Introduction: Binding assays are essential in the screening of substances for new drugs, allowing the evaluation of the interaction of compounds with relevant molecular targets. These assays are applicable to the rational design of drugs and the investigation of interactions between neurotransmitters and receptors. Given the characteristics of Autism Spectrum Disorder (ASD) and the scarcity of therapeutic options, the development of new approaches, such as the investigation of bioinspired peptides like NeuroVAL, becomes urgent. The objective of this study was to standardize binding assays in dopaminergic receptors to evaluate NeuroVAL in a model of ASD induced by valproic acid. Methodology: The assays performed included competition, saturation, and uptake, utilizing tritium linked to dopamine (3H-dopamine). Parameters were determined for the extraction of synaptosomes and neural membranes from female Swiss mice, following approval by the Animal Ethics Committee (CEUA) of the University of Brasília, under protocol numbers 23106.147095/2018-89 and 23106.039800/2024-13. Parameters for the assays were also established, such as time, temperature, and buffer, ensuring controlled conditions. Results: The results demonstrated the importance of standardizing assays to quantify receptors in brain material from animals with ASD induced by VPA. The assays allowed for the evaluation of NeuroVAL’s action, although the identified non-specific binding requires experimental refinement to improve data reliability. Conclusions: This study standardized parameters for binding assays in dopaminergic receptors, aiming for application in a model of ASD induced by VPA. This standardization is crucial for enabling the precise quantification of neural receptors and contributing to the development of new treatments that improve the quality of life for patients.
Author Contributions
J.W.F.: Visualization, methodology, investigation, writing; G.A.A.C.: Visualization, methodology, investigation, writing, formal analysis; L.G.V.: Visualization, methodology; M.R.M.: Writing—review & editing, supervision, funding acquisition, conceptualization. All authors have read and agreed to the published version of the manuscript.
Funding
This research was funded by the Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) under grant number 00193.0000724/2021-30.
Institutional Review Board Statement
The animal study protocol was approved by the Animal Ethics Committee (CEUA) of the University of Brasília, under protocol numbers 23106.147095/2018-89 and 23106.039800/2024-13.
Informed Consent Statement
Not applicable.
Data Availability Statement
The datasets presented in this article are not readily available because the data are part of an ongoing study. Requests to access the datasets should be directed to jessicaweschenfelderjwf@gmail.com.
Conflicts of Interest
The authors declare no conflict of interest.
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