Knockout or Knock-in? A Truncated D2 Receptor Protein Is Expressed in the Brain of Functional D2 Receptor Knockout Mice
| Autor: | Roberto Munita, Angélica P. Escobar, Natalia Sánchez, Mauricio Herrera-Rojas, María Estela Andrés, Rodrigo Meza, Montserrat Olivares-Costa, Marcela González, Jessica Albornoz, Gianluca Merello |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
truncated protein Endoplasmic reticulum dopamine 2 receptor functional knockout Neurosciences. Biological psychiatry. Neuropsychiatry Biology Golgi apparatus Cell biology 03 medical and health sciences Transmembrane domain symbols.namesake 030104 developmental biology 0302 clinical medicine Quinpirole Dopamine receptor D2 Complementary DNA Gene knockin Knockout mouse medicine symbols 030217 neurology & neurosurgery medicine.drug RC321-571 |
| Zdroj: | NeuroSci, Vol 2, Iss 14, Pp 193-206 (2021) NeuroSci Volume 2 Issue 2 Pages 14-206 |
| ISSN: | 2673-4087 |
| Popis: | Null mice for the dopamine D2 receptor (D2R) have been instrumental in understanding the function of this protein. For our research, we obtained the functional D2R knockout mouse strain described initially in 1997. Surprisingly, our biochemical characterization showed that this mouse strain is not a true knockout. We determined by sequence analysis of the rapid 3′ amplification of cDNA ends that functional D2R knockout mice express transcripts that lack only the eighth exon. Furthermore, immunofluorescence assays showed a D2R-like protein in the brain of functional D2R knockout mice. We verified by immunofluorescence that the recombinant truncated D2R is expressed in HEK293T cells, showing intracellular localization, colocalizing in the Golgi apparatus and the endoplasmic reticulum, but with less presence in the Golgi apparatus compared to the native D2R. As previously reported, functional D2R knockout mice are hypoactive and insensitive to the D2R agonist quinpirole. Concordantly, microdialysis studies confirmed that functional D2R knockout mice have lower extracellular dopamine levels in the striatum than the native mice. In conclusion, functional D2R knockout mice express transcripts that lead to a truncated D2R protein lacking from the sixth transmembrane domain to the C-terminus. We share these findings to avoid future confusion and the community considers this mouse strain in D2R traffic and protein–protein interaction studies. |
| Databáze: | OpenAIRE |
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