Caspase-3 and GFAP as early markers for apoptosis and astrogliosis in shRNA-induced hippocampal cytotoxicity

Anne Günther; Vince Luczak; Ted Abel; Arnd Baumann

doi:10.1242/jeb.154583

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Caspase-3 and GFAP as early markers for apoptosis and astrogliosis in shRNA-induced hippocampal cytotoxicity

Journal article

Open access

Peer reviewed

Caspase-3 and GFAP as early markers for apoptosis and astrogliosis in shRNA-induced hippocampal cytotoxicity

Anne Günther, Vince Luczak, Ted Abel and Arnd Baumann

Journal of experimental biology, Vol.220(Pt 8), pp.1400-1404

04/15/2017

DOI: 10.1242/jeb.154583

PMCID: PMC5413069

PMID: 28167801

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Abstract

Genetic manipulation of cells and tissue by RNA interference has significantly contributed to the functional characterization of individual proteins and their role in physiological processes. Despite its versatility, RNA interference can have detrimental side effects, including reduced cell viability. We applied recombinant adeno-associated viruses by stereotaxic injection into the murine hippocampus to express different short hairpin RNA (shRNA) constructs along with eGFP. Tissue responses were assessed immunohistochemically for up to 8 weeks post-infection. Strong hippocampal degeneration and tissue atrophy was observed, most likely induced by high shRNA expression. The effect was entirely absent in mice injected with vectors driving only expression of eGFP. Active caspase-3 (Casp-3) and glial fibrillary acidic protein (GFAP) were identified as molecular markers and early indicators of adverse tissue responses. Our findings also demonstrate that detrimental effects of high shRNA expression in hippocampal tissue can be monitored even before the onset of tissue degeneration.

Dependovirus - genetics

Neurons - pathology

RNA, Small Interfering - genetics

Gene Expression

Glial Fibrillary Acidic Protein - analysis

Humans

Mice, Inbred C57BL

Cells, Cultured

RNA, Small Interfering - adverse effects

Male

Green Fluorescent Proteins - genetics

Hippocampus - pathology

Hippocampus - metabolism

Animals

RNA Interference

Caspase 3 - analysis

Mice

Neurons - metabolism

RNA, Small Interfering - administration & dosage

Apoptosis

Details

Title: Subtitle: Caspase-3 and GFAP as early markers for apoptosis and astrogliosis in shRNA-induced hippocampal cytotoxicity
Creators: Anne Günther - Institute of Complex Systems, Cellular Biophysics (ICS-4), Forschungszentrum Jülich, Jülich 52425, Germany
Vince Luczak - Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA
Ted Abel - Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA
Arnd Baumann - Institute of Complex Systems, Cellular Biophysics (ICS-4), Forschungszentrum Jülich, Jülich 52425, Germany a.baumann@fz-juelich.de
Resource Type: Journal article
Publication Details: Journal of experimental biology, Vol.220(Pt 8), pp.1400-1404
Publisher: England
DOI: 10.1242/jeb.154583
PMID: 28167801
PMCID: PMC5413069
ISSN: 0022-0949
eISSN: 1477-9145
Grant note: R01 MH087463 / NIMH NIH HHS R01 DA036984 / NIDA NIH HHS R01 MH099544 / NIMH NIH HHS R01 MH101491 / NIMH NIH HHS
Language: English
Date published: 04/15/2017
Academic Unit: Molecular Physiology and Biophysics; Psychiatry; Psychological and Brain Sciences; Iowa Neuroscience Institute; Neuroscience and Pharmacology; Biochemistry and Molecular Biology
Record Identifier: 9984065823702771

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