Adenoviral vector-mediated gene transfer and neurotransplantation: possibilities and limitations in grafting of the fetal rat suprachiasmatic nucleus

TY - JOUR

T1 - Adenoviral vector-mediated gene transfer and neurotransplantation

T2 - possibilities and limitations in grafting of the fetal rat suprachiasmatic nucleus

AU - van Esseveldt, K E

AU - Liu, R.

AU - Hermens, W.T.J.M.C.

AU - Verhaagen, J

AU - Boer, G J

PY - 1997/1

Y1 - 1997/1

N2 - Several studies have reported on the use of primary neural cells transduced by adenoviral vectors as donor cells in neurotransplantation. In the present investigation, we examined whether adenoviral vector-mediated gene transfer could be used to introduce and express a foreign gene in solid neural transplants of fetal suprachiasmatic nucleus (SCN) tissue. A recombinant adenoviral vector containing the reporter gene LacZ encoding for beta-galactosidase (Ad-LacZ) was used in order to establish the optimal procedure for ex vivo gene transfer. Expression of beta-galactosidase was dependent on the duration of the infection and on the vector concentration. Infection for a short period (< 4 h) with a high concentration of Ad-LacZ (3.4 x 10(9) pfu/ml), or for 18 h with a lower vector concentration (2 x 10(8) pfu/ml), resulted in expression of beta-galactosidase in a large number of neurons and glial cells up to 21 days in vitro. When infected fetal SCN tissue was implanted in the third ventricle of adult Wistar rats, expression was high after 8 days. After 21 days, the number of beta-galactosidase expressing cells had clearly declined, but expression remained present for at least 70 days. The method described in this paper might be applicable to introduce trophic factor genes in SCN grafts in order to support graft survival and to stimulate neurite outgrowth.

AB - Several studies have reported on the use of primary neural cells transduced by adenoviral vectors as donor cells in neurotransplantation. In the present investigation, we examined whether adenoviral vector-mediated gene transfer could be used to introduce and express a foreign gene in solid neural transplants of fetal suprachiasmatic nucleus (SCN) tissue. A recombinant adenoviral vector containing the reporter gene LacZ encoding for beta-galactosidase (Ad-LacZ) was used in order to establish the optimal procedure for ex vivo gene transfer. Expression of beta-galactosidase was dependent on the duration of the infection and on the vector concentration. Infection for a short period (< 4 h) with a high concentration of Ad-LacZ (3.4 x 10(9) pfu/ml), or for 18 h with a lower vector concentration (2 x 10(8) pfu/ml), resulted in expression of beta-galactosidase in a large number of neurons and glial cells up to 21 days in vitro. When infected fetal SCN tissue was implanted in the third ventricle of adult Wistar rats, expression was high after 8 days. After 21 days, the number of beta-galactosidase expressing cells had clearly declined, but expression remained present for at least 70 days. The method described in this paper might be applicable to introduce trophic factor genes in SCN grafts in order to support graft survival and to stimulate neurite outgrowth.

KW - Adenoviridae

KW - Animals

KW - Brain Tissue Transplantation

KW - Defective Viruses

KW - Fetal Tissue Transplantation

KW - Gene Expression Regulation, Developmental

KW - Genes, Reporter

KW - Genetic Therapy

KW - Genetic Vectors

KW - Parkinson Disease

KW - Rats

KW - Rats, Wistar

KW - Recombinant Fusion Proteins

KW - Suprachiasmatic Nucleus

KW - Transfection

KW - beta-Galactosidase

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

M3 - Article

C2 - 9125380

SN - 0165-0270

VL - 71

SP - 113

EP - 123

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

IS - 1

ER -