Expression of horse and donkey LH in COS-7 cells: evidence for low FSH activity in donkey LH compared with horse LH.
Abstract: Horse (Equus caballus) luteinizing hormone (eLH) and chorionic gonadotrophin (eCG), which have the same amino acid sequence, are unusual in that, although they express only LH activity in equids, they express dual LH and FSH activities in all other species tested. Donkey (Equus asinus) LH (dkLH) and CG (dkCG), which also share an identical peptide backbone, have been less well characterized and conflicting results concerning their FSH activity in heterologous species have appeared in the literature. In order to assess and compare the intrinsic LH and FSH activities of the horse and donkey LHs in heterologous species, recombinant eLH (r.eLH/CG) and recombinant dkLH (r.dkLH/CG) were expressed, for the first time, in COS-7 cells. Their LH activities were assessed in a rat Leydig cell bioassay, and their FSH activities were estimated in a bioassay using Y1 cells stably expressing the human FSH receptor. Human CG (hCG) was expressed (r.hCG) and analysed in the same system. The results showed that, whereas r.dkLH/CG was about twice as active as r.eLH/CG in the LH bioassay, it was five times less active than r.eLH/CG in the FSH bioassay; r.hCG was about three times less active than r.eLH/CG in the LH bioassay but was completely inactive in the FSH bioassay. These results confirm that dkLH/CG possesses significant FSH activity in heterologous species that is not attributable to contamination with FSH.
Publication Date: 1997-03-01 PubMed ID: 9071957DOI: 10.1677/joe.0.1520371Google Scholar: Lookup
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- Comparative Study
- Journal Article
Summary
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The research article discusses a study conducted to compare the luteinizing hormone (LH) and follicle-stimulating hormone (FSH) activities of horses and donkeys. The findings highlight a significantly lower FSH activity in donkeys compared to horses, contrary to previous conflicting reports.
Introduction
- The study aims to thoroughly explore and compare the “luteinizing hormone” (LH) and “follicle-stimulating hormone” (FSH) activities of the horse (Equus caballus) and the donkey (Equus asinus).
- Prior research indicated that horse LH and chorionic gonadotrophin (CG) only express LH activity in equids, but present dual LH and FSH activities in other species.
- Donkey LH and CG, though sharing identical peptide sequences, are less understood. Previous literature shows mixed results regarding their FSH activity in non-donkey species.
Methods
- The hormones were expressed in recombinant form using COS-7 cells, a type of monkey kidney cell line commonly used in biology labs to propagate recombinant DNA to produce required substances.
- The LH activities were tested using a rat Leydig cell bioassay, while the FSH activities were estimated through a bioassay using Y1 cells, which are engineered to express the human FSH receptor.
- Additionally, Human chorionic gonadotrophin (hCG) was similarly expressed and analysed in the same system for comparison.
Results
- The results indicated that recombinant donkey LH was almost twice as active as horse LH in the LH bioassay. However, its activity was much lower in the FSH bioassay—about five times less active than horse LH.
- Human CG, on the other hand, was about three times less active than horse LH/CG in the LH bioassay but showed zero activity in the FSH bioassay.
Conclusion
- The research concludes that donkey LH/CG shows significant FSH activity in heterologous species, contradicting the idea that its FSH activity could be due to contamination.
- These findings add new insights into the cross-species hormone activity and provide a better understanding of horse and donkey reproductive biology.
Cite This Article
APA
Chopineau M, Martinat N, Troispoux C, Marichatou H, Combarnous Y, Stewart F, Guillou F.
(1997).
Expression of horse and donkey LH in COS-7 cells: evidence for low FSH activity in donkey LH compared with horse LH.
J Endocrinol, 152(3), 371-377.
https://doi.org/10.1677/joe.0.1520371 Publication
Researcher Affiliations
- Station de Physiologie de la Reproduction, Institut National de la Recherche Agronomique, URA INRA/CNRS 1291, Nouzilly, France.
MeSH Terms
- Animals
- Biological Assay
- Biological Availability
- COS Cells
- Equidae / metabolism
- Follicle Stimulating Hormone / metabolism
- Gonadotropins, Equine / metabolism
- Horses / metabolism
- Luteinizing Hormone / biosynthesis
- Luteinizing Hormone / metabolism
- Recombinant Proteins / biosynthesis
- Recombinant Proteins / metabolism
Citations
This article has been cited 3 times.- Min KS, Park JJ, Lee SY, Byambaragchaa M, Kang MH. Comparative gene expression profiling of mouse ovaries upon stimulation with natural equine chorionic gonadotropin (N-eCG) and tethered recombinant-eCG (R-eCG).. BMC Biotechnol 2020 Nov 11;20(1):59.
- Min KS, Park JJ, Byambaragchaa M, Kang MH. Characterization of tethered equine chorionic gonadotropin and its deglycosylated mutants by ovulation stimulation in mice.. BMC Biotechnol 2019 Aug 13;19(1):60.
- Sonawani A, Niazi S, Idicula-Thomas S. In silico study on binding specificity of gonadotropins and their receptors: design of a novel and selective peptidomimetic for human follicle stimulating hormone receptor.. PLoS One 2013;8(5):e64475.
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