RESUMO
Changes in endometrial cell morphology and function are absolutely necessary for successful embryo implantation. In this study, miR-26a was widely expressed in dairy goats, and was found to be regulated by ß-estradiol (E2) and progesterone (P4) in endometrial epithelium cells (EECs) as well as stromal cells (ESCs). Furthermore, miR-26a played a role in the regulation of cells proliferation and apoptosis by directly regulating PTEN and indirectly regulating the PI3K/AKT pathway in EECs but not in ESCs of dairy goats in vitro. In addition, miR-26a regulated the expression of osteopontin (OPN), vascular endothelial growth factor (VEGF), Cyclooxygenase-2 (COX-2), and prolactin (PRL) in endometrial cells. Therefore, we could get a conclusion that miR-26a had very complex and diverse functions in the endometrial cells during the development of endometrial receptivity in dairy goats. This study provided an efficient platform for studying the regulatory effect of miR-26a on endometrial cells during the development of endometrial receptivity in dairy goats.
Assuntos
Apoptose , Proliferação de Células , Endométrio/enzimologia , Células Epiteliais/enzimologia , MicroRNAs/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células Estromais/enzimologia , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Indústria de Laticínios , Endométrio/efeitos dos fármacos , Endométrio/embriologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/ultraestrutura , Estradiol/farmacologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Idade Gestacional , Cabras , MicroRNAs/genética , Osteopontina/genética , Osteopontina/metabolismo , Gravidez , Progesterona/farmacologia , Prolactina/genética , Prolactina/metabolismo , Transdução de Sinais , Células Estromais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Increasing evidence has shown that miRNAs play important roles in endometrium development during the menstrual cycle in humans and many other animals. Our previous data indicated that miR-182 levels increase 15.55-fold and pleiotrophin (PTN) levels decrease 20.97-fold in the receptive endometrium (RE, D15) compared with the pre-receptive endometrium (PE, D5) in dairy goats. The present study shows that miR-182 is widely expressed in different tissues of dairy goats and that its expression levels are regulated by E2 and P4 in endometrial epithelium cells (EECs). We confirmed that PTN is a target of miR-182 and that miR-182 regulates the protein levels of AKT, Bcl-2, FAS, MAPK, Caspase-3 and SP1 in EECs. Furthermore, miR-182 up-regulates or maintains the expression levels of osteopontin (OPN), cyclooxygenase-2 (COX-2) and prolactin receptor (PRLR) in EECs, suggesting that miR-182 is an important regulatory factor in the construction of endometrial receptivity in dairy goats. In conclusion, miR-182 participates in the development of endometrial receptivity by down-regulating PTN and affecting the expression of select apoptosis-related genes and increasing or maintaining the expression levels of OPN, COX-2 and PRLR in the EECs of dairy goats.
Assuntos
Proteínas de Transporte/genética , Citocinas/genética , Endométrio/metabolismo , Cabras/metabolismo , MicroRNAs/fisiologia , Regiões 3' não Traduzidas , Animais , Apoptose , Sequência de Bases , Sítios de Ligação , Proteínas de Transporte/metabolismo , Proliferação de Células , Células Cultivadas , Citocinas/metabolismo , Regulação para Baixo , Endométrio/citologia , Células Endoteliais/fisiologia , Estradiol/fisiologia , Ciclo Estral , Feminino , Expressão Gênica , Cabras/genética , Progesterona/fisiologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNARESUMO
Under natural conditions, some embryos cannot implant successfully because of the dysfunction of receptive endometrium (RE). Thus, it is imperative for us to study the molecular mechanisms involved in the formation of the RE from pre-receptive endometrium (PE). In this study, the endometrium from gestational day 5 (D5, PE) and gestational day 15 (D15, RE) dairy goats were selected to systematically analyze the transcriptome using strand-specific Ribo-Zero RNA-Seq, >120 million high-quality paired-end reads were generated and 47,616 transcripts were identified in the endometrium of dairy goats. A total of 810 mRNAs were differentially expressed genes (DEGs) between the RE and PE meeting the criteria of P-values<0.05. Bioinformatics analysis of the DEGs revealed that a number of biological processes and pathways were potentially involved in the establishment of the RE, notably energy metabolism and amino acid metabolism. Furthermore, we speculated that CXCL14, IGFBP3, and LGALS15 potentially participated in the development of endometrium. What's more, putative SNPs, InDels and AS events were identified and analyzed in the endometrium. In a word, this resulting view of the transcriptome greatly enhances the comprehensive transcript catalog and uncovers the global trends in gene expression during the formation of receptive endometrium in dairy goats.
Assuntos
Implantação do Embrião/genética , Endométrio/metabolismo , Cabras/embriologia , Cabras/genética , Transcriptoma , Animais , Quimiocinas CXC/genética , Feminino , Galectinas/genética , Perfilação da Expressão Gênica , Idade Gestacional , Mutação INDEL , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/metabolismo , Análise de Sequência de RNARESUMO
We used 240 kid Boer goats that were divided into six groups. The control group was fed a basal diet containing 0.05 mg of selenium (Se)/kg dry matter (DM). Trial groups received the basal diet supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5 mg Se/kg DM (using a commercial selenomethionine product). Trial groups showed an improvement in growth performance (P < 0.05) despite no change in average daily feed intakes (ADFIs) (P > 0.05) compared to the control group A, quadratic model showed a correlation between glutathione peroxidase activity level in whole blood and dietary Se concentration (R(2) = 0.883, P < 0.04). The best linear model showed that increasing concentrations of Se in the blood (R(2) = 0.968, P < 0.001) and muscle (R(2) = 0.942, P < 0.001) corresponded to increasing Se concentrations in feed. Accumulation of Se in different tissues and organs corresponded to increasing Se concentrations in the diet as well as to the total time goats spent feeding on supplemented diet. Kidney and muscle tissues showed the highest and lowest accumulation of Se, respectively. Thus, Se in goat meat can be increased by adding between 0.1 and 0.5 mg/kg of selenomethionine to the diet of goats.
Assuntos
Suplementos Nutricionais , Glutationa Peroxidase/sangue , Cabras/sangue , Selênio/sangue , Selenometionina/farmacologia , AnimaisRESUMO
BACKGROUND: MicroRNAs (miRNAs) are short, highly conserved small noncoding RNAs that had fundamental roles in post-transcriptional gene expression, and they are crucial for proper control of biological processes and known to participate in embryo implantation. However, miRNA expression profiles in the pre-receptive and receptive phases of the goat endometrium during embryo implantation are unknown. RESULTS: A total of 1,069 and 847 miRNAs were expressed in receptive (R) and pre-receptive (P) goat endometrium, and 632 miRNAs were co-expressed in both phases. We identified 545 (50.98%) known miRNAs in the R library and 522 (61.63%) in the P library. There were 110 up-expressed miRNAs and 33 down-expressed miRNAs in receptive endometrium compared with the pre-receptive endometrium meeting the criteria of P-values< 0.05. Moreover, GO and KEGG analysis of the target genes of the differentially expressed miRNAs revealed some candidate miRNAs, genes and pathways that may involve in the formation of the receptive endometrium. Based on stem-loop RT-qPCR, 15 miRNAs were detected and the results suggested that the majority of the miRNA expression data measured by Solexa deep sequencing could represent actual miRNA expression levels. CONCLUSIONS: Our data revealed the first miRNA profile related to the biology of the goat receptive endometrium during embryo implantation, and the results suggested that a subset of miRNAs might play important roles in the formation of endometrial receptivity. Thus, elucidating the physiological roles of endometrial miRNAs will help us better understand the genetic control of embryo implantation in goats.
Assuntos
Implantação do Embrião , Endométrio/metabolismo , Cabras/metabolismo , MicroRNAs/metabolismo , Animais , Sequência de Bases , Feminino , Cabras/genética , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs/química , Conformação de Ácido Nucleico , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA , TranscriptomaRESUMO
Endometrium receptivity is essential for successful embryo implantation in mammals. However, the lack of genetic information remains an obstacle to understanding the mechanisms underlying the development of a receptive endometrium from the pre-receptive phase in dairy goats. In this study, more than 4 billion high-quality reads were generated and de novo assembled into 102,441 unigenes; these unigenes were annotated using published databases. A total of 3,255 unigenes that were differentially expressed (DEGs) between the PE and RE were discovered in this study (P-values < 0.05). In addition, 76,729-77,102 putative SNPs and 12,837 SSRs were discovered in this study. Bioinformatics analysis of the DEGs revealed a number of biological processes and pathways that are potentially involved in the establishment of the RE, notably including the GO terms proteolysis, apoptosis, and cell adhesion and the KEGG pathways Cell cycle and extracellular matrix (ECM)-receptor interaction. We speculated that ADCY8, VCAN, SPOCK1, THBS1, and THBS2 may play important roles in the development of endometrial receptivity. The de novo assembly provided a good starting point and will serve as a valuable resource for further investigations into endometrium receptivity in dairy goats and future studies on the genomes of goats and other related mammals.
Assuntos
Endométrio/metabolismo , Perfilação da Expressão Gênica , Cabras/genética , Transcriptoma , Animais , Análise por Conglomerados , Biologia Computacional/métodos , Feminino , Regulação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Repetições de Microssatélites , Anotação de Sequência Molecular , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Previous studies revealed that Se was an important regulatory factor for glutathione peroxidase (GPx) genes. However, the relationship between Se concentrations and mRNA expression levels of GPxs were unclear in goats, especially the goats living in natural Se-enriched area. Thus, the aim of this study was to determine the Se concentrations and the mRNA expression levels of GPx-1, GPx-2, GPx-3, and GPx-4 in goats from Ziyang County (ZY-H and ZY-L goats) and Baoji City (BJ-P goats), which were Se-rich region and Se-poor region in China, respectively. Atomic fluorescence spectrometry was used as an essential method to determine the Se concentrations in heart, liver, spleen, lung, kidney, longissimus, biceps femoris, and serum, and the gene expressions were quantified in mRNA samples extracted from the above tissues by real-time quantitative reverse transcription-polymerase chain reaction. We found that the Se concentrations in ZY-H and ZY-L goats were higher than that in BJ-P goats significantly (P < 0.05), and the pertinence relations of Se levels between serum and heart, liver, spleen, and kidney were significant (P < 0.05). The mRNA levels of GPx-1 in ZY-H and ZY-L goats were higher than that in BJ-P goats very significantly (P < 0.01) except for longissimus (P < 0.05). Our results indicated a significant trend for GPx-2 in the direction of increasing mRNA levels with increasing Se concentrations in goats but had no statistical significance (P > 0.05) in our experimental conditions. As to GPx-3, its mRNA expression in spleen, lung, and kidney (P < 0.05) were upregulated and were consensual to high Se contents in ZY-H goats, but no significant effects were observed in heart, liver, longissimus, and biceps femoris among our three groups (P > 0.05). The mRNA levels of GPx-4 in heart, liver, lung, and kidney of ZY-H and ZY-L goats were higher than that of BJ-P goats (P < 0.05), and the difference was very significant in lung especially (P < 0.01), but no change in spleen, longissimus, and biceps femoris (P > 0.05). In summary, these data suggested that the goats living in Ziyang County were rich in Se, and the deposition Se played important roles in the mRNA expression of GPx-1, GPx-3, and GPx-4 in certain tissues of goats differentially.