کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
6269593 1295148 2011 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Adeno-associated viral vector-mediated gene transduction in mesencephalic slice culture
موضوعات مرتبط
علوم زیستی و بیوفناوری علم عصب شناسی علوم اعصاب (عمومی)
پیش نمایش صفحه اول مقاله
Adeno-associated viral vector-mediated gene transduction in mesencephalic slice culture
چکیده انگلیسی

Adeno-associated viral (AAV) vector is a non-pathogenic vehicle that is suitable for the delivery of foreign genes into non-dividing neuronal cells. This vector has been utilized for in vivo neurological research and in clinical trials of gene therapy for neurodegenerative disorders. Viral vector-mediated gene delivery has the limitation that progressive changes in cellular phenotype cannot be monitored in living animals. To visualize living neurons transduced with foreign genes in vitro, we used cultured mesencephalic tissue harboring living dopaminergic (DA) neurons and examined cellular tropism of serotype-1 and serotype-2 AAV vectors in a culture system. The viability of DA neurons was evaluated using transgenic mice carrying enhanced green fluorescent protein under the control of the rat tyrosine hydroxylase (TH) promoter, which enables the visualization of living DA cells in the substantia nigra. Apoptosis of a subset of neuronal cells was noted within one day of culture. After 7 days, the serotype-1 AAV vector had successfully delivered the foreign gene into neurons and astrocytes, and serotype-2 AAV vector was able to transduce TH-positive DA neurons efficiently. Our method should be useful for in vitro investigations of pathological changes in DA neurons following transduction with foreign genes.

► Mesencephalic tissue culture harboring living dopaminergic neurons was generated. ► Cultured tissues were transduced with recombinant adeno-associated viral (rAAV) vector. ► Cellular tropism of serotype-1 and 2 rAAV vectors was investigated in the culture system.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Neuroscience Methods - Volume 201, Issue 1, 30 September 2011, Pages 55-60
نویسندگان
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