Cargando…
Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell
We demonstrate here that highly sensitive in vitro bioassays for FSH, TSH, and PTH can be set up in mouse Leydig Tumor Cells (mLTC), in addition to the normal LH/CG bioassay, after they were transfected with expression vectors encoding the corresponding Gs Protein-Coupled Receptors (GsPCR), such as...
Autores principales: | , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10419024/ https://www.ncbi.nlm.nih.gov/pubmed/37569429 http://dx.doi.org/10.3390/ijms241512047 |
_version_ | 1785088410788036608 |
---|---|
author | Klett, Danièle Pellissier, Lucie Lomet, Didier Derouin-Tochon, Flavie Robert, Vincent Nguyen, Thi Mong Diep Duittoz, Anne Reiter, Eric Locatelli, Yann Dupont, Joëlle Dardente, Hugues Jean-Alphonse, Frédéric Combarnous, Yves |
author_facet | Klett, Danièle Pellissier, Lucie Lomet, Didier Derouin-Tochon, Flavie Robert, Vincent Nguyen, Thi Mong Diep Duittoz, Anne Reiter, Eric Locatelli, Yann Dupont, Joëlle Dardente, Hugues Jean-Alphonse, Frédéric Combarnous, Yves |
author_sort | Klett, Danièle |
collection | PubMed |
description | We demonstrate here that highly sensitive in vitro bioassays for FSH, TSH, and PTH can be set up in mouse Leydig Tumor Cells (mLTC), in addition to the normal LH/CG bioassay, after they were transfected with expression vectors encoding the corresponding Gs Protein-Coupled Receptors (GsPCR), such as FSHR, TSHR, or PTHR. Although the β2 adrenergic receptor is also a GsPCR, its expression in mLTC led to a significant but very low cAMP response compared to those observed with FSH, TSH, or PTH. Similarly, after transfection of the GiPCR MT1 melatonin receptor, we did not observe any inhibitory effect by melatonin of the LH or hCG stimulation. Interestingly, after transfection of mLTC with the human kisspeptin receptor (hKpR), which is a GqPCR, we observed a dose-dependent synergy of 10(−12)–10(−7) M kisspeptin variants with a fixed concentration of 0.3 nM LH or hCG. Without any exogenous receptor transfection, a 2 h preincubation with OT or AVP led to a dose-dependent cAMP response to a fixed dose of LH or hCG. Therefore, highly sensitive in vitro bioassays for various hormones and other GPCR ligands can be set up in mLTC to measure circulating concentrations in only 3–10 µL of blood or other body fluids. Nevertheless, the development of an LHRKO mLTC cell line will be mandatory to obtain strict specificity for these bioassays to eliminate potential cross-reaction with LH or CG. |
format | Online Article Text |
id | pubmed-10419024 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-104190242023-08-12 Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell Klett, Danièle Pellissier, Lucie Lomet, Didier Derouin-Tochon, Flavie Robert, Vincent Nguyen, Thi Mong Diep Duittoz, Anne Reiter, Eric Locatelli, Yann Dupont, Joëlle Dardente, Hugues Jean-Alphonse, Frédéric Combarnous, Yves Int J Mol Sci Article We demonstrate here that highly sensitive in vitro bioassays for FSH, TSH, and PTH can be set up in mouse Leydig Tumor Cells (mLTC), in addition to the normal LH/CG bioassay, after they were transfected with expression vectors encoding the corresponding Gs Protein-Coupled Receptors (GsPCR), such as FSHR, TSHR, or PTHR. Although the β2 adrenergic receptor is also a GsPCR, its expression in mLTC led to a significant but very low cAMP response compared to those observed with FSH, TSH, or PTH. Similarly, after transfection of the GiPCR MT1 melatonin receptor, we did not observe any inhibitory effect by melatonin of the LH or hCG stimulation. Interestingly, after transfection of mLTC with the human kisspeptin receptor (hKpR), which is a GqPCR, we observed a dose-dependent synergy of 10(−12)–10(−7) M kisspeptin variants with a fixed concentration of 0.3 nM LH or hCG. Without any exogenous receptor transfection, a 2 h preincubation with OT or AVP led to a dose-dependent cAMP response to a fixed dose of LH or hCG. Therefore, highly sensitive in vitro bioassays for various hormones and other GPCR ligands can be set up in mLTC to measure circulating concentrations in only 3–10 µL of blood or other body fluids. Nevertheless, the development of an LHRKO mLTC cell line will be mandatory to obtain strict specificity for these bioassays to eliminate potential cross-reaction with LH or CG. MDPI 2023-07-27 /pmc/articles/PMC10419024/ /pubmed/37569429 http://dx.doi.org/10.3390/ijms241512047 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Klett, Danièle Pellissier, Lucie Lomet, Didier Derouin-Tochon, Flavie Robert, Vincent Nguyen, Thi Mong Diep Duittoz, Anne Reiter, Eric Locatelli, Yann Dupont, Joëlle Dardente, Hugues Jean-Alphonse, Frédéric Combarnous, Yves Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell |
title | Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell |
title_full | Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell |
title_fullStr | Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell |
title_full_unstemmed | Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell |
title_short | Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell |
title_sort | highly-sensitive in vitro bioassays for fsh, tsh, pth, kp, and ot in addition to lh in mouse leydig tumor cell |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10419024/ https://www.ncbi.nlm.nih.gov/pubmed/37569429 http://dx.doi.org/10.3390/ijms241512047 |
work_keys_str_mv | AT klettdaniele highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT pellissierlucie highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT lometdidier highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT derouintochonflavie highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT robertvincent highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT nguyenthimongdiep highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT duittozanne highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT reitereric highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT locatelliyann highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT dupontjoelle highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT dardentehugues highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT jeanalphonsefrederic highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell AT combarnousyves highlysensitiveinvitrobioassaysforfshtshpthkpandotinadditiontolhinmouseleydigtumorcell |