Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters
OCT1 and OCT2 are polyspecific membrane transporters that are involved in hepatic and renal drug clearance in humans and mice. In this study, we cloned dog OCT1 and OCT2 and compared their function to the human and mouse orthologs. We used liver and kidney RNA to clone dog OCT1 and OCT2. The cloned...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/9/5100 |
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| author | Marleen Julia Meyer Simon Falk Sarah Römer Clarissa Prinzinger Sabine Tacke Joachim Geyer Stefan Simm Mladen Vassilev Tzvetkov |
| author_facet | Marleen Julia Meyer Simon Falk Sarah Römer Clarissa Prinzinger Sabine Tacke Joachim Geyer Stefan Simm Mladen Vassilev Tzvetkov |
| author_sort | Marleen Julia Meyer |
| collection | DOAJ |
| description | OCT1 and OCT2 are polyspecific membrane transporters that are involved in hepatic and renal drug clearance in humans and mice. In this study, we cloned dog OCT1 and OCT2 and compared their function to the human and mouse orthologs. We used liver and kidney RNA to clone dog OCT1 and OCT2. The cloned and the publicly available RNA-Seq sequences differed from the annotated exon-intron structure of OCT1 in the dog genome CanFam3.1. An additional exon between exons 2 and 3 was identified and confirmed by sequencing in six additional dog breeds. Next, dog OCT1 and OCT2 were stably overexpressed in HEK293 cells and the transport kinetics of five drugs were analyzed. We observed strong differences in the transport kinetics between dog and human orthologs. Dog OCT1 transported fenoterol with 12.9-fold higher capacity but 14.3-fold lower affinity (higher K<sub>M</sub>) than human OCT1. Human OCT1 transported ipratropium with 5.2-fold higher capacity but 8.4-fold lower affinity than dog OCT1. Compared to human OCT2, dog OCT2 showed 10-fold lower transport of fenoterol and butylscopolamine. In conclusion, the functional characterization of dog OCT1 and OCT2 reported here may have implications when using dogs as pre-clinical models as well as for drug therapy in dogs. |
| first_indexed | 2024-03-10T04:04:05Z |
| format | Article |
| id | doaj.art-300804902216439392bb945ae7d65708 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:04:05Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-300804902216439392bb945ae7d657082023-11-23T08:27:07ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-01239510010.3390/ijms23095100Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation TransportersMarleen Julia Meyer0Simon Falk1Sarah Römer2Clarissa Prinzinger3Sabine Tacke4Joachim Geyer5Stefan Simm6Mladen Vassilev Tzvetkov7Department of General Pharmacology, Institute of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, 17487 Greifswald, GermanyDepartment of General Pharmacology, Institute of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, 17487 Greifswald, GermanyDepartment of General Pharmacology, Institute of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, 17487 Greifswald, GermanyFaculty of Veterinary Medicine, Institute of Pharmacology and Toxicology, Justus Liebig University Giessen, 35390 Giessen, GermanyClinic for Small Animals, Faculty of Veterinary Medicine, Justus Liebig University Giessen, 35390 Giessen, GermanyFaculty of Veterinary Medicine, Institute of Pharmacology and Toxicology, Justus Liebig University Giessen, 35390 Giessen, GermanyInstitute of Bioinformatics, University Medicine Greifswald, 17487 Greifswald, GermanyDepartment of General Pharmacology, Institute of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, 17487 Greifswald, GermanyOCT1 and OCT2 are polyspecific membrane transporters that are involved in hepatic and renal drug clearance in humans and mice. In this study, we cloned dog OCT1 and OCT2 and compared their function to the human and mouse orthologs. We used liver and kidney RNA to clone dog OCT1 and OCT2. The cloned and the publicly available RNA-Seq sequences differed from the annotated exon-intron structure of OCT1 in the dog genome CanFam3.1. An additional exon between exons 2 and 3 was identified and confirmed by sequencing in six additional dog breeds. Next, dog OCT1 and OCT2 were stably overexpressed in HEK293 cells and the transport kinetics of five drugs were analyzed. We observed strong differences in the transport kinetics between dog and human orthologs. Dog OCT1 transported fenoterol with 12.9-fold higher capacity but 14.3-fold lower affinity (higher K<sub>M</sub>) than human OCT1. Human OCT1 transported ipratropium with 5.2-fold higher capacity but 8.4-fold lower affinity than dog OCT1. Compared to human OCT2, dog OCT2 showed 10-fold lower transport of fenoterol and butylscopolamine. In conclusion, the functional characterization of dog OCT1 and OCT2 reported here may have implications when using dogs as pre-clinical models as well as for drug therapy in dogs.https://www.mdpi.com/1422-0067/23/9/5100organic cation transporterSLC22A1SLC22A2species differencesortholog comparisongene structure |
| spellingShingle | Marleen Julia Meyer Simon Falk Sarah Römer Clarissa Prinzinger Sabine Tacke Joachim Geyer Stefan Simm Mladen Vassilev Tzvetkov Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters International Journal of Molecular Sciences organic cation transporter SLC22A1 SLC22A2 species differences ortholog comparison gene structure |
| title | Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters |
| title_full | Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters |
| title_fullStr | Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters |
| title_full_unstemmed | Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters |
| title_short | Cloning and Functional Characterization of Dog OCT1 and OCT2: Another Step in Exploring Species Differences in Organic Cation Transporters |
| title_sort | cloning and functional characterization of dog oct1 and oct2 another step in exploring species differences in organic cation transporters |
| topic | organic cation transporter SLC22A1 SLC22A2 species differences ortholog comparison gene structure |
| url | https://www.mdpi.com/1422-0067/23/9/5100 |
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