Differential expression of LHCGR and its isoforms is associated to the variability in superovulation responses of Gir cattle

The aim of this study was to evaluate the pattern of expression of LHCGR isoforms in Gir heifers characterized as good (10.3 ± 1.2 ova/embryos per flush, n = 5) or poor responders (1.1 ± 0.3 ova/embryos per flush, n = 5) to superovulation protocols. In both groups, an adapted ultrasound-guided folli...

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Bibliographic Details
Published in:Theriogenology 2019-03, Vol.126, p.68-74
Main Authors: Wohlres-Viana, S., Arashiro, E.K.N., Minare, T.P., Fernandes, C.A.C., Grazia, J.G.V., Siqueira, L.G.B., Machado, M.A., Viana, J.H.M.
Format: Article
Language:English
Subjects:
Embryo transfer
In vivo embryo production
Luteinizing hormone
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Summary:The aim of this study was to evaluate the pattern of expression of LHCGR isoforms in Gir heifers characterized as good (10.3 ± 1.2 ova/embryos per flush, n = 5) or poor responders (1.1 ± 0.3 ova/embryos per flush, n = 5) to superovulation protocols. In both groups, an adapted ultrasound-guided follicular aspiration system was used to collect granulosa cells from 8 mm follicles formed either during a synchronized, non-stimulated follicular wave (no stimulation control, NS) or on the fourth day of a superovulation protocol (SOV) induced with 200 IU of pFSH. The recovered follicular fluid was centrifuged and granulosa cells were washed with NaCl 0.9% and kept in RNAlater®. RNA extraction was performed using a commercial RNeasy Micro Kit and eluted samples were quantified and reverse transcribed using the commercial Superscript III kit. cDNA samples were amplified by real-time PCR using a primer to target LH/hCG receptor gene - not selective for LHCGR isoforms (total LHCGR) - and four sets of isoforms selective primers (S1, S10, S10 + 11, and S11). Analyses were performed using the REST software and expression levels are shown as mean ± SEM. Under physiological conditions (NS), poor responders had a higher expression of total LHCGR (4.9 ± 1.7 fold-change, P 
ISSN:0093-691X
1879-3231
DOI:10.1016/j.theriogenology.2018.12.004