Li Jianhong, Chian Ru-Ju, Ay Ilknur, Kashi Brenda B, Celia Samuel A, Tamrazian Eric, Pepinsky R Blake, Fishman Paul S, Brown Robert H, Francis Jonathan W
Cecil B Day Laboratory for Neuromuscular Research, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA.
Biochem Biophys Res Commun. 2009 Dec 18;390(3):947-51. doi: 10.1016/j.bbrc.2009.10.083. Epub 2009 Oct 21.
With a view toward improving delivery of exogenous glial cell line-derived neurotrophic factor (GDNF) to CNS motor neurons in vivo, we evaluated the bioavailability and pharmacological activity of a recombinant GDNF:tetanus toxin C-fragment fusion protein in mouse CNS. Following intramuscular injection, GDNF:TTC but not recombinant GDNF (rGDNF) produced strong GDNF immunostaining within ventral horn cells of the spinal cord. Intrathecal infusion of GDNF:TTC resulted in tissue concentrations of GDNF in lumbar spinal cord that were at least 150-fold higher than those in mice treated with rGDNF. While levels of immunoreactive choline acetyltransferase and GFRalpha-1 in lumbar cord were not altered significantly by intrathecal infusion of rGNDF, GDNF:TTC, or TTC, only rGDNF and GDNF:TTC caused significant weight loss following intracerebroventricular infusion. These studies indicate that insect cell-derived GDNF:TTC retains its bi-functional activity in mammalian CNS in vivo and improves delivery of GDNF to spinal cord following intramuscular- or intrathecal administration.
为了提高向体内中枢神经系统运动神经元递送外源性胶质细胞源性神经营养因子（GDNF）的效率，我们评估了重组GDNF：破伤风毒素C片段融合蛋白在小鼠中枢神经系统中的生物利用度和药理活性。肌肉注射后，GDNF：TTC而非重组GDNF（rGDNF）在脊髓腹角细胞内产生强烈的GDNF免疫染色。鞘内注射GDNF：TTC导致腰段脊髓中GDNF的组织浓度比用rGDNF处理的小鼠至少高150倍。虽然鞘内注射rGNDF、GDNF：TTC或TTC对腰段脊髓中免疫反应性胆碱乙酰转移酶和GFRalpha-1的水平没有显著改变，但只有rGDNF和GDNF：TTC在脑室内注射后导致显著体重减轻。这些研究表明，昆虫细胞来源的GDNF：TTC在哺乳动物中枢神经系统体内保留其双功能活性，并在肌肉注射或鞘内给药后改善GDNF向脊髓的递送。
