The distribution and time course of brain-derived neurotrophic factor expression after plasmid DNA transfer to adult rat spinal cord

• Project/Area Number: 13671499
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: FUKUI MEDICAL UNIVERSITY
• Principal Investigator: UCHIDA Kenzo FUKUI MEDICAL UNIVERSITY, FACULTY OF MEDICINE, STUFF, 医学部, 助手 (60273009)
• Co-Investigator(Kenkyū-buntansha): BABA Hisatoshi FUKUI MEDICAL UNIVERSITY, FACULTY OF MEDICINE, PROFESSOR, 医学部, 教授 (00165060)
• Project Period (FY): 2001 – 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000); Fiscal Year 2001: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: gene transfer / liposome / brain-derived neurotrophic factor / cDNA / spinal cord injury / in vivo / rat

Research Abstract

We investigated the potential of non-viral vector-mediated brain-derived neurotrophic factor (BDNF) gene transfection in adult rat spinal cord. The potential of transfection of in vivo spinal cord was examined using three cationic lipids, Lipofectin^<TM>, LipofectAMINE^<TM> and LipofectAMINE^<TM> 2000, which have been proven to transfect genes into neuronal cells in vitro using plasmid DNA. After the direct injection of liposome cDNA complexes into thoracic spinal cord, histological analysis and immunoblot analysis demonstrated green fluorescent protein (GFP) and exogenous BDNF expression only in the group of LipofectAMINE^<TM> 2000/ cDNA complexes. In the group of LipofectAMINE^<TM> 2000/ cDNA complexes, fluorescent histochemical examination confirmed the presence of injected GFP around the injection areas for at least 5 days after injection. The transfected cells were identified reactive astrocytes, and some neuron. Both BDNF mRNA using RT-PCR and BDNF proteins using Western Blotting indicated a similar profile. The results suggest that cationic liposome-mediated delivery may be a practical method for gene transfer in spinal cord.

Research Products

• [Publications] Uchida, K., Baba, H., Furukawa, S., Omiya, M., Kokubo, Y., Kubota, C., Nakajima.: "Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy)"Acta neuropathologica. (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Uchida, K., Baba, H., Maezawa, Y., Kubota, C.: "Progressive changes of neurofilament proteins and growth-associated protein-43 immunoreactivities at the site of cervical spinal cord compression in spinal hyperostotic mice"Spine. 27. 480-486 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Yamaura, I., Yone, K., Nakahara, S., Nagamine, T., Baba, H., Uchida, K., Komiya, S.: "Mechanism of destructive pathologic changes in the spinal cord under chronic mechanical compression"Spine. 27. 21-26 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Uchida, K., Baba, H., Furukawa, S., Omiya, M., Kokubo, Y., Kubota, C., Nakajima.: "Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy)"Acta neuropathologica. 106-1. 29-36 (2003)
• [Publications] Uchida, K., Baba, H., Maezawa, Y., Kubota, C.: "Progressive changes of neurofilament proteins and growth-associated protein-43 immunoreactivities at the site of cervical spinal cord compression in spinal hyperostotic mice"Spine. 27. 480-486 (2002)
• [Publications] Yamaura, I., Yone, K., Nakahara, S., Nagamine, T Baba, H., Uchida, K., Komiya, S: "Mechanism of destructive pathologic changes in the spinal cord under chronic mechanical compression"Spine. 27. 21-26 (2002)