Identification of Carassius auratus gibelio liver cell proteins interacting with the GABAA receptor γ2 subunit using a yeast two-hybrid system
The γ-aminobutyric acid type A (GABAA) receptor is an important pentameric inhibitory neurotransmitter receptor, and the γ2 subunit of this receptor plays a key role in potentiation of the GABAA response. We previously detected that the expression of GABAA receptor in the livers of Carassius auratus gibelio significantly increased after medication (avermectin and difloxacin treatment). In order to better understand the mechanism of action of the GABAA receptor γ2 subunit in the livers of C. auratus gibelio, we constructed a C. auratus gibelio liver cDNA library (the titer value of 1.2 × 106 cfu/mL) and identified the proteins that interact with the GABAA receptor γ2 subunit by using a yeast two-hybrid assay. The yeast two-hybrid screening yielded seven positive clones, namely, prelid3b, cdc42, sgk1, spg21, proteasome, chia.5, and AP-3 complex subunit beta-1, all of which have been annotated by the NCBI database. The functions of these proteins are complex; therefore, additional studies are required to determine the specific interactions of these proteins with the GABAA receptor γ2 subunit in the liver of C. auratus gibelio. Although the interactions identified by the yeast two-hybrid system should be considered as preliminary results, the findings of this study may provide further direction and a foundation for future research focusing on the mechanisms of the GABAA receptor γ2 subunit in C. auratus gibelio livers.
KeywordsCarassius auratus gibelio GABAA receptor γ2 subunit Liver cDNA library Yeast two-hybrid Protein–protein interaction
This study was supported by the Special Fund for Agro-scientific Research in the Public Interest (Grant 201203085), the 863 Program (Grant 2011AA10A216), the National Natural Resources Platform, and the Shanghai University Knowledge Service Platform.
Compliance with ethical standards
Experimental procedures were carried out in accordance with the Shanghai Ocean University Animal Health Guidelines for Animal Care and Experimentation and supervised by the Institutional Animal Care and Use Committee at Shanghai Ocean University.
Conflict of interest
The authors declare that they have no conflicts of interest.
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