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1.
Theriogenology ; 142: 158-168, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31593883

RESUMO

SIRT6, a member of the sirtuin family, is a NAD + dependent protein deacetylase and has been implicated in transcriptional regulation of somatic cells and post-transcriptional regulation of oocyte meiosis. However, the function of cumulus cell-derived and maternal SIRT6 in meiotic maturation of porcine oocytes is not yet known. Here, we report that SIRT6 mRNA and protein exists in the oocyte and its surrounding cumulus cells during meiotic maturation. Functional studies using a specific inhibitor in cumulus-enclosed oocytes revealed important roles for SIRT6 in germinal vesicle breakdown (GVBD) and cumulus expansion. Moreover, inhibitor treatment led to a significant reduction in the rate of first polar body (PB1) extrusion and early development of parthenogenetically activated embryos. In contrast, SIRT6 inhibition in cumulus-free oocytes only resulted in a significant reduction in the rate of PB1 extrusion. Furthermore, SIRT6 dysfunction regardless of the origin in both cumulus cells and oocytes severely impaired spindle organization and chromosome alignment at the metaphase stage. Molecularly, SIRT6 inhibition in cumulus cells significantly reduced expression of genes associated with cumulus expansion and gap junctional communication and even expression levels of active phosphorylated CDK1 in oocytes. Importantly, adenylate cyclase inhibition could partially rescue GVBD and PB1 extrusion in SIRT6-inhibited cumulus-enclosed oocytes. Taken together, these results demonstrate that cumulus cell-expressed and maternal SIRT6 differentially regulates porcine oocyte meiotic maturation.

2.
Theriogenology ; 142: 421-432, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31711705

RESUMO

AMH (Anti-Müllerian Hormone) is involved in the regulation of follicle growth initiation and inhibits FSH-induced aromatase expression and estrogen production in granulosa cells. However, the function of AMH in steroidogenesis by theca cells remains unclear. The aim of this study is to investigate the role of AMH as a regulator of the basal and stimulated steroid production by pig granulosa cells (pGCs) and theca cells (pTCs). PGCs and pTCs were incubated with hormones AMH, LH (luteinizing hormone), FSH (follicle stimulating hormone), individually or in combination. The expression of CYP19A1, HSD3B1, CYP11A1, LHCGR, and CYP17A1 mRNA were evaluated by quantitative reverse transcriptase PCR. In pGCs, 10 ng/mL AMH significantly decreased the FSH-stimulated effect on FSHR and CYP19A1 expression and estradiol production. In pTCs, LH treatment significantly increased the expression of HSD3B1, CYP11A1, LHCGR, and androstenedione or progesterone production (P < 0.05). Additionally, 10 ng/mL AMH also significantly decreased the LH-stimulated effects on the expression of HSD3B1, CYP11A1, CYP17A1, LHCGR and androstenedione production. Transfection with siAMHR2-I abolished the suppressive effects of AMH on LH-induced HSD3B1 expression and androstenedione production. Taken together, these results demonstrate that AMH is involved in FSH induced estradiol production in pGCs and LH induced androstenedione production in pTCs by regulating the steroidogenesis pathway.

3.
Cells ; 8(12)2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31835702

RESUMO

The establishment of a functional trophectoderm (TE) epithelium is an essential prerequisite for blastocyst formation and placentation. Transcription coactivator yes-associated protein (YAP), a downstream effector of the hippo signaling pathway, is required for specification of both the TE and epiblast lineages in mice. However, the biological role of YAP in porcine blastocyst development is not known. Here, we report that maternally derived YAP protein is localized to both the cytoplasm and nuclei prior to the morula stage and is then predominantly localized to the TE nuclei in blastocysts. Functionally, maternal YAP knockdown severely impeded blastocyst formation and perturbed the allocation of the first two lineages. The treatment of embryos with verteporfin, a pharmacological inhibitor of YAP, faithfully recapitulated the phenotype observed in YAP deleted embryos. Mechanistically, we found that maternal YAP regulates multiple genes which are important for lineage commitment, tight junction assembly, and fluid accumulation. Consistent with the effects on tight junction gene expression, a permeability assay revealed that paracellular sealing was defective in the trophectoderm epithelium. Lastly, YAP knockdown in a single blastomere at the 2-cell stage revealed that the cellular progeny of the YAP+ blastomere were sufficient to sustain blastocyst formation via direct complementation of the defective trophectoderm epithelium. In summary, these findings demonstrate that maternal YAP facilitates porcine blastocyst development through transcriptional regulation of key genes that are essential for lineage commitment, tight junction assembly, and fluid accumulation.

4.
Front Genet ; 10: 1040, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31708972

RESUMO

Pre-implantation development is a dynamic, complex and precisely regulated process that is critical for mammalian development. There is currently no description of the role of the long noncoding RNAs (lncRNAs) during the pre-implantation stages in the goat. The in vivo transcriptomes of oocytes (n = 3) and pre-implantation stages (n=19) at seven developmental stages in the goat were analyzed by RNA sequencing (RNA-Seq). The major zygotic gene activation (ZGA) event was found to occur between the 8- and 16-cell stages in the pre-implantation stages. We identified 5,160 differentially expressed lncRNAs (DELs) in developmental stage comparisons and functional analyses of the major and minor ZGAs. Fourteen lncRNA modules were found corresponding to specific pre-implantation developmental stages by weighted gene co-expression network analysis (WGCNA). A comprehensive analysis of the lncRNAs at each developmental transition of high correlation modules was done. We also identified lncRNA-mRNA networks and hub-lncRNAs for the high correlation modules at each stage. The extensive association of lncRNA target genes with other embryonic genes suggests an important regulatory role for lncRNAs in embryonic development. These data will facilitate further exploration of the role of lncRNAs in the developmental transformation in the pre-implantation stage.

5.
Expert Opin Ther Targets ; 23(11): 967-986, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31711309

RESUMO

Introduction: Inflammation resolution induced by specialized pro-resolving lipid mediators (SPMs) is a new concept. The application of SPMs is a promising therapeutic strategy that can potentially supersede anti-inflammatory drugs. Most CNS diseases are associated with hyperreactive inflammatory damage. CNS inflammation causes irreversible neuronal loss and permanent functional impairments. Given the high mortality and morbidity rates, the investigation of therapeutic strategies to ameliorate inflammatory damage is necessary.Areas covered: In this review, we explore inflammation resolution in CNS disorders. We discuss the underlying mechanisms and dynamic changes of SPMs and their precursors in neurological diseases and examine how this can potentially be incorporated into the clinic. References were selected from PubMed; most were published between 2010 and 2019.Expert opinion: Inflammation resolution is a natural process that emerges after acute or chronic inflammation. The evidence that SPMs can effectively ameliorate hyperreactive inflammation, shorten resolution time and accelerate tissue regeneration in CNS disorders. Adjuvants and nanotechnology offer opportunities for SPM drug design; however, more preclinical studies are necessary to investigate basic, critical issues such as safety.

6.
Water Environ Res ; 2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31650659

RESUMO

A p-type TiO2 with Ti vacancies (D-TiO2 ) was synthesized by a facile solvothermal treatment, and Ag/TiO2 with different Ag loading amount was prepared through a photo-reduction deposition method. The samples were characterized through scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The adsorption and photocatalytic characteristics of tetrabromobisphenol A (TBBPA) on D-TiO2 and Ag/TiO2 were investigated. The adsorption of TBBPA on Ag/TiO2 was significantly enhanced and was five times greater than that of pure TiO2 . The increase in pH significantly inhibited the adsorption of TBBPA. The 2%-Ag/TiO2 nearly completely degraded TBBPA in 10 min under UV-Vis light (λ > 360 nm), and the apparent reaction rate constant (kapp ) reached 0.63 min-1 . The significantly enhanced UV-Vis light catalytic properties of the Ag/TiO2 in comparison with that of TiO2 were attributed to the increased adsorption capacity and electron transfer ability of the Ag/TiO2 . Free radical trap experiments results showed that holes and superoxide radicals play a major role in the catalytic degradation of TBBPA by Ag/TiO2 . Moreover, the Ag/TiO2 catalyst exhibits high stability during TBBPA degradation even after three cycles. PRACTITIONER POINTS: Ti-defected TiO2 and Ag/TiO2 were synthesized using a solvothermal and photo-reduction deposition, respectively. Ag/TiO2 exhibited outstanding adsorption and photocatalytic activity for TBBPA removal under UV-Vis light. Holes and superoxide radicals play a major role in the photocatalytic degradation of TBBPA.

7.
Aging (Albany NY) ; 11(18): 8015-8034, 2019 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-31562810

RESUMO

Thousands of circular RNAs (circRNAs) have been recently discovered in cumulus cells and oocytes from several species. However, the expression and function of circRNA during porcine oocyte meiotic maturation have been never examined. Here, we separately identified 7,067 and 637 circRNAs in both cumulus cells and oocytes via deep sequencing and bioinformatic analysis. Further analysis revealed that a faction of circRNAs is differentially expressed (DE) in a developmental stage-specific manner. The host genes of DE circRNAs are markedly enriched to multiple signaling pathways associated with cumulus cell function and oocyte maturation. Additionally, most DE circRNAs harbor several miRNA targets, suggesting that these DE circRNAs potentially act as miRNA sponge. Importantly, we found that maternal circARMC4 knockdown by siRNA microinjection caused a severely impaired chromosome alignment, and significantly inhibited first polar body extrusion and early embryo development. Taken together, these results demonstrate for the first time that circRNAs are abundantly and dynamically expressed in a developmental stage-specific manner in cumulus cells and oocytes, and maternally expressed circARMC4 is essential for porcine oocyte meiotic maturation and early embryo development.

8.
Sci Rep ; 9(1): 13186, 2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31515498

RESUMO

Swine enteric diseases have caused significant economic loss and have been considered as the major threat to the global swine industry. Several coronaviruses, including transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV), have been identified as the causative agents of these diseases. Effective measures to control these diseases are lacking. The major host cells of transmissible gastroenteritis virus and porcine epidemic diarrhea virus have thought to be epithelial cells on small intestine villi. Aminopeptidase-N (APN) has been described as the putative receptor for entry of transmissible gastroenteritis virus and porcine epidemic diarrhea virus into cells in vitro. Recently, Whitworth et al. have reported that APN knockout pigs are resistant to TGEV but not PEDV after weaning. However, it remains unclear if APN-null neonatal pigs are protected from TGEV. Here we report the generation of APN-null pigs by using CRISPR/Cas9 technology followed by somatic cell nuclear transfer. APN-null pigs are produced with normal pregnancy rate and viability, indicating lack of APN is not embryonic lethal. After viral challenge, APN-null neonatal piglets are resistant to highly virulent transmissible gastroenteritis virus. Histopathological analyses indicate APN-null pigs exhibit normal small intestine villi, while wildtype pigs show typical lesions in small intestines. Immunochemistry analyses confirm that no transmissible gastroenteritis virus antigen is detected in target tissues in APN-null piglets. However, upon porcine epidemic diarrhea virus challenge, APN-null pigs are still susceptible with 100% mortality. Collectively, this report provides a viable tool for producing animals with enhanced resistance to TGEV and clarifies that APN is dispensable for the PEDV infection in pigs.

9.
R Soc Open Sci ; 6(7): 190351, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31417737

RESUMO

In this study, natural manganese oxides (MnO x ), an environmental material with high redox potential, were used as a promising low-cost oxidant to degrade the widely used dyestuff methylene blue (MB) in aqueous solution. Although the surface area of MnO x was only 7.17 m2 g-1, it performed well in the degradation of MB with a removal percentage of 85.6% at pH 4. It was found that MB was chemically degraded in a low-pH reaction system and the degradation efficiency correlated negatively with the pH value (4-8) and initial concentration of MB (10-50 mg l-1), but positively with the dosage of MnO x (1-5 g l-1). The degradation of MB fitted well with the second-order kinetics. Mathematical models were also built for the correlation of the kinetic constants with the pH value, the initial concentration of MB and the dosage of MnO x . Furthermore, several transformation products of MB were identified with HPLC-MS, which was linked with the bond energy theory to reveal that the degradation was initiated with demethylation.

10.
Int J Mol Sci ; 20(16)2019 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-31416143

RESUMO

A series of complex processes regulate muscle development, and lncRNAs play essential roles in the regulation of skeletal myogenesis. Using RNA sequencing, we profiled the lncRNA expression during goat (Capra hircus) skeletal muscle development, which included seven stages across fetal 45 (F45), 65 (F65), 90 (F90), 120 (F120), 135 (F135) days, born for 24 h (B1) and 90 (B90) days. A total of 15,079 lncRNAs were identified in the seven stages, and they were less conservative with other species (human, cow, and mouse). Among them, 547 were differentially expressed, and they divided the seven stages into three functional transition periods. Following weighted gene co-expression network analysis (WGCNA), five lncRNA modules specific for developmental stages were defined as three types: 'Early modules', 'late modules', and 'individual-stage-specific modules'. The enrichment content showed that 'early modules' were related to muscle structure formation, 'late modules' participated in the 'p53 signaling pathway' and other pathways, the F90-highly related module was involved in the 'MAPK signaling pathway', and other pathways. Furthermore, we identified hub-lncRNA in three types of modules, and LNC_011371, LNC_ 007561, and LNC_001728 may play important roles in goat skeletal muscle. These data will facilitate further exploration of skeletal muscle lncRNA functions at different developmental stages in goats.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Cabras/genética , Desenvolvimento Muscular/genética , Músculo Esquelético/metabolismo , RNA Longo não Codificante/genética , Animais , Biomarcadores , Diferenciação Celular/genética , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Sequenciamento de Nucleotídeos em Larga Escala , RNA Mensageiro/genética , Reprodutibilidade dos Testes , Transdução de Sinais , Transcriptoma
11.
Anim Biotechnol ; : 1-8, 2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-31253059

RESUMO

Pleomorphic adenoma gene 1 (PLAG1) encodes a developmentally regulated zinc finger protein, locating in growth-related QTNs. The mRNA expression of this gene was investigated in different tissues and from two different developmental periods, whilst to explore the functions of PLAG1 in growth traits of cattle. The results showed that PLAG1 was expressed in all examined tissues. However, PLAG1 expression levels in all examined tissues were significantly different between the 5-month fetus and 36-month adult cattle. Our juvenile results indicated PLAG1 is primarily expressed in embryonic tissues of Chinese cattle. Furthermore, two variations were identified. Association analysis revealed that the two variations were associated with growth traits (p < 0.05 or p < 0.01). These new findings provide a comprehensive overview of the critical roles of PLAG1 in growth traits modulation and can be highlighted as candidate molecular markers in cattle breeding.

12.
Glob Chang Biol ; 25(9): 2958-2969, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31152626

RESUMO

Nitrogen (N) enrichment often increases aboveground net primary productivity (ANPP) of the ecosystem, but it is unclear if belowground net primary productivity (BNPP) track responses of ANPP. Moreover, the frequency of N inputs may affect primary productivity but is rarely studied. To assess the response patterns of above- and belowground productivity to rates of N addition under different addition frequencies, we manipulated the rate (0-50 g N m-2  year-1 ) and frequency (twice vs. monthly additions per year) of NH4 NO3 inputs for six consecutive years in a temperate grassland in northern China and measured ANPP and BNPP from 2012 to 2014. In the low range of N addition rates, BNPP showed the greatest negative response and ANPP showed the greatest positive responses with increases in N addition (<10 g N m-2  year-1 ). As N addition increased beyond 10 g N m-2  year-1 , increases in ANPP dampened and decreases in BNPP ceased altogether. The response pattern of net primary productivity (combined above- and belowground; NPP) corresponded more closely to ANPP than to BNPP. The N effects on BNPP and BNPP/NPP (fBNPP ) were not dependent on N addition frequency in the range of N additions typically associated with N deposition. BNPP was more sensitive to N addition frequency than ANPP, especially at low rates of N addition. Our findings provide new insights into how plants regulate carbon allocation to different organs with increasing N rates and changing addition frequencies. These root response patterns, if incorporated into Earth system models, may improve the predictive power of C dynamics in dryland ecosystems in the face of global atmospheric N deposition.


Assuntos
Ecossistema , Nitrogênio , Carbono , China , Plantas
13.
Anim Biotechnol ; : 1-8, 2019 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-31179830

RESUMO

MicroRNA-1 (miR-1) has been shown to play an important role in muscle growth and development, however, it was mainly discovered in model animals. To explore the function and mechanism of miR-1 in goat, we firstly explored the expression profile of miR-1 in goat tissues and cells. Furthermore, the target gene of miR-1 was predicted, and the relationship between miR-1 and one of its target genes, histone deacetylase 4 (HDAC4), was analyzed through double luciferase reporter assay, real-time PCR, and western blot. It was found that the miR-1 is most abundantly expressed in goat heart and skeletal muscle tissue. Meanwhile, the expression of miR-1 showed an increasing tendency from new-born goats to the 7-month-old goats, and then its expression decreases as the goats mature further. In addition, the expression levels of miR-1 decreased in goat skeletal muscle satellite cells with the algebraic increasing of cells. At last, the results showed that HDAC4 is a target gene of miR-1 in goat, and miR-1 can inhibit the post-transcriptional expression of HDAC4, but had no significant influence on the mRNA level of HDAC4. It was hypothesized that miR-1 promotes muscle development by inhibiting the post-transcriptional expression of HDAC4 in goat.

14.
BMC Neurol ; 19(1): 133, 2019 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-31215399

RESUMO

BACKGROUND: Heat stroke (HS) is a critical illness that can cause multiple organ dysfunction, including damage to the central nervous system (CNS), which can be life-threatening in severe cases. Brain lesions in patients with HS who present with CNS damage have been rarely reported before, and they usually vary in different cases, hence, patients with such lesions may present a clinical challenge in terms of diagnosis and management. Cerebral venous thrombosis (CVT) is a rare cause of stroke that mostly affects young individuals and children. The pathogenesis of brain damage caused by HS is complex, and CVT may be involved in the pathogenesis of HS with CNS damage. In this manuscript, we have reported a case of a patient with HS having CVT with symmetrical lesions in the bilateral putamen, posterior limb of the internal capsule, external capsule, insular lobe, and subcortical white matter in the brain. CASE PRESENTATION: We encountered a 48-year-old man who presented with HS in the summer season. During admission, he had a high body temperature and was in coma and shock. Then, he developed rhabdomyolysis syndrome, acute kidney and liver damage, electrolyte imbalance, and acid-base balance disorders, and his D-dimer level was elevated. After several days of anti-shock treatment, the patient's level of consciousness improved. However, he experienced a decline in vision. Cerebral magnetic resonance imaging (MRI) showed symmetrical lesions in the bilateral posterior limb of the internal capsule, putamen, external capsule, insula, and subcortical white matter, and cerebral magnetic resonance venography (MRV) showed the development of CVT. Therefore, anti-coagulation treatment was provided. After timely clinical intervention, the symptoms of the patient gradually improved. CONCLUSIONS: This case showed that HS can cause CVT. Therefore, cerebral MRI findings in HS must be assessed; in addition, early MRV can help in the diagnosis of the disease, which can effectively improve prognosis.


Assuntos
Golpe de Calor/complicações , Golpe de Calor/diagnóstico por imagem , Golpe de Calor/patologia , Trombose Intracraniana/diagnóstico por imagem , Trombose Intracraniana/etiologia , Trombose Intracraniana/patologia , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Humanos , Imagem por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Flebografia/métodos
15.
Theriogenology ; 133: 149-158, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31100559

RESUMO

Melatonin has been reported to improve the survival rate of mouse and goat preantral follicles cultured in vitro. However, the role of melatonin in the development of oocyte-granulosa cell complexes (OGCs) isolated from preantral follicles remains unclear. Cumulus-oocyte complexes were isolated from OGCs cultured in vitro for 18.5 days and were then maturated in vitro for 42 h. The matured oocytes were parthenogenetically activated and were further cultured up to the blastocyst stage. We found that the developmental capacity of oocytes from in vitro cultured OGCs was significantly inferior to that from in vivo grown counterparts. Additionally, a 10-5 M dose of melatonin added to the medium during in vitro culture of OGCs did not improve oocyte meiotic maturation but enhanced blastocyst rate of parthenogenetically activated embryos. Besides, these beneficial effects could be reversed by luzindole treatment, a melatonin membrane receptor antagonist. mRNA sequencing analysis further revealed that melatonin caused differential expression of 76 genes of which 75 were upregulated and 1 was downregulated in OGCs. Twelve of the 76 genes were identified as potential regulators of metabolic pathways by functional analysis. Taken together, these results indicate that melatonin improves developmental competence of porcine oocyte-granulosa cell complexes.


Assuntos
Melatonina/farmacologia , Oócitos/efeitos dos fármacos , Suínos/fisiologia , Animais , Técnicas de Cultura de Células/veterinária , Meios de Cultura , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Oócitos/crescimento & desenvolvimento , Folículo Ovariano/citologia
16.
Reproduction ; 157(6): 501-510, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30870811

RESUMO

HASPIN kinase-catalyzed phosphorylation of histone H3 on threonine 3 (H3T3p) directs the activity and localization of chromosomal passenger complex (CPC) and spindle assembly checkpoint (SAC) to regulate chromosome condensation and segregation in both mitosis and meiosis. However, the function of HASPIN kinase in the meiotic maturation of porcine oocytes is not yet known. Here, we found that HASPIN mRNA is constantly expressed in porcine oocyte maturation and subsequent early embryo development. H3T3p is highly enriched on chromosomes at germinal vesicle breakdown (GVBD) stage and thereafter maintains a low level in progression through metaphase I (MI) to metaphase II (MII). Correspondingly, H3T3p was completely abolished in oocytes treated with an inhibitor of HASPIN kinase. Functionally, inhibition of HASPIN activity led to a significant reduction in the rate of oocyte meiotic maturation and the limited cumulus expansion. Additionally, HASPIN inhibition caused both spindle disorganization and chromosome misalignment in oocytes at MI and MII stage. Importantly, HASPIN inhibition severely prevented deacetylation of several highly conserved lysine (K) residues of histone H3 and H4 including H3K9, H3K14, H4K5, H4K8, H4K12 and H4K16 on the metaphase chromosomes during oocyte meiotic maturation. Taken together, these results demonstrate that HASPIN kinase regulates porcine oocyte meiotic maturation via modulating histone deacetylation.

17.
Cryobiology ; 87: 91-98, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30707961

RESUMO

Accurate reprogramming of DNA methylation occurring in preimplantation embryos is critical for normal development of both fetus and placenta. Environmental stresses imposed on oocytes usually cause the abnormal DNA methylation reprogramming of early embryos. However, whether oocyte vitrification alters the reprogramming of DNA methylation (5 mC) and its derivatives in mouse preimplantation embryo development remains largely unknown. Here, we found that the rate of cleavage and blastocyst formation of embryos produced by IVF of vitrified matured oocytes was significantly lower than that in control counterparts, but the quality of blastocysts was not impaired by oocyte vitrification. Additionally, although vitrification neither altered the dynamic changes of 5-hydroxymethylcytosine (5hmC) and 5-formylcytosine (5 fC) before 4-cell stage nor affected the levels of 5 mC and 5-carboxylcytosine (5caC) throughout the preimplantation development, vitrification significantly reduced the levels of 5hmC and 5 fC from 8-cell stage onwards. Correspondingly, vitrification did not alter the expression patterns of Tet3 in preimplantation embryos but apparently reduced the expression levels of Tet1 in 4-cell and 8-cell embryos and increased the expression levels of Tet2 at morula stage. Taken together, these results demonstrate that oocyte vitrification perturbs DNA methylation reprogramming in mouse preimplantation embryo development.


Assuntos
Blastocisto/citologia , Criopreservação/métodos , Metilação de DNA/genética , Oócitos/citologia , Oogênese/fisiologia , Vitrificação , 5-Metilcitosina/análogos & derivados , 5-Metilcitosina/química , Animais , Citosina/análogos & derivados , Citosina/química , Proteínas de Ligação a DNA/biossíntese , Dioxigenases/biossíntese , Desenvolvimento Embrionário , Feminino , Fertilização In Vitro/métodos , Metáfase , Camundongos , Mórula/fisiologia , Gravidez , Proteínas Proto-Oncogênicas/biossíntese
18.
J Cell Physiol ; 234(10): 17767-17774, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30805939

RESUMO

NESFATIN-1 acts as a neuroendocrine hormone to suppress gonadotropin secretion in the female goldfish and to prevent germinal vesicle breakdown of oocytes in the zebrafish. However, the expression and function of NESFATIN-1 in meiotic maturation and development of porcine oocytes remains elusive. Genomic structure of porcine NESFATIN-1 precursor nucleobindin 2 (NUCB2) is first characterized in detail and an evolutionally closer relationship of NESFATIN-1 between pig and rat is shown by phylogenetic analysis of multiple species. Additionally, immunofluorescence analysis revealed that NESFATIN-1 is predominantly expressed and localizes on the membrane of both theca cells and granulosa cells, but not expressed in oocytes. Real-time quantitative polymerase chain reaction showed that the abundance of NESFATIN-1 transcripts in granulosa cells progressively decreases during the developmental transition from small follicles to large follicles. Correspondingly, NESFATIN-1 could significantly enhance both the cleavage and blastocyst rate of parthenogenetically activated oocytes from small follicles (p < 0.05), whereas it did not affect meiotic maturation and development of oocytes from large follicles. Interestingly, we found that NESFATIN-1 significantly improves meiotic maturation of oocytes cultured in chemically defined medium in the absence of pyruvate compared with the control group (p < 0.05), suggesting that the NESFATIN-1 as a substitute for pyruvate exerts beneficial effects on porcine oocyte maturation. In conclusion, these results demonstrate that NESFATIN-1 facilitates both meiotic maturation and development of porcine oocytes.

19.
Environ Sci Pollut Res Int ; 26(7): 7055-7070, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30645747

RESUMO

From January 1 to April 22, 2014, an online analyzer for monitoring aerosols and gases (MARGA) was used to measure and analyze water-soluble ions in inhalable particulate matter with a diameter less than 10 µm (PM10) during winter-spring in Shenyang city, China. The results yielded three main findings. (1) During the entire observation period and in seven pollution episodes, SO42-, NO3-, and NH4+ (SNA) accounted for 84.4-93.1% of the total water-soluble ions (TWSIs). TWSIs accounted for 32% of PM10 mass during the entire observation period, and the contribution of TWSIs in PM10 ranged from 33.4-43.1% in the seven pollution episodes. The contribution of TWSIs components increased during the pollution episodes, but certain differences were observed in different pollution episodes. In terms of ionic equilibrium, the total concentration of negative ions was slightly greater than that of positive ions and the difference was 3.1% of the total ion load on average, indicating that local aerosols are mainly neutral. The water-soluble ions show clear diurnal variation with the high concentration around 09:00 for SO42-, NH4+, and Cl- which is consistent with the high heating grade index. (2) Pollution episodes often occur in Northeast China, especially during the winter period. Due to the low temperature in the winter, the local coal burning for heating is one of the main sources of pollution besides vehicle exhaust and industrial pollution, which is supported by the higher NO3-/SO42- ratio in April than that in January to March. Sometimes, under the prevailing wind directions of W and SSW, the long-distance transport of pollutants from the Beijing-Tianjin-Hebei region and Shandong province superimposed on local pollution leads to the most severe pollution, such as Ep3 and Ep5. (3) SO42- concentration is closely related to ambient water vapor pressure (e*), with increase as e* increased depending on the temperature. NO3- concentration showed a linear relationship of excess NH4+, which suggests homogeneous gas-phase reaction of ammonia and nitric acid is possibly an important pathways of nitrate formation in the haze pollution process in Shenyang City. In addition, our results also suggest the nighttime liquid-phase reaction may cause large increases of nitrate in the haze pollution process.


Assuntos
Poluentes Atmosféricos/análise , Poluição do Ar/estatística & dados numéricos , Monitoramento Ambiental , Material Particulado/análise , Aerossóis , China , Cidades , Carvão Mineral , Gases , Nitratos , Óxidos de Nitrogênio , Tamanho da Partícula , Estações do Ano , Emissões de Veículos , Vento
20.
Anim Sci J ; 90(3): 366-371, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30623527

RESUMO

Porcine viral diarrhea is an acute and highly contagious enteric disease in pigs which causes huge economic losses in pig industry worldwide. Transmissible gastroenteritis virus (TGEV) is main pathogens responsible for piglets viral diarrhea. Knockout the host cellular surface receptor for TGEV may be an effective way to accelerate the breeding of resistant pigs. In this study, we applied site-specific editing pAPN which is effective in swine testis (ST) cells. Site-specific editing of pAPN reduced TGEV proliferation in ST cells by 96%-99% at different time periods post-infection. Next, the site-specific editing of pAPN porcine fetal fibroblasts were produced, and then the cell colonies were used as donor cells to generate the site-specific editing of pAPN pigs. Our research findings will not only offer a more thorough understanding of the pathogenesis of piglet diarrhea and lay the foundation for breeding TGEV-resistant piglets, but also understanding the molecular mechanisms involved in coronaviral infections.


Assuntos
Cruzamento/métodos , Antígenos CD13/genética , Diarreia/prevenção & controle , Diarreia/veterinária , Gastroenterite Suína Transmissível/prevenção & controle , Edição de Genes/métodos , Técnicas de Inativação de Genes/métodos , Receptores Virais/genética , Doenças dos Suínos/prevenção & controle , Suínos , Animais , Diarreia/virologia , Fibroblastos/enzimologia , Gastroenterite Suína Transmissível/virologia , Masculino , Doenças dos Suínos/virologia , Testículo/citologia , Testículo/virologia , Vírus da Gastroenterite Transmissível/patogenicidade , Vírus da Gastroenterite Transmissível/fisiologia , Replicação Viral
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