Keine, Christian and Al-Yaari, Mohammed and Radulovic, Tamara and Young Jr., Samuel M. (2023) 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, 13 (16). pp. 1-11. ISSN 2331-8325

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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 Rac1fl/fl mice to ablate presynaptic Rac1 and study its role in synaptic transmission. Separately, we overexpress CaV2 α1 subunits at two distinct developmental time points to elucidate the mechanisms that determine presynaptic CaV2 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.

Item Type: Article
Uncontrolled Keywords: Calyx of Held, Synaptic transmission, Rac1, Presynaptic mechanisms, Helper-dependent adenovirus, Stereotactic injection
Subjects: Science and mathematics > Life sciences, biology
Divisions: Faculty of Medicine and Health Sciences > Department of Human Medicine
Date Deposited: 27 Sep 2023 10:59
Last Modified: 27 Sep 2023 10:59
URN: urn:nbn:de:gbv:715-oops-59285
DOI: 10.21769/BioProtoc.4793

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