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Doctoral Thesis
DOI
https://doi.org/10.11606/T.10.2017.tde-21032017-151159
Document
Author
Full name
Moana Rodrigues França
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Pirassununga, 2016
Supervisor
Committee
Binelli, Mario (President)
Milazzotto, Marcella Pécora
Perecin, Felipe
Seneda, Marcelo Marcondes
Watanabe, Yeda Fumie
Title in Portuguese
Perfis endócrinos peri-ovulatórios influenciam o transporte, metabolismo e disponibilidade de aminoácidos no lúmen uterino de vacas de corte durante o diestro inicial
Keywords in Portuguese
Aminoácidos
Bovino
Esteroides sexuais
Proteínas carreadoras de soluto
Receptividade uterina
Abstract in Portuguese
Em vacas de corte, folículos pré-ovulatórios (FPO) maiores, maiores concentrações de estradiol (E2) no proestro/estro e progesterona (P4) no diestro favorecem o crescimento do concepto e a fertilidade. Porém, os mecanismos mediados pelos esteroides femininos que influenciam a receptividade uterina ao embrião precisam ser esclarecidos. Os aminoácidos (AA) são componentes das secreções uterinas que são cruciais para a sobrevivência do embrião antes da implantação. A hipótese deste trabalho é que o tamanho do FPO e as concentrações de E2 e P4 modulam a abundância de transcritos relacionados ao transporte e metabolismo de AA no endométrio e afetam a concentração luminal de AA. Para isso, o crescimento folicular de vacas Nelore foi manipulado com o objetivo de formar dois grupos: FPO grande e CL grande (FG-CLG) e FPO pequeno e CL pequeno (FP-CLP). No Dia 4 (D4; Exp 1) e Dia 7 (D7; Exp 2) após a injeção de GnRH para indução da ovulação, foram coletados tecido endometrial e lavado uterino post-mortem. A abundância de transcritos foi avaliada por qRT-PCR e as concentrações de AA nos lavados foram quantificadas por HPLC. No Exp 1, o tamanho do FPO, concentrações plasmáticas de E2 no D-1 e de P4 no D4 foram 15,70mm±0,43 vs. 11,31mm±0,23 (p<0,01), 2,44pg/ml±0,19 vs. 0,65pg/ml (p<0,01) e 1,40ng/ml±0,23 vs. 0,80ng/ml±0,10 (p<0,01) para os grupos FG-CLG vs. FP-CLP, respectivamente. No Exp 2, o tamanho do FPO, concentrações plasmáticas de E2 no D-1 e de P4 no D7 foram 13,18mm±0,44 vs. 10,63mm±0,30 (p<0,01), 2,30pg/ml±0,57 vs. 0,50pg/ml±0,13 (p<0,01) e 3,68ng/ml±0,38 vs. 2,49ng/ml±0,43 (p=0,04) para os grupos FG-CLG vs. FP-CLP, respectivamente. No D4 a abundância de SLC1A4, SLC38A1, SLC6A6, SLC7A4 e SLCY e no D7, SLC1A4, SLC6A1, SLC6A14, SLC7A4, SLC7A8, SLC38A1, SLC38A7, SLC43A2 e DDO foi maior no endométrio dos animais do grupo FG-CLG (p<0,05). No D4, maiores concentrações de taurina, alanina e ácido α-aminobutírico foram observadas no grupo FP-CLP (p<0,05). Em contraste, menores concentrações de valina e cistationina foram encontradas nos lavados uterinos do D7 dos animais do grupo FP-CLP (p<0,05). No D4, os animais do grupo FG-CLG, associado a maior fertilidade, apresentaram menor quantidade de AA nas secreções uterinas, porém, a abundância dos transportadores de AA foi compatível com maior transporte em comparação aos animais do grupo FP-CLP. Esses resultados sugerem que antes do embrião se mover do oviduto ao útero, o transporte e metabolismo dos AA prioriza a preparação das células endometriais para receber o embrião e não o acúmulo nas secreções uterinas. Porém, no D7, quando o embrião está em contato direto com as secreções uterinas, os genes relacionados ao transporte de AA no endométrio e a concentração de AA no histotrofo são estimulados nas vacas do grupo FG-CLG. Portanto o metabolismo e transporte de AA no sentido das células endometriais ou das secreções uterinas pode ser um mecanismo importante para a receptividade materna.
Title in English
Pre-ovulatory endocrine profiles influence endometrial amino acids transport, metabolism and availability in the uterine lumen during early diestrus in beef cows
Keywords in English
Amino acids
Cattle
Sex steroids
Solute Carrier Proteins
Uterine receptivity
Abstract in English
In beef cattle, a large size of the pre-ovulatory follicle (POF) and resulting elevated proestrus/estrus estradiol (E2) and diestrus progesterone (P4) concentrations positively affect conceptus growth and fertility. However, sex-steroid-mediated mechanisms that influence uterine receptivity to the embryo need to be elucidated. Amino acids are important components of maternally-derived secretions that are crucial for embryo survival before implantation. The hypothesis is that the size of the POF, E2 and P4 concentrations modulate endometrial abundance of solute carrier proteins (SLC) transcripts related to AA transport and metabolism and subsequently affect lumenal amino acids concentrations. Therefore, follicle growth of Nelore cows was manipulated to produce two experimental groups: large POF and CL (LF-LCL group) and small POF and CL (SF-SCL group). On Day 4 (D4; Experiment 1) and Day 7 (D7; Experiment 2) post GnRH injection to induce ovulation, endometrial tissue and uterine washings were collected post-mortem. Transcript abundance was evaluated by qRT-PCR and amino acid concentrations were quantified in washings by HPLC. On Experiment 1, POF size, plasma E2 concentration on D-1, and plasma concentration of P4 on D4 were 15.70mm±0.43 vs. 11.31mm±0.23 (p<0.01), 2.44pg/ml±0.19 vs. 0.65pg/ml (p<0.01) and 1.40ng/ml±0.23 vs. 0.80ng/ml±0.10 (p<0.01) for the LF-LCL vs. SF-SCL groups, respectively. For Experiment 2, POF size, plasma E2 concentration on D-1 and plasma P4 concentration on D7 were 13.18mm±0.44 vs. 10.63mm±0.30 (p<0.01), 2.30pg/ml±0.57 vs. 0.50pg/ml±0.13 (p<0.01) and 3.68ng/ml±0.38 vs. 2.49ng/ml±0.43 (p=0.04) for the LF-LCL vs. SF-SCL groups, respectively. On D4, abundance of SLC6A6, SLC7A4, SLC17A5, SLC38A1, SLC38A7 and SLCY and on D7, SLC1A4, SLC6A1, SLC6A14, SLC7A4, SLC7A7, SLC7A8, SLC17A5, SLC38A1, SLC38A7, SLC43A2 and DDO was greater in the endometrium of cows from the LF-LCL group (p<0.05). On D4, higher concentrations of taurine, alanine and α-aminobutiric acid were observed in SF-SCL (p<0.05). In contrast, lower concentrations of valine and cystathionine were quantified in D7 uterine washings from SF-SCL cows (p<0.05). On D4, animals from LF-LCL group, associated with greater fertility, presented less amino acid content in uterine secretion but abundance of transporters was compatible to greater transport in comparison to animals from SF-SCL group. This suggests that before embryo moves from oviduct to uterus, amino acids transport and metabolism pathways prioritizes endometrium cells preparation for receiving the embryo but not accumulation in uterine secretions. However, on D7, when the embryo is in direct contact with uterine secretions, genes related to amino acids transport in endometrium and amino acids concentration in histotroph are up-regulated in LF-LCL cows. The latter insights indicate that amino acids metabolism and transport, towards endometrial cells or uterine secretions, might be mechanisms contributing to maternal receptivity.
 
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Publishing Date
2017-07-17
 
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