Human DENND1A.V2 Drives Cyp17a1 Expression and Androgen Production in Mouse Ovaries and Adrenals
Abstract
:1. Introduction
2. Results
2.1. hDENND1A.V2 Induces a PCOS Phenotype in Mouse Leydig MA-10 Cells—Evidence to Support a hDENND1A.V2 Mouse Model for PCOS
2.2. Generation of hDENND1A.V2 Transgenic Mice
3. Discussion
4. Materials and Methods
4.1. Adenoviral Expression of hDENND1A in Mouse MA-10 Leydig Cells
4.2. Animals
4.3. Generation of CMV-hDENND1A.V2 Transgenic Mice
4.4. Generation of Lhcgr-hDENND1A.V2 Transgenic Mice
4.5. Generation of TetOn-hDENND1A.V2 Transgenic Mice
4.6. RT-PCR
4.7. Quantitative Real-Time qRT-PCR Analyses of DENND1A.V2 and Cyp17a1
4.8. Western Blot and Immunodetection
4.9. Cell Transfection
4.10. Immunofluorescence Detection of hDENND1A.V2
4.11. Immunohistochemical Detection of hDENND1A.V2
4.12. Theca-Interstitial Cell (TIC) and Granulosa Cell (GC) Isolation
4.13. Quantitation of Progesterone (P4), 17α-hydroxyprogesterone (17OHP4), and Androstenedione (adione) by ELISA
4.14. Statistical Methods
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
PCOS | polycystic ovary syndrome |
GWAS | Genome Wide Association Study |
DENND1A | DENN (Differentially Expressed in Neoplastic vs. Normal cells) Domain Containing 1A |
hDENND1A.V2 | hDENND1A variant 2 (a truncated splice variant of the DENND1A gene) |
Lhcgr | Luteinizing Hormone/Choriogonadotropin Receptor |
Cyp17 | mouse cytochrome P450 17-hydroxylase |
P4 | progesterone |
17OHP4 | 17-hydroxyprogesterone |
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Teves, M.E.; Modi, B.P.; Kulkarni, R.; Han, A.X.; Marks, J.S.; Subler, M.A.; Windle, J.; Newall, J.M.; McAllister, J.M.; Strauss, J.F., III. Human DENND1A.V2 Drives Cyp17a1 Expression and Androgen Production in Mouse Ovaries and Adrenals. Int. J. Mol. Sci. 2020, 21, 2545. https://doi.org/10.3390/ijms21072545
Teves ME, Modi BP, Kulkarni R, Han AX, Marks JS, Subler MA, Windle J, Newall JM, McAllister JM, Strauss JF III. Human DENND1A.V2 Drives Cyp17a1 Expression and Androgen Production in Mouse Ovaries and Adrenals. International Journal of Molecular Sciences. 2020; 21(7):2545. https://doi.org/10.3390/ijms21072545
Chicago/Turabian StyleTeves, Maria E., Bhavi P. Modi, Rewa Kulkarni, Angela X. Han, Jamaia S. Marks, Mark A. Subler, Jolene Windle, Jordan M. Newall, Jan M. McAllister, and Jerome F. Strauss, III. 2020. "Human DENND1A.V2 Drives Cyp17a1 Expression and Androgen Production in Mouse Ovaries and Adrenals" International Journal of Molecular Sciences 21, no. 7: 2545. https://doi.org/10.3390/ijms21072545