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1.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201586

RESUMO

The molecular mechanism underlying embryonic implantation is vital to understand the correct communications between endometrium and developing conceptus during early stages of pregnancy. This study's objective was to determine molecular changes in the uterine endometrial proteome during the preimplantation and peri-implantation between 9 days (9D), 12 days (12D), and 16 days (16D) of pregnant Polish Large White (PLW) gilts. 2DE-MALDI-TOF/TOF and ClueGOTM approaches were employed to analyse the biological networks and molecular changes in porcine endometrial proteome during maternal recognition of pregnancy. A total of sixteen differentially expressed proteins (DEPs) were identified using 2-DE gels and MALDI-TOF/TOF mass spectrometry. Comparison between 9D and 12D of pregnancy identified APOA1, CAPZB, LDHB, CCT5, ANXA4, CFB, TTR upregulated DEPs, and ANXA5, SMS downregulated DEPs. Comparison between 9D and 16D of pregnancy identified HP, APOA1, ACTB, CCT5, ANXA4, CFB upregulated DEPs and ANXA5, SMS, LDHB, ACTR3, HP, ENO3, OAT downregulated DEPs. However, a comparison between 12D and 16D of pregnancy identified HP, ACTB upregulated DEPs, and CRYM, ANXA4, ANXA5, CAPZB, LDHB, ACTR3, CCT5, ENO3, OAT, TTR down-regulated DEPs. Outcomes of this study revealed key proteins and their interactions with metabolic pathways involved in the recognition and establishment of early pregnancy in PLW gilts.


Assuntos
Implantação do Embrião/fisiologia , Endométrio/metabolismo , Prenhez/metabolismo , Proteínas/metabolismo , Animais , Feminino , Gravidez , Proteínas/análise , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Suínos
2.
Int J Mol Sci ; 22(7)2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33915762

RESUMO

The corpus luteum (CL) is a temporary endocrine gland vital for pregnancy establishment and maintenance. Estradiol-17ß (E2) is the major embryonic signal in pigs supporting the CL's function. The mechanisms of the luteoprotective action of E2 are still unclear. The present study aimed to determine the effect of E2 on luteal expression of factors involved in CL function. An in vivo model of intrauterine E2 infusions was applied. Gilts on day 12 of pregnancy and the estrous cycle were used as referential groups. Concentrations of E2 and progesterone were elevated in CLs of gilts receiving E2 infusions, compared to placebo-treated gilts. Estradiol-17ß stimulated luteal expression of DNA-methyltransferase 1 (DNMT1), but decreased expression of DNMT3B gene and protein, as well as DNMT3A protein. Similar results for DNMT3A and 3B were observed in CLs on day 12 of pregnancy compared to day 12 of the estrous cycle. Intrauterine infusions of E2 altered luteal expression of the genes involved in CL function: PTGFR, PTGES, STAR, HSD17B1, CYP19A1, and PGRMC1. Our findings indicate a role for E2 in expression regulation of factors related to CL function and a novel potential for E2 to regulate DNA methylation as putative physiological mechanisms controlling luteal gene expression.


Assuntos
Corpo Lúteo/enzimologia , DNA (Citosina-5-)-Metiltransferases/metabolismo , Estradiol/fisiologia , Prenhez/metabolismo , Animais , Estradiol/administração & dosagem , Feminino , Gravidez , Progesterona/metabolismo , Suínos
3.
Reprod Domest Anim ; 56(5): 713-724, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33547667

RESUMO

MiRNAs-containing extracellular vesicles (EVs) possess the unique function of mediating intercellular communication and participating in many biological processes such as post-transcriptional gene regulation of embryo implantation and placental development. In the present study, Illumina small-RNA sequencing was used to identify differentially expressed (DE) miRNAs in serum EVs of pregnant (P) and non-pregnant (NP) Kazakh sheep at Day 17 from mating. The specifically and differentially expressed miRNAs at early pregnancy in sheep were verified by using RT-PCR. The target genes of DE miRNAs were predicted by bioinformatics software, and the functional and pathway enrichment analysis was performed on Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) terms. A total of 562 miRNAs (210 novel miRNAs) were identified by sequencing, of which 57 miRNAs were differentially expressed, 49 were up-regulated, 8 were down-regulated and 22 novel miRNAs were specifically expressed in the pregnant sheep. Eight highly expressed known miRNA (miR-378-3p, miR-320-3p, miR-22-3p, let-7b, miR-423-3p, miR-221, miR-296-3p, miR-147-3p) in pregnant group were down-regulated in the control group. miRNAs-containing pregnancy-related terms and regulatory pathways regulation were enriched using both GO and KEGG analyses. Moreover, we also envisioned a miRNA-mRNA interaction network to understand the function of miRNAs involved in the early pregnancy serum regulatory network. The results of RT-PCR verification confirmed the reliability of small-RNA sequencing. Among them, miR-22-3p and miR-378-3p were significantly differentially expressed (DE) between pregnant sheep and non-pregnant group (p <  0.01). The site at which oar-miR-22-3p binds MAPK3 was determined with a dual-luciferase system. This is the first integrated analysis of the expression profiles of EV-miRNAs and their targets during early pregnancy in ewes. These data identify key miRNAs that influence the implantation of sheep in the early stage of pregnancy, and provide theoretical basis for further molecular regulatory mechanisms research.


Assuntos
Vesículas Extracelulares/metabolismo , MicroRNAs/metabolismo , Prenhez/sangue , Animais , Vesículas Extracelulares/genética , Feminino , Perfilação da Expressão Gênica/veterinária , MicroRNAs/genética , Gravidez , Prenhez/metabolismo , Carneiro Doméstico
4.
J Dairy Sci ; 104(1): 1053-1072, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33189277

RESUMO

The transition from pregnancy to lactation is the most challenging period for high-producing dairy cows. The liver plays a key role in biological adaptation during the peripartum. Prior works have demonstrated that hepatic glucose synthesis, cholesterol metabolism, lipogenesis, and inflammatory response are increased or activated during the peripartum in dairy cows; however, those works were limited by a low number of animals used or by the use of microarray technology, or both. To overcome such limitations, an RNA sequencing analysis was performed on liver biopsies from 20 Holstein cows at 7 ± 5d before (Pre-P) and 16 ± 2d after calving (Post-P). We found 1,475 upregulated and 1,199 downregulated differently expressed genes (DEG) with a false discovery rate adjusted P-value < 0.01 between Pre-P and Post-P. Bioinformatic analysis revealed an activation of the metabolism, especially lipid, glucose, and amino acid metabolism, with increased importance of the mitochondria and a key role of several signaling pathways, chiefly peroxisome proliferators-activated receptor (PPAR) and adipocytokines signaling. Fatty acid oxidation and gluconeogenesis, with a likely increase in amino acid utilization to produce glucose, were among the most important functions revealed by the transcriptomic adaptation to lactation in the liver. Although gluconeogenesis was induced, data indicated decrease in expression of glucose transporters. The analysis also revealed high activation of cell proliferation but inhibition of xenobiotic metabolism, likely due to the liver response to inflammatory-like conditions. Co-expression network analysis disclosed a tight connection and coordination among genes driving biological processes associated with protein synthesis, energy and lipid metabolism, and cell proliferation. Our data confirmed the importance of metabolic adaptation to lipid and glucose metabolism in the liver of early Post-P cows, with a pivotal role of PPAR and adipocytokines.


Assuntos
Bovinos/metabolismo , Fígado/metabolismo , Prenhez/metabolismo , Transcriptoma , Adaptação Fisiológica , Animais , Biologia Computacional , Feminino , Gluconeogênese , Lactação , Metabolismo dos Lipídeos/fisiologia , Período Periparto , Período Pós-Parto/metabolismo , Gravidez , Regulação para Cima
5.
Anim Sci J ; 91(1): e13494, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33368854

RESUMO

Assessing the resilience of Awassi sheep to water shortages during pregnancy, thereby investigating the effects of water scarcity on gestation efficiency in animals, could provide valuable and pertinent insight into future scenarios posed by climate change risks. In this study, 40 pregnant Awassi ewes randomly allocated to free watering group (C, n = 20) received water ad libitum and the water restricted group (WR) received 50% less water than the amount provided to group C. Water restriction decreased (p < .05) lambs' birth weights, placental weights (PWs), and cotyledon numbers (CNs). Placental efficiency (PE) and cotyledon efficiency (CE) were significantly higher in the WR group (p < .05). A marked difference in cotyledon weight - an increase of 12.1% - was recorded in the C group. The body weights of the pregnant ewes in the WR group decreased significantly (p < .05) by 22% during pregnancy. Significant increases (p < .05) in plasma ADH, cholesterol, Cl- , and Na+ levels were observed in the WR group ewes, indicating intense dehydration. We conclude that the Awassi breed of sheep can endure 50% water restriction during pregnancy and maintain successful parturition, a key outcome for sustainability.


Assuntos
Secas , Placenta/fisiologia , Prenhez/metabolismo , Prenhez/fisiologia , Ovinos/metabolismo , Ovinos/fisiologia , Privação de Água/fisiologia , Animais , Peso ao Nascer/fisiologia , Cloro/sangue , Colesterol/sangue , Mudança Climática , Feminino , Tamanho do Órgão , Placenta/anatomia & histologia , Gravidez , Sódio/sangue , Vasopressinas/sangue
6.
Sci Rep ; 10(1): 20798, 2020 11 27.
Artigo em Inglês | MEDLINE | ID: mdl-33247230

RESUMO

Supplementation of cattle diets with n-3-polyunsaturated fatty acids (PUFA) can improve reproductive efficiency. Conversely, short-term fluctuations in feed supply can impact pregnancy establishment. The objectives of this study were to examine the effects of (1) dietary supplementation with n-3-PUFA and (2) post-insemination plane of nutrition on the endometrial transcriptome. Beef crossbred heifers were offered concentrate based diets fortified with n-3-PUFA (PUFA; n = 32) or not (CONT; n = 28) for 30 days prior to breeding at a synchronised oestrous. Following artificial insemination, heifers were allocated within treatment to either a high or low plane of nutrition. Heifers were maintained on these diets for 16 days following which endometrial tissue was harvested at slaughter for subsequent RNAseq analysis. The influence of pregnancy status on the endomentrial transcriptome, within each dietary treatment group, was also examined. Post-insemination diet affected (P < 0.05) the endometrial transcriptome. Specifically, within n-3-PUFA-supplemented heifers, genes involved in embryonic development and mTOR signalling pathways, important in pregnancy establishment, were identified as differentially expressed. Results indicate that dietary supplementation of cattle diets with n-3-PUFA may have a positive effect on the expression of key fertility-related genes and pathways, during the critical window of maternal recognition of pregnancy, particularly where animals are underfed.


Assuntos
Bovinos/genética , Bovinos/metabolismo , Dieta/veterinária , Suplementos Nutricionais , Endométrio/metabolismo , Ácidos Graxos Ômega-3/administração & dosagem , Prenhez/genética , Prenhez/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Proliferação de Células/genética , Desenvolvimento Embrionário/genética , Feminino , Redes Reguladoras de Genes , Inseminação Artificial/veterinária , Estado Nutricional , Gravidez , Transdução de Sinais/genética , Transcriptoma
7.
BMC Genomics ; 21(1): 626, 2020 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-32917128

RESUMO

BACKGROUND: The environmental impact of pig farming need to be reduced, with phosphorus (P) being of particular interest. Specified dietary regimens and management systems contribute to meet environmental concerns and reduce economic constrains. However, pregnant and lactating sows represent vulnerable individuals, whose reproductive potential and metabolic health status relies on adequate supply of macro- and micronutrients. The aim of this study was to investigate, whether sows fed with a dietary P content that is below or above current recommendations are capable to maintain mineral homeostasis during the reproduction cycle and which endogenous mechanisms are retrieved therefore in kidney and jejunum. Nulliparous gilts were fed iso-energetic diets with recommended (M), reduced (L), or high (H) amounts of mineral P supplements throughout gestation and lactation periods. Blood metabolites and hormones referring to the P homeostasis were retrieved prior to term (110 days of gestation) and at weaning (28 days of lactation). Transcriptional responses in kidney cortex and jejunal mucosa were analyzed using RNA sequencing. RESULTS: The variable dietary P content neither led to an aberration on fertility traits such as total weaned piglets nor to an effect on the weight pattern throughout gestation and lactation. Serum parameters revealed a maintained P homeostasis as reflected by unaltered inorganic P and calcium levels in L and H fed groups. The serum calcitriol levels were increased in lactating L sows. The endocrine responses to the dietary challenge were reflected at the transcriptional level. L diets led to an increase in CYP27B1 expression in the kidney compared to the H group and to an altered gene expression associated with lipid metabolism in the kidney and immune response in the jejunum. CONCLUSIONS: Our results suggest that current P requirements for gestating and lactating sows are sufficient and over supplementation of mineral P is not required. Shifts in renal and jejunal expression patterns between L and H groups indicate an affected intermediate metabolism, which long-term relevance needs to be further clarified.


Assuntos
Jejuno/metabolismo , Rim/metabolismo , Fósforo na Dieta/metabolismo , Prenhez/metabolismo , Suínos/metabolismo , Transcriptoma , Adaptação Fisiológica , Ração Animal/normas , Animais , Feminino , Lactação/metabolismo , Fósforo na Dieta/normas , Gravidez , Suínos/genética , Suínos/fisiologia
8.
Anim Reprod Sci ; 219: 106532, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32828407

RESUMO

Adipokines such as chemerin affect metabolic status and reproductive function in many species. The hypothesis in the present study was that there were chemerin mRNA transcript and protein in the pituitary of pigs and that relative abundances fluctuate during the estrous cycle and early pregnancy. Chemerin is thought to modulate luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion during the estrous cycle. Changes in the relative abundance of chemerin mRNA transcript and protein in anterior (AP) and posterior (PP) pituitaries of pigs were investigated, for the first time in the present study, during four phases of the estrous cycle and four periods of early pregnancy. Chemerin protein was localized in gonadotrophs, thyrotrophs and somatotrophs during the estrous cycle and early gestation. Chemerin treatments affected both basal, GnRH- and/or insulin-induced LH and FSH production, with there being variations with phase of the estrous cycle when tissues were collected. These findings indicate chemerin may be produced locally in the pituitary and may affect female reproductive function by controlling the release of LH and FSH from AP cells.


Assuntos
Quimiocinas , Ciclo Estral , Hipófise/metabolismo , Prenhez , Suínos , Animais , Células Cultivadas , Quimiocinas/genética , Quimiocinas/metabolismo , Ciclo Estral/genética , Ciclo Estral/metabolismo , Feminino , Hormônio Foliculoestimulante/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Hormônio Luteinizante/metabolismo , Gravidez , Prenhez/genética , Prenhez/metabolismo , RNA Mensageiro/metabolismo , Suínos/genética , Suínos/metabolismo
9.
Anim Reprod Sci ; 218: 106505, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32507267

RESUMO

This study was conducted to evaluate the changing concentrations of metabolic hormones and metabolites in pregnant (P) and pseudopregnant (PP) rabbit does. Twenty-five New Zealand White rabbit does were submitted to artificial insemination (AI) and then classified as P (n = 15) or PP (n = 10). Blood samples were collected weekly until day 32 post AI. During pregnancy, leptin concentrations were greater on Days 14 and 21 (P < 0.05), while insulin was greater on days 21 and 32 post AI (P < 0.05) compared to PP does. The triiodothyronine/thyroxine (T3/T4) ratio was greater in the first and last week (P < 0.001); whereas, cortisol concentrations were greater in the last week of pregnancy and after parturition (P < 0.01) compared with that of PP does. Non-esterified fatty acids (NEFA) concentrations increased from day 7 until day 32 post AI (P < 0.05). Glucose concentrations were unchanged throughout pregnancy although concentrations were positively associated with litter size. These results indicate concentrations of hormones and metabolites change during pregnancy to ensure energy requirements are met for both the foetuses and the maternal tissues. Physiological hyperleptinemia, hyperinsulinemia, and changes in cortisol as well as thyroid hormones indicate there is an adaptation of metabolic functions induced by pregnancy. These adaptations could be mediated by gonadal steroids because changes mainly occur in the second half of pregnancy when the profile of the sex hormones differs between P and PP does.


Assuntos
Metabolismo Energético/fisiologia , Homeostase/fisiologia , Prenhez , Pseudogravidez/veterinária , Coelhos/fisiologia , Adaptação Fisiológica , Animais , Feminino , Hidrocortisona/sangue , Insulina/sangue , Resistência à Insulina , Leptina/sangue , Gravidez , Prenhez/metabolismo , Pseudogravidez/metabolismo , Tri-Iodotironina/sangue
10.
Anim Sci J ; 91(1): e13378, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32329195

RESUMO

Interferon-tau (IFNT) regulates maternal recognition during early pregnancy in ruminants. The liver can serve as a hematopoietic organ, and it has immune functions. This study hypothesized whether mRNA and proteins of interferon-stimulated genes (ISGs) induced by early pregnancy are upregulated in maternal liver. Therefore, we determined the expression of interferon-stimulated gene 15-kDa protein (ISG15), 2',5'-oligoadenylate synthetase 1 (OAS1), myxovirus resistance protein 1 (MX1), interferon-gamma-inducible protein 10 (IP-10), and signal transducer and activator of transcription 1 (STAT1) in maternal livers during early pregnancy in sheep. Ovine livers were sampled on day 16 of the estrous cycle, and days 13, 16, and 25 of pregnancy, and expression of ISGs was detected by quantitative real-time PCR, Western blot, and immunohistochemistry analysis. Our results showed that there were increases in expression of the mRNA and proteins of ISG15, OAS1, IP-10, STAT1, and MX1 during early pregnancy. STAT1 protein was limited to the hepatocytes, and endothelial cells of proper hepatic arteries and hepatic portal veins. In conclusion, the upregulation of ISG15, OAS1, IP-10, STAT1, and MX1 proteins may be implicated in maternal hepatic immune adjustment and other functions during early pregnancy in sheep.


Assuntos
2',5'-Oligoadenilato Sintetase/genética , 2',5'-Oligoadenilato Sintetase/metabolismo , Quimiocina CXCL10/genética , Quimiocina CXCL10/metabolismo , Citocinas/genética , Citocinas/metabolismo , Expressão Gênica , Fígado/metabolismo , Proteínas de Resistência a Myxovirus/genética , Proteínas de Resistência a Myxovirus/metabolismo , Prenhez/metabolismo , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Ovinos/genética , Ovinos/fisiologia , Ubiquitinas/genética , Ubiquitinas/metabolismo , Animais , Ciclo Estral/genética , Ciclo Estral/metabolismo , Feminino , Fígado/imunologia , Gravidez , Prenhez/imunologia , Ovinos/imunologia , Regulação para Cima
11.
Sci Rep ; 10(1): 2089, 2020 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-32034259

RESUMO

Despite their importance in mammalian reproduction, substances in the oxytocin-prostaglandins pathways have not been investigated in the horse placenta during most of pregnancy and parturition. Therefore, we quantified placental content of oxytocin (OXT), oxytocin receptor (OXTR), and prostaglandin E2 and F2 alpha during days 90-240 of pregnancy (PREG), physiological parturition (PHYS), and parturition with fetal membrane retention (FMR) in heavy draft horses (PREG = 13, PHYS = 11, FMR = 10). We also quantified OXTR and prostaglandin endoperoxide synthase-2 (PTGS2) mRNA expression and determined the immunolocalization of OXT, OXTR, and PTGS2. For relative quantification of OXT and OXTR, we used western blotting with densitometry. To quantify the prostaglandins, we used enzyme immunoassays. For relative quantification of OXTR and PTGS2, we used RT-qPCR. For immunolocalization of OXT, OXTR, and PTGS2, we used immunohistochemistry. We found that OXT was present in cells of the allantochorion and endometrium in all groups. PTGS2 expression in the allantochorion was 14.7-fold lower in FMR than in PHYS (p = 0.007). These results suggest that OXT is synthesized in the horse placenta. As PTGS2 synthesis is induced by inflammation, they also suggest that FMR in heavy draft horses may be associated with dysregulation of inflammatory processes.


Assuntos
Membranas Extraembrionárias/metabolismo , Cavalos/fisiologia , Ocitocina/metabolismo , Parto/metabolismo , Placenta/metabolismo , Prenhez/metabolismo , Prostaglandinas/metabolismo , Animais , Membranas Extraembrionárias/fisiologia , Feminino , Cavalos/metabolismo , Redes e Vias Metabólicas , Ocitocina/fisiologia , Parto/fisiologia , Placenta/fisiologia , Gravidez , Prenhez/fisiologia , Prostaglandinas/fisiologia
12.
Nutrients ; 12(2)2020 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-32098129

RESUMO

The leucine metabolite, ß-hydroxy-ß-methyl butyrate (HMB), is widely used in human nutrition and animal production as a nutritional supplement. Although the HMB usage during late gestation has been demonstrated to have a positive effect on fetal development, knowledge on net absorption and metabolism of HMB and impact of HMB on branched chain amino acids (BCAAs) metabolism is lacking. To address this, we conducted a study using pigs during the perinatal period as a model organism. Eight-second parity sows were fitted with indwelling catheters in the femoral artery and in the portal, hepatic, femoral, and mesenteric veins. Eight hourly sets of blood samples were taken starting 30 min before the morning meal on day -10 and day -3 relative to parturition. Four control (CON) sows were fed a standard lactation diet from day -15 and throughout the experiment, and 4 HMB sows were fed the control diet supplemented with 15 mg Ca(HMB)2/kg body weight mixed in one third of the morning meal from day -10 until parturition. Blood gases, plasma metabolites, milk compositions, and apparent total tract digestibility of nutrients were measured. Arterial plasma concentrations of HMB (p < 0.001), Cys (p < 0.001), and Lys (p < 0.10) were increased in HMB supplemented sows, while arterial plasma triglycerides concentration was decreased (p < 0.05). The net portal recovery of Ala and Asp were increased in HMB sows (p < 0.05). Sows fed HMB had increased hepatic vein flow and net hepatic fluxes of Met, Asn, and Gln (p < 0.05). In contrast, the femoral extraction rates of Ala and Ser were decreased by dietary HMB supplementation (p < 0.05). Dietary HMB treatment and sampling time relative to feeding had an interaction on arterial concentrations, net portal fluxes, and femoral extraction rates of BCAAs. The net portal recovery of HMB was 88%, while 14% of supplemented HMB was excreted through urine and 4% through feces. Moreover, the gastrointestinal tract metabolized 8% while the liver metabolized 12%. Finally, 26% of the daily intake of HMB was secreted via colostrum at the day of farrowing. This study demonstrated that dietary HMB supplementation increased net uptake of amino acids and increased fatty acid oxidation through improving blood flow and insulin sensitivity during the late gestation. Most importantly, oral HMB administration could maintain a stable postprandial absorption and altered metabolism in BCAAs. Net portal flux of HMB at 5.5 to 6.5 h after feeding approached zero, indicating that HMB ideally should be administrated two or three times, daily.


Assuntos
Aminoácidos de Cadeia Ramificada/metabolismo , Suplementos Nutricionais , Prenhez/metabolismo , Valeratos/metabolismo , Aminoácidos de Cadeia Ramificada/sangue , Ração Animal , Animais , Ácidos Graxos/sangue , Ácidos Graxos/metabolismo , Feminino , Absorção Gastrointestinal/fisiologia , Insulina/sangue , Insulina/metabolismo , Resistência à Insulina/fisiologia , Modelos Animais , Oxirredução , Gravidez , Prenhez/sangue , Suínos , Valeratos/administração & dosagem , Valeratos/sangue
13.
Reproduction ; 159(3): 303-314, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31990672

RESUMO

Decidualization is a critical process for successful embryo implantation and subsequent placenta formation. The characterization and physiological function of lncRNA during decidualization remain largely unknown. In the present study, we conducted RNA-sequencing analysis to compare gene expression between decidua of days 6 and 8, and normal pregnant endometrium (day 4). A total of 2332 high-confidence putative lncRNA transcripts were expressed. Functional clustering analysis of cis and trans lncRNA targets showed that differentially expressed lncRNAs may regulate multiple gene ontology terms and pathways that have important functions in decidualization. Subsequent analyses using qRT-PCR validated that eight of all lncRNAs were differentially regulated in mice uteri during decidualization, both in vivo and in vitro. Furthermore, we showed that differentially expressed lncRNA of Hand2os1 was specifically detected in stromal cells on days 2 to 5 of pregnancy and was strongly upregulated in decidual cells on days 6-8 of pregnancy. Similarly, Hand2os1 expression was also strongly expressed in decidualized cells following artificial decidualization, both in vivo and in vitro. In uterine stromal cells, P4 was able to significantly upregulate the expression of Hand2os1, but upregulation was impeded by RU486, whereas E2 appeared to have no regulating effect on Hand2os1 expression. Concurrently, Hand2os1 significantly promoted the decidual process in vitro and dramatically increased decidualization markers Prl8a2 and Prl3c1. Our results provide a valuable catalog for better understanding of the functional roles of lncRNAs in pregnant mouse uteri, as it relates to decidualization.


Assuntos
Endométrio/metabolismo , Prenhez/metabolismo , Progesterona/metabolismo , RNA Longo não Codificante/metabolismo , Receptores de Progesterona/metabolismo , Animais , Feminino , Perfilação da Expressão Gênica , Hormônios Esteroides Gonadais/metabolismo , Masculino , Camundongos , Gravidez , Pseudogravidez/metabolismo
14.
Reproduction ; 159(4): 493-501, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31967970

RESUMO

Maintenance of a suitable uterine milieu is important for embryo development and subsequent implantation during early pregnancy. High estrogen level in proestrous and estrous stages is essential for uterine anti-bacterial activity during preimplantation period. Lipocalin-2 is an essential molecule which prevents bacterial infection by sequestering iron. In this study, the highest expression of lipocalin-2 is observed in the endometrial epithelium on day 1 of normal pregnancy and pseudopregnancy, which exhibit a similar hormone scenario. By injecting the agonists for estrogen receptor α and estrogen receptor ß in ovariectomized mice, we found estrogen receptor α is the dominant member for estrogen regulation on lipocalin-2 expression. Estrogen treatment in estrogen receptor α-knockout mice further confirmed the role of estrogen receptor α. Using published data from whole-genome estrogen receptor α binding site assay, significant estrogen receptor α recruitment peaks are found at the downstream of lipocalin-2 gene after estrogen treatment. Furthermore, to study the anti-bacterial activity of lipocalin-2 in uterus, Escherichia coli is injected to mimic bacterial infection. Our results showed an obvious induction of lipocalin-2 in Escherichia coli-treated group. Taken together, this study indicates estrogen regulation of lipocalin-2 in uterine epithelium is mediated by estrogen receptor α, and lipocalin-2 may have anti-bacterial activity during early pregnancy.


Assuntos
Receptor alfa de Estrogênio/metabolismo , Lipocalina-2/metabolismo , Prenhez/metabolismo , Útero/metabolismo , Animais , Epitélio/metabolismo , Escherichia coli , Feminino , Lipopolissacarídeos , Camundongos , Camundongos Knockout , Gravidez , Pseudogravidez/metabolismo
15.
Mol Reprod Dev ; 87(1): 142-151, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31746519

RESUMO

Conceptus development and elongation is required for successful pregnancy establishment in ruminants and is coincident with the production of interferon τ (IFNT) and prostaglandins (PGs). In both the conceptus trophectoderm and endometrium, PGs are primarily synthesized through a prostaglandin-endoperoxide synthase 2 (PTGS2) pathway and modify endometrial gene expression and thus histotroph composition in the uterine lumen to promote conceptus growth and survival. Chemical inhibition of PG production by both the endometrium and the conceptus prevented elongation in sheep. However, the contributions of conceptus-derived PGs to preimplantation conceptus development remain unclear. In this study, CRISPR-Cas9 genome editing was used to inactivate PTGS2 in ovine embryos to determine the role of PTGS2-derived PGs in conceptus development and elongation. PTGS2 edited conceptuses produced fewer PGs, but secreted similar amounts of IFNT to their Cas9 control counterparts and elongated normally. Expression of PTGS1 was lower in PTGS2 edited conceptuses, but PPARG expression and IFNT secretion were unaffected. Content of PGs in the uterine lumen was similar as was gene expression in the endometrium of ewes who received either Cas9 control or PTGS2 edited conceptuses. These results support the idea that intrinsic PTGS2-derived PGs are not required for preimplantation embryo or conceptus survival and development in sheep.


Assuntos
Blastocisto/metabolismo , Ciclo-Oxigenase 2/metabolismo , Desenvolvimento Embrionário/genética , Prenhez/metabolismo , Ovinos/embriologia , Animais , Sistemas CRISPR-Cas , Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/genética , Transferência Embrionária/métodos , Endométrio/metabolismo , Feminino , Fertilização In Vitro/métodos , Edição de Genes , Expressão Gênica , Interferon Tipo I/biossíntese , PPAR gama/metabolismo , Gravidez , Proteínas da Gravidez/biossíntese , Prostaglandinas/biossíntese , Transdução de Sinais/genética
16.
Biol Reprod ; 102(2): 412-423, 2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-31504199

RESUMO

Circulating prostaglandin F2α metabolite (PGFM) after an oxytocin challenge was evaluated throughout the first 2 months of pregnancy in lactating Holstein cows. On day 11, 18, and 25 after artificial insemination (AI), and on days 32, 39, 46, 53, and 60 of pregnancy, cows were challenged with 50 IU oxytocin, i.m. Blood was collected before (0 min), 30, 60, 90, and 120 min after oxytocin for plasma PGFM concentrations. Ultrasound evaluations were performed for pregnancy diagnosis on day 32-60 post-AI. Nonpregnant (NP) cows on day 18 were designated by a lack of interferon-stimulated genes in peripheral blood leukocytes and Pregnant (P) based on day 32 ultrasound. On day 11, P and NP were similar with low PGFM and no effect of oxytocin on PGFM. On day 18, oxytocin increased PGFM (3-fold) in NP with little change in P cows. Comparing only P cows from day 11 to 60, basal circulating PGFM increased as pregnancy progressed, with day 11 and 18, lower than all days from day 25 to 60 of pregnancy. Oxytocin-induced PGFM in P cows on day 25 was greater than P cows on day 18 (2.9-fold). However, oxytocin-induced PGFM was lower on day 25 compared to day 53 and 60, with intermediate values on day 32, 39, and 46 of pregnancy. Thus, the corpus luteum (CL) of early pregnancy (day 11, 18) is maintained by suppression of PGF, as reflected by suppressed PGFM in this study. However, during the second month of pregnancy, uterine PGF secretion was not suppressed since basal PGFM and oxytocin-induced PGFM secretion were elevated. Apparently, mechanisms other than suppression of oxytocin receptors maintain CL after day 25 of pregnancy.


Assuntos
Corpo Lúteo/efeitos dos fármacos , Dinoprosta/análogos & derivados , Dinoprosta/metabolismo , Ocitocina/farmacologia , Prenhez/metabolismo , Animais , Bovinos , Corpo Lúteo/metabolismo , Dinoprosta/biossíntese , Feminino , Inseminação Artificial , Gravidez , Progesterona/sangue , Ultrassonografia
17.
Theriogenology ; 142: 196-206, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31606658

RESUMO

The maternal endometrium undergoes transformations during early pregnancy period to regulate the paracellular permeability across the epithelium and to enable adhesion between the trophoblast and endometrial epithelial cells. These transformations, under the influence of ovarian hormones, are associated with a partial loss in polarity of epithelial cell that is regulated by tight junctions (TJ), adherens junctions (AJ) and associated polarity protein complexes. This study examined the change in expression and distribution of proteins associated with TJs, AJs and apical partition defective (PAR) complex in porcine endometrium on Days 10, 13 and 16 of estrous cycle and pregnancy. Moreover, effect of hormones, progesterone (P4) and 17-ß estradiol (E2) on polar phenotype of endometrial epithelial cells was also investigated in vitro. There was pregnancy induced increase in gene and protein expression of TJ associated claudin-1 (CLDN1) on Day 13 of pregnancy as compared to corresponding day of estrous cycle and a decrease in TJ protein, zona occludens-1 (ZO-1) and PAR complex associated PAR6 expression levels on Day 16 of pregnancy (P < 0.05). Immunofluorescence studies revealed that on Days 10 and 13, TJ proteins occludin (OCLN) and ZO-1were primarily present in the apical region of lateral epithelial membrane. On Day 16 of pregnancy, whereas, OCLN redistributed into cytoplasm, ZO-1 decreased apically but was found to localize in the basal epithelium. The AJ proteins cadherin and ß-catenin were located at the apical epithelium on Day 10 of estrous cycle and pregnancy and Day 13 of estrous cycle. On Days 13 and 16 of pregnancy both proteins were expressed in the lateral membrane and co-localization between these proteins was observed on Day 16. On Day 10, PAR complex proteins PAR3, cell division control protein 42 (CDC42) and atypical protein kinase C (aPKC) ζ were observed in apical epithelium and in lateral membrane and CDC42 was also present in the cytoplasm of epithelium. Pregnancy induced redistribution of aPKCζ to cytoplasm and CDC42 to apical surface of luminal epithelium was observed on Days 13 and 16. The in vitro P4 and E2 treatment of epithelial cells mimicked in vivo results. These results indicate that P4 and E2 regulate alterations in epithelium that may facilitate embryo implantation and given the role of cadherin, catenin and CDC42 in embryo invasion, change in distribution of these proteins may limit the invasiveness of porcine conceptuses into the stroma.


Assuntos
Polaridade Celular/genética , Endométrio/metabolismo , Moléculas de Adesão Juncional/genética , Moléculas de Adesão Juncional/metabolismo , Prenhez , Suínos , Junções Aderentes/genética , Junções Aderentes/metabolismo , Animais , Células Cultivadas , Implantação do Embrião/genética , Feminino , Expressão Gênica , Idade Gestacional , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Gravidez , Prenhez/genética , Prenhez/metabolismo , Suínos/embriologia , Suínos/genética , Suínos/metabolismo , Junções Íntimas/genética , Junções Íntimas/metabolismo , Distribuição Tecidual
18.
Biol Reprod ; 102(2): 456-474, 2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-31616913

RESUMO

Survival and growth of the bovine conceptus (embryo and associated extraembryonic membranes) are dependent on endometrial secretions or histotroph found in the uterine lumen. Previously, serial embryo transfer was used to classify heifers as high fertile (HF), subfertile (SF), or infertile (IF). Here, we investigated specific histotroph components [glucose, prostaglandins (PGs), and lipids] in the uterine lumen of day 17 pregnant and open fertility-classified heifers. Concentrations of glucose in the uterine lumen were increased by pregnancy but did not differ among fertility-classified heifers. Differences in expression of genes encoding glucose transporters and involved with glycolysis and gluconeogenesis were observed between conceptuses collected from HF and SF heifers. In the uterine lumen, PGE2 and PGF2α were increased by pregnancy, and HF heifers had higher concentrations of PGE2, PGF2α, and 6-keto-PFG1α than SF heifers. Differences were found in expression of genes regulating PG signaling, arachidonic acid metabolism, and peroxisome proliferator-activated receptor signaling among conceptuses and endometrium from fertility-classified heifers. Lipidomics was conducted exclusively in samples from HF heifers, and phosphatidylcholine was the main lipid class that increased in the uterine lumen by pregnancy. Expression of several lipid metabolism genes differed between HF and SF conceptuses, and a number of fatty acids were differentially abundant in the uterine lumen of pregnant HF and SF heifers. These results support the ideas that uterine luminal histotroph impacts conceptus survival and programs its development and is a facet of dysregulated conceptus-endometrial interactions that result in loss of the conceptus in SF cattle during the implantation period of pregnancy establishment.


Assuntos
Glucose/metabolismo , Lipídeos/análise , Prenhez/metabolismo , Prostaglandinas/metabolismo , Útero/metabolismo , Animais , Bovinos , Implantação do Embrião/fisiologia , Transferência Embrionária/veterinária , Desenvolvimento Embrionário/fisiologia , Endométrio/metabolismo , Feminino , Expressão Gênica , Gluconeogênese/genética , Glicólise/genética , Gravidez , Proteínas de Transporte de Sódio-Glucose/genética , Proteínas de Transporte de Sódio-Glucose/metabolismo
19.
Theriogenology ; 142: 384-389, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31708196

RESUMO

The objective was to determine the effects of pregnancy status on oxylipin profiles and eicosanoid metabolizing enzymes and in corpora lutea (CL) or endometrial (caruncle; CAR and intercaruncle; IC) tissues. Angus crossed cattle were synchronized with the CO-Synch protocol and artificially inseminated (AI). Sixteen days after AI, cattle were euthanized, and reproductive tracts collected from 6 non-pregnant and 6 pregnant cows. Oxylipin profiles and concentrations of progesterone (P4) were obtained from CL tissues. The activity of cytochrome P450 1A (CYP1A) and UDP-glucuronosyltransferase (UGT) enzymes were determined using specific luminogenic substrates. Data were analyzed using the MIXED procedure of SAS, and the model included pregnancy status. Corpora lutea of pregnant cattle contained greater (P < 0.05) concentrations of 9,10-DiHODE, 15,16-DiHODE, and 9,10-DiHOME. These oxylipins have been observed to increase cellular proliferation and vasodilation. Activity of CYP1A in the CL and UGT in CAR and IC was not different (P > 0.05) between pregnant and non-pregnant cattle. In the CL, activity of UGT was decreased (P < 0.05) in pregnant vs. non-pregnant cattle. The decrease in CL UGT activity during pregnancy indicates alterations in local hormone metabolism, while no differences in CL weight nor amount of P4 in CL were different between pregnant and non-pregnant cattle. Moreover, the increase in specific concentrations of oxylipins in the CL may indicate a novel pathway of steroid and eicosanoid metabolism during maternal recognition of pregnancy.


Assuntos
Corpo Lúteo/química , Oxilipinas/análise , Prenhez , Animais , Bovinos , Corpo Lúteo/metabolismo , Endométrio/metabolismo , Feminino , Inseminação Artificial/veterinária , Relações Materno-Fetais , Oxilipinas/metabolismo , Gravidez , Prenhez/metabolismo
20.
Anat Sci Int ; 95(2): 277-285, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31865549

RESUMO

Several investigations have shown that pregnancy and lactation are able to induce elongation of long bone by altering epiphyseal cartilage function in a prolactin-dependent manner. Since the transcription factor Sox9 is of utmost importance for chondrocyte proliferation and differentiation and since bromocriptine, a dopaminergic D2 agonist widely used to suppress milk production, is known to disrupt the production and release of prolactin, we herein aimed to investigate whether pregnancy and lactation as well as bromocriptine could alter the expression of Sox9. Our immunohistochemical analysis showed that the Sox9 expression levels were markedly upregulated in the tibial proliferative zone of day 21 pregnant rats. In day 8 (early) and day 14 (mid) lactating rats, the Sox9 expression was enhanced only in the proliferative zone, but not in the resting and hypertrophic zones. There was no change in Sox9 expression in day 21 (late) lactating rats. Postweaning rats manifested a decreased Sox9 expression in the hypertrophic zone. Bromocriptine had no effect on Sox9 expression in the proliferative zone of day 21 pregnant rats; however, it completely prevented the Sox9 upregulation in those of early and mid-lactating rats. A differential response was observed in the proliferative and hypertrophic zones of late lactating rats, in which bromocriptine enhanced Sox9 expression. Further investigation of cartilaginous matrix revealed no change in proteoglycans accumulation in lactating rats. In conclusion, the upregulated Sox9 expression predominantly occurred in the proliferative zone during late pregnancy and early and mid-lactation, while the bromocriptine effects depended on the periods and epiphyseal zones.


Assuntos
Bromocriptina/farmacologia , Expressão Gênica/efeitos dos fármacos , Lâmina de Crescimento/metabolismo , Lactação/metabolismo , Prenhez/metabolismo , Proteoglicanas/metabolismo , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Animais , Feminino , Lactação/genética , Gravidez , Prenhez/genética , Ratos , Regulação para Cima/efeitos dos fármacos
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