PhD Scientific Days 2024

Poster Session B - Pharmaceutical Sciences and Health Technologies 1.

Optimization of a synaptosome model to study the effects of inhibition of angiotensin receptor 1 and  2 on synaptic glutamate release

Előadó neve

Mr. Lincmajer, Zoltan

Neptun code

AJURB4

Előadó munkahelye

Department of pharmacodynamics

Előadó telefonszáma

+3606708594

Előadó e-mail címe

lincmajer.zoltan@stud.semmelweis.hu

Az előadás címe

Optimization of a synaptosome model to study the effects of inhibition of angiotensin receptor 1 and  2 on synaptic glutamate release

Szerző(k) neve és munkahelye

Zoltan Lincmajer¹

1: Department of Pharmacodynamics

Bemutatás módja

Poszter

Szekció

Poster Session B - Pharmaceutical Sciences and Health Technologies 1.

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction:
Central nervous system effects of angiotensin receptor blockers (ARBs) gained a massive interest recently. Previous findings from our research group have demonstrated the anti-allodynic effect of ARBs in neuropathic animal model. Furthermore, telmisartan was found capable of enhancing the antinociceptive effect of morphine and reducing opioid tolerance.

Aims:
In this study, we intended to examine the impact of ARBs on synaptic glutamate transmission using a synaptosome model. Our aim was to transfer the previously applied synaptosome model into a suspension format and its optimization for a more sensitive and robust assessment of neurotransmitter release. We aimed at investigating the effect of ARBs on synaptic glutamate release using the optimized method.

Methods:
Freshly prepared synaptosomes and cryopreserved ones were examined. Their viability was assessed using resazurin reduction and LDH release assays. The amount of released glutamate was quantified using a glutamate oxidase enzyme-coupled reaction with fluorescence detection method.

Results:
Our results indicated that synaptosomes maintained their viability following resuspendation. Compared to the previously applied method significantly lower amount of synaptosome can be used, as approximately 3 µM of glutamate was released from 2 mg of synaptosomes. Suspension method is characterized by a more uniform distribution of synaptosomes resulting more reproducible and robust assay. Based on the results of storage stability experiments of freshly prepared suspension, viability decline can be avoided using snap freezing in HEPES-sucrose buffer containing 10% FBS without addition of DMSO.
Using the optimized method, we examined the effect of ARBs on glutamate release, showing a concentration-dependent reduction in 4-aminopyridine-induced glutamate release.
Interestingly angiotensin II itself could not increase the glutamate release suggesting the involvement of multiple receptors in its effect.

Conclusions:
The optimized model provides a more physiological method for studying the effects of various compounds on neurotransmitter release. This approach significantly reduces the required tissue quantity, thereby minimizing the number of animals to be sacrificed. Reduced glutamate release in response to ARBs may contribute to their previously observed antinociceptive effect.

University

Semmelweis University

Supervisor

Dr. Tabi Tamas

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

8081

Start

15:30

End

15:33