Journal article
Stereotactic Delivery of Helper-dependent Adenoviral Viral Vectors at Distinct Developmental Time Points to Perform Age-dependent Molecular Manipulations of the Mouse Calyx of Held
Bio-protocol, Vol.13(16), e4793
08/20/2023
DOI: 10.21769/BioProtoc.4793
PMCID: PMC10450731
PMID: 37638292
Abstract
Synapses are specialized structures that enable neuronal communication, which is essential for brain function and development. Alterations in synaptic proteins have been linked to various neurological and neuropsychiatric disorders. Therefore, manipulating synaptic proteins in vivo can provide insight into the molecular mechanisms underlying these disorders and aid in developing new therapeutic strategies. Previous methods such as constitutive knock-out animals are limited by developmental compensation and off-target effects. The current approach outlines procedures for age-dependent molecular manipulations in mice using helper-dependent adenovirus viral vectors (HdAd) at distinct developmental time points. Using stereotactic injection of HdAds in both newborn and juvenile mice, we demonstrate the versatility of this method to express Cre recombinase in globular bushy cells of juvenile
Rac1
fl/fl
mice to ablate presynaptic Rac1 and study its role in synaptic transmission. Separately, we overexpress Ca
V
2 α1 subunits at two distinct developmental time points to elucidate the mechanisms that determine presynaptic Ca
V
2 channel abundance and preference. This method presents a reliable, cost-effective, and minimally invasive approach for controlling gene expression in specific regions of the mouse brain and will be a powerful tool to decipher brain function in health and disease.
Key features
Virus-mediated genetic perturbation in neonatal and young adult mice.
Stereotaxic injection allows targeting of brain structures at different developmental stages to study the impact of genetic perturbation throughout the development.
Details
- Title: Subtitle
- Stereotactic Delivery of Helper-dependent Adenoviral Viral Vectors at Distinct Developmental Time Points to Perform Age-dependent Molecular Manipulations of the Mouse Calyx of Held
- Creators
- Christian Keine - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA, USA Department of Human Medicine, University of Oldenburg, Oldenburg, Germany Research Center Neurosensory Science, Oldenburg, GermanyMohammed Al-Yaari - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA, USATamara Radulovic - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA, USA Department of Human Medicine, University of Oldenburg, Oldenburg, Germany Research Center Neurosensory Science, Oldenburg, GermanySamuel M. Young - Department of Anatomy and Cell Biology, University of Iowa, Iowa City, IA, USA Department of Otolaryngology, Iowa Neuroscience Institute, University of Iowa, Iowa City, IA, USA
- Resource Type
- Journal article
- Publication Details
- Bio-protocol, Vol.13(16), e4793
- Publisher
- Bio-Protocol
- DOI
- 10.21769/BioProtoc.4793
- PMID
- 37638292
- PMCID
- PMC10450731
- eISSN
- 2331-8325
- Language
- English
- Date published
- 08/20/2023
- Academic Unit
- Anatomy and Cell Biology; Iowa Neuroscience Institute; Otolaryngology
- Record Identifier
- 9984457960202771
Metrics
7 Record Views