Recombinant GABAC receptors expressed in rat hippocampal neurons after infection with an adenovirus containing the human ρ1 subunit

N. Filippova, A. Sedelnikova, William Tyler, T. L. Whitworth, H. Fortinberry, D. S. Weiss

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

1. A recombinant adenovirus was generated with the human ρ1 GABAC receptor subunit (adeno-ρ). Patch-clamp and antibody staining were employed to confirm functional expression of recombinant ρ1 receptors after infection of human embryonic kidney cells (HEK293 cell line), human embryonic retinal cells (911 cell line), dissociated rat hippocampal neurons and cultured rat hippocampal slices. 2. Standard whole-cell recording and Western blot analysis using ρ1 GABAC receptor antibodies revealed that recombinant ρ1 receptors were expressed in HEK293 and 911 cells after adeno-ρ infection and exhibited properties similar to those of ρ1 receptors after standard transfection. 3. Cultured rat hippocampal neurons (postnatal day (P)3-P5) did not show a native GABAC-like current. After adeno-ρ infection, however, a GABAC-like current appeared in 70-90% of the neurons. 4. Five days after infection, expression of GABAC receptors in hippocampal neurons significantly decreased native GABAA receptor currents from 1200 ± 300 to 150 ± 70 pA (n = 10). The native glutamate-activated current was unchanged. 5. Hippocampal slices (P8) did not show a native GABAC-like current, although recombinant ρ1 receptors could be expressed in cultured hippocampal slices after adeno-ρ infection. 6. These data indicate that an adenovirus can be used to express recombinant GABAC receptors in hippocampal neurons. This finding could represent an important step towards the gene therapy of CNS receptor-related diseases.

Original languageEnglish (US)
Pages (from-to)145-153
Number of pages9
JournalJournal of Physiology
Volume535
Issue number1
DOIs
StatePublished - Aug 15 2001
Externally publishedYes

Fingerprint

Adenoviridae Infections
Human Adenoviruses
Neurons
Infection
HEK293 Cells
Adenoviridae
Cell Line
Antibodies
Patch-Clamp Techniques
GABA-A Receptors
Genetic Therapy
Transfection
Glutamic Acid
Western Blotting
GABA-C receptor
Staining and Labeling
Kidney

ASJC Scopus subject areas

  • Physiology

Cite this

Recombinant GABAC receptors expressed in rat hippocampal neurons after infection with an adenovirus containing the human ρ1 subunit. / Filippova, N.; Sedelnikova, A.; Tyler, William; Whitworth, T. L.; Fortinberry, H.; Weiss, D. S.

In: Journal of Physiology, Vol. 535, No. 1, 15.08.2001, p. 145-153.

Research output: Contribution to journalArticle

Filippova, N. ; Sedelnikova, A. ; Tyler, William ; Whitworth, T. L. ; Fortinberry, H. ; Weiss, D. S. / Recombinant GABAC receptors expressed in rat hippocampal neurons after infection with an adenovirus containing the human ρ1 subunit. In: Journal of Physiology. 2001 ; Vol. 535, No. 1. pp. 145-153.
@article{3d1461dff4d8461f92d1e88a6571be03,
title = "Recombinant GABAC receptors expressed in rat hippocampal neurons after infection with an adenovirus containing the human ρ1 subunit",
abstract = "1. A recombinant adenovirus was generated with the human ρ1 GABAC receptor subunit (adeno-ρ). Patch-clamp and antibody staining were employed to confirm functional expression of recombinant ρ1 receptors after infection of human embryonic kidney cells (HEK293 cell line), human embryonic retinal cells (911 cell line), dissociated rat hippocampal neurons and cultured rat hippocampal slices. 2. Standard whole-cell recording and Western blot analysis using ρ1 GABAC receptor antibodies revealed that recombinant ρ1 receptors were expressed in HEK293 and 911 cells after adeno-ρ infection and exhibited properties similar to those of ρ1 receptors after standard transfection. 3. Cultured rat hippocampal neurons (postnatal day (P)3-P5) did not show a native GABAC-like current. After adeno-ρ infection, however, a GABAC-like current appeared in 70-90{\%} of the neurons. 4. Five days after infection, expression of GABAC receptors in hippocampal neurons significantly decreased native GABAA receptor currents from 1200 ± 300 to 150 ± 70 pA (n = 10). The native glutamate-activated current was unchanged. 5. Hippocampal slices (P8) did not show a native GABAC-like current, although recombinant ρ1 receptors could be expressed in cultured hippocampal slices after adeno-ρ infection. 6. These data indicate that an adenovirus can be used to express recombinant GABAC receptors in hippocampal neurons. This finding could represent an important step towards the gene therapy of CNS receptor-related diseases.",
author = "N. Filippova and A. Sedelnikova and William Tyler and Whitworth, {T. L.} and H. Fortinberry and Weiss, {D. S.}",
year = "2001",
month = "8",
day = "15",
doi = "10.1111/j.1469-7793.2001.00145.x",
language = "English (US)",
volume = "535",
pages = "145--153",
journal = "Journal of Physiology",
issn = "0022-3751",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Recombinant GABAC receptors expressed in rat hippocampal neurons after infection with an adenovirus containing the human ρ1 subunit

AU - Filippova, N.

AU - Sedelnikova, A.

AU - Tyler, William

AU - Whitworth, T. L.

AU - Fortinberry, H.

AU - Weiss, D. S.

PY - 2001/8/15

Y1 - 2001/8/15

N2 - 1. A recombinant adenovirus was generated with the human ρ1 GABAC receptor subunit (adeno-ρ). Patch-clamp and antibody staining were employed to confirm functional expression of recombinant ρ1 receptors after infection of human embryonic kidney cells (HEK293 cell line), human embryonic retinal cells (911 cell line), dissociated rat hippocampal neurons and cultured rat hippocampal slices. 2. Standard whole-cell recording and Western blot analysis using ρ1 GABAC receptor antibodies revealed that recombinant ρ1 receptors were expressed in HEK293 and 911 cells after adeno-ρ infection and exhibited properties similar to those of ρ1 receptors after standard transfection. 3. Cultured rat hippocampal neurons (postnatal day (P)3-P5) did not show a native GABAC-like current. After adeno-ρ infection, however, a GABAC-like current appeared in 70-90% of the neurons. 4. Five days after infection, expression of GABAC receptors in hippocampal neurons significantly decreased native GABAA receptor currents from 1200 ± 300 to 150 ± 70 pA (n = 10). The native glutamate-activated current was unchanged. 5. Hippocampal slices (P8) did not show a native GABAC-like current, although recombinant ρ1 receptors could be expressed in cultured hippocampal slices after adeno-ρ infection. 6. These data indicate that an adenovirus can be used to express recombinant GABAC receptors in hippocampal neurons. This finding could represent an important step towards the gene therapy of CNS receptor-related diseases.

AB - 1. A recombinant adenovirus was generated with the human ρ1 GABAC receptor subunit (adeno-ρ). Patch-clamp and antibody staining were employed to confirm functional expression of recombinant ρ1 receptors after infection of human embryonic kidney cells (HEK293 cell line), human embryonic retinal cells (911 cell line), dissociated rat hippocampal neurons and cultured rat hippocampal slices. 2. Standard whole-cell recording and Western blot analysis using ρ1 GABAC receptor antibodies revealed that recombinant ρ1 receptors were expressed in HEK293 and 911 cells after adeno-ρ infection and exhibited properties similar to those of ρ1 receptors after standard transfection. 3. Cultured rat hippocampal neurons (postnatal day (P)3-P5) did not show a native GABAC-like current. After adeno-ρ infection, however, a GABAC-like current appeared in 70-90% of the neurons. 4. Five days after infection, expression of GABAC receptors in hippocampal neurons significantly decreased native GABAA receptor currents from 1200 ± 300 to 150 ± 70 pA (n = 10). The native glutamate-activated current was unchanged. 5. Hippocampal slices (P8) did not show a native GABAC-like current, although recombinant ρ1 receptors could be expressed in cultured hippocampal slices after adeno-ρ infection. 6. These data indicate that an adenovirus can be used to express recombinant GABAC receptors in hippocampal neurons. This finding could represent an important step towards the gene therapy of CNS receptor-related diseases.

UR - http://www.scopus.com/inward/record.url?scp=0035880813&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035880813&partnerID=8YFLogxK

U2 - 10.1111/j.1469-7793.2001.00145.x

DO - 10.1111/j.1469-7793.2001.00145.x

M3 - Article

C2 - 11507165

AN - SCOPUS:0035880813

VL - 535

SP - 145

EP - 153

JO - Journal of Physiology

JF - Journal of Physiology

SN - 0022-3751

IS - 1

ER -