| First Author | Deshpande A | Year | 2020 |
| Journal | Biomolecules | Volume | 10 |
| Issue | 3 | PubMed ID | 32204458 |
| Mgi Jnum | J:304046 | Mgi Id | MGI:6693968 |
| Doi | 10.3390/biom10030470 | Citation | Deshpande A, et al. (2020) Why Does Knocking Out NACHO, But Not RIC3, Completely Block Expression of alpha7 Nicotinic Receptors in Mouse Brain?. Biomolecules 10(3):470 |
| abstractText | Alpha7 nicotinic acetylcholine receptors (alpha7nAChRs) are interesting not only because of their physiological effects, but because this receptor requires chaperones to traffic to cell surfaces (measured by alpha-bungarotoxin [alphaBGT] binding). While knockout (KO) animals and antibodies that react across species exist for tmem35a encoding the protein chaperone NACHO, commercially available antibodies against the chaperone RIC3 that allow Western blots across species have not been generally available. Further, no effects of deleting RIC3 function (ric3 KO) on alpha7nAChR expression are reported. Finally, antibodies against alpha7nAChRs have shown various deficiencies. We find mouse macrophages bind alphaBGT but lack NACHO. We also report on a new alpha7nAChR antibody and testing commercially available anti-RIC3 antibodies that react across species allowing Western blot analysis of in vitro cultures. These antibodies also react to specific RIC3 splice variants and single-nucleotide polymorphisms. Preliminary autoradiographic analysis reveals that ric3 KOs show subtle alphaBGT binding changes across different mouse brain regions, while tmem35a KOs show a complete loss of alphaBGT binding. These findings are inconsistent with effects observed in vitro, as RIC3 promotes alphaBGT binding to alpha7nAChRs expressed in HEK cells, even in the absence of NACHO. Collectively, additional regulatory factors are likely involved in the in vivo expression of alpha7nAChRs. |
