Between Dec 19, 2024 and Jan 2, 2025, datasets can be submitted to DRUM but will not be processed until after the break. Staff will not be available to answer email during this period, and will not be able to provide DOIs until after Jan 2. If you are in need of a DOI during this period, consider Dryad or OpenICPSR. Submission responses to the UDC may also be delayed during this time.
 

Verification of AMPK Knockdown in Sprague Dawley Rats using a Vivo-Morpholino Antisense Oligonucleotide

Loading...
Thumbnail Image

Persistent link to this item

Statistics
View Statistics

Journal Title

Journal ISSN

Volume Title

Title

Verification of AMPK Knockdown in Sprague Dawley Rats using a Vivo-Morpholino Antisense Oligonucleotide

Published Date

2024-01-09

Publisher

Type

Presentation

Abstract

Metformin, a drug typically used to treat type II diabetes, can decrease cue-induced cocaine seeking in rats. The proposed mechanism of action for this effect is the activation of adenosine monophosphate-activated protein kinase (AMPK). To confirm this mechanism for the effect of metformin we are validating a technique for knocking down AMPK in the nucleus accumbens core (NAcC) in vivo. This study tests whether microinjections of a vivo-morpholino antisense oligonucleotide (VM-ASO) in the NAcC is an effective method of knocking down AMPK in Sprague Dawley rats. VM-ASO binds target mRNA and prevents translation. This experiment used 8 Sprague Dawley rats (6 male, 2 female). Initially, rats underwent stereotaxic intracranial surgery for the insertion of a cannula into both hemispheres of the NAcC (from bregma: AP +1.5; ML: +/- 1.8; DV: - 5.5mm). Then, 140 pmol of AMPK VM-ASO or scrambled control nucleotide was microinjected in each hemisphere at a rate of 0.2 µl/minute 2mm below the base of the cannula. After a one-minute diffusion period, the microinjectors were removed. This process was repeated for four days. Eight days after the last microinjection, tissue samples were collected and AMPK levels were analyzed via western blot. It was found that microinjections of AMPK VM-ASO did not significantly decrease AMPK levels in the NAcC.

Description

Related to

Replaces

License

Series/Report Number

Funding information

This research was supported by the Undergraduate Research Opportunities Program (UROP).

Isbn identifier

Doi identifier

Previously Published Citation

Other identifiers

Suggested citation

Hunsader, Peter; Hernandez, Edith; Moore, Maddi; Slosky, Lauren; Spencer, Sade. (2024). Verification of AMPK Knockdown in Sprague Dawley Rats using a Vivo-Morpholino Antisense Oligonucleotide. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/259935.

Content distributed via the University Digital Conservancy may be subject to additional license and use restrictions applied by the depositor. By using these files, users agree to the Terms of Use. Materials in the UDC may contain content that is disturbing and/or harmful. For more information, please see our statement on harmful content in digital repositories.