional do ciático (SFI), medido nas imagens vídeo-filmadas das plantas das patas. MÉTODOS: Dezoito ratos foram submetidos a esmagamento controlado (do nervo ciático direito e divididos em dois grupos de acordo com o tratamento: Grupo 1 (n=9), irradiação simulada; Grupo 2 (n=9), irradiação efetiva. Irradiação ultrassônica de baixa intensidade foi iniciada no 7º dia pós-operatório e aplicada diariamente por 6 semanas. Imagens das plantas das patas dos animais foram vídeo-filmadas em uma esteira transparente sob velocidade controlada a intervalos semanais até a 6ª semana de irradiação e o correspondente SFI medido com um programa de computador específico. RESULTADOS: O SFI durante a 1ª e a 6ª semana de tratamento foi de -59,12 e -12,55 no Grupo 1, e -53,31 e -1,32 no Grupo 2, indicando uma melhora de 79% e 97%, respectivamente, mas as diferenças entre os grupos somente foram significantes (p<0,05) durante a 3ª semana de tratamento. CONCLUSÃO: Os autores concluem que o ultrassom terapêutico de baixa intensidade estimula a regeneração nervosa, com significância durante a 3ª semana de tratamento. Nivel de Evidência II, Prospectivo Comparativo.<br>OBJECTIVE: To study the effects of low intensity ultrasound irradiation applied on the spinal cord on the regeneration of the rat's sciatic nerve after a controlled crush injury, evaluating the functional results of the sciatic functional index as measured on the video recorded images of the foot sole. METHODS: Eighteen rats were submitted to a controlled crush injury of the right sciatic nerve and divided into two groups according to the treatment: Group 1 (n=9), simulated irradiation; Group 2 (n=9), effective irradiation. Low-intensity ultrasound irradiation was started on the 7th postoperative day and applied daily for 6 weeks. Images of the animals´ foot sole were video recorded on a see-through treadmill type walking belt machine at weekly intervals until the 6th week of irradiation and the corresponding sciatic functional index (SFI) was measured with specific software. RESULTS: The SFI during the first and last week of treatment was -59.12 and -12.55 in Group 1, -53.31 and -1.32 in Group 2, indicating a 79% and 97% improvement, respectively, but differences between groups were only significant (p<0.05) during the third week. CONCLUSION: The authors conclude that low intensity therapeutic ultrasound enhances nerve regeneration, with significance during the 3rd week of treatment. Level of Evidence: Level II, prospective comparative study