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1.
Anim Reprod Sci ; 226: 106711, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33540216

RESUMO

The objective of this study was to evaluate placental development during late gestation (day 100) between Chinese Meishan (CM; n = 7) and White crossbred (WC; n = 5) gilts following intrauterine crowding induced by unilaterally hysterectomy-ovariectomy. Gross placental morphology and areolae density as well as histological morphology (i.e., folded bilayer and placental stroma) were analyzed using computer-assisted morphometry for placentas of the smallest and largest fetuses within each litter. There was a breed by fetal size interaction (P < 0.01) for areolae density in which placentas for large CM fetuses had greater areolae density compared to small CM fetuses, but the density of areolae was greater for CM fetuses compared to WC fetuses, irrespective of fetal size. The width of the folded bilayer was greater (P < 0.01) in placentas for WC gilts compared to CM gilts, irrespective of fetal size. Placentas for small fetuses had greater (P < 0.01) folded bilayer width compared to large fetuses, irrespective of breed. The placental stromal width was greater (P < 0.01) in placentas for large fetuses compared to small, irrespective of breed. The difference between stromal width in placentas between divergent-sized littermates, however, was greater (P = 0.05) in WC gilts compared to CM gilts, indicating there was a limited response to intrauterine crowding in CM gilts. These results indicate there is an altered placental development during late gestation in CM compared to WC gilts, thus, there are likely different mechanisms for responding to intrauterine crowding between breeds.


Assuntos
Tamanho da Ninhada de Vivíparos , Prenhez , Suínos/genética , Suínos/fisiologia , Animais , Feminino , Feto , Placenta/fisiologia , Placentação , Gravidez , Prenhez/fisiologia , Útero/fisiologia
2.
Biol Reprod ; 100(1): 71-85, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30107478

RESUMO

Reproductive performance of female pigs that do not receive sufficient colostrum from birth is permanently impaired. Whether lactocrine deficiency, reflected by low serum immunoglobulin immunocrit (iCrit), affects patterns of endometrial gene expression during the periattachment period of early pregnancy is unknown. Here, objectives were to determine effects of low iCrit at birth on the adult endometrial transcriptome on pregnancy day (PxD) 13. On the first day of postnatal life, gilts were assigned to high or low iCrit groups. Adult high (n = 8) and low (n = 7) iCrit gilts were bred (PxD 0), and humanely slaughtered on PxD 13 when tissues and fluids were collected. The endometrial transcriptome was defined for each group using mRNAseq and microRNAseq. Reads were mapped to the Sus scrofa 11.1 genome build. Mature microRNAs were annotated using miRBase 21. Differential expression was defined based on fold change (≥ ±1.5). Lactocrine deficiency did not affect corpora lutea number, uterine horn length, uterine wet weight, conceptus recovery, or uterine luminal fluid estrogen content on PxD 13. However, mRNAseq revealed 1157 differentially expressed endometrial mRNAs in high versus low iCrit gilts. Differentially expressed genes had functions related to solute transport, endometrial receptivity, and immune response. Six differentially expressed endometrial microRNAs included five predicted to target 62 differentially expressed mRNAs, affecting similar biological processes. Thus, lactocrine deficiency on the first day of postnatal life can alter uterine developmental trajectory with lasting effects on endometrial responses to pregnancy as reflected at the level of the transcriptome on PxD 13.


Assuntos
Endométrio/metabolismo , Substâncias de Crescimento/deficiência , Lactação/fisiologia , Prenhez , Suínos , Transcriptoma , Animais , Animais Recém-Nascidos , Colostro/fisiologia , Implantação do Embrião/efeitos dos fármacos , Endométrio/efeitos dos fármacos , Endométrio/patologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Idade Gestacional , Substâncias de Crescimento/farmacologia , Gravidez , Prenhez/genética , Prenhez/metabolismo , Suínos/genética , Suínos/metabolismo , Transcriptoma/efeitos dos fármacos
3.
J Anim Sci ; 96(2): 521-531, 2018 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-29385465

RESUMO

Lactation is a very energy demanding period for sows. The current study provides a better understanding of the biochemical response of first- (n = 246) or second-parity (n = 127) sows during late gestation through lactation and assesses relationships with piglet production and dam reproductive performance. Plasma samples were collected from first- or second-parity dams at late gestation (110 d gestation [d110G]), d 1 post-farrowing (d1PF), and weaning (WN) then analyzed for various stress and protein metabolism compounds, including; creatine, creatine phosphokinase (CPK) activity, creatinine, urea nitrogen, albumin, and lactate. Litter performance was measured as number of piglets nursed and piglet ADG. Post-weaning reproductive performance was assessed by measuring weaning-to-estrus interval (WEI) and subsequent ovulation rate collected at time of harvest. Plasma creatine and CPK activity increased (P < 0.05) between d110G and d1PF. Plasma creatinine decreased (P < 0.05) from d110G through WN in first-parity dams, but remained similar between d110G and d1PF before declining (P < 0.05) at WN in second-parity dams. Plasma urea nitrogen increased (P < 0.05) over the course of the study and was negatively (P < 0.05) associated with piglet ADG at d110G and d1PF and with ovulation rate at d110G (P < 0.05). Similarly, plasma albumin increased (P < 0.05) in first-parity dams over the course of the study, whereas it plateaued (P < 0.05) at d1PF and remained similar (P > 0.10) through WN in second-parity dams. First-parity dams had less (P < 0.05) plasma lactate at d110G than at d1PF or WN. However, second-parity dams had increased (P < 0.05) plasma lactate at d110G and d1PF, then decreased (P < 0.05) levels at WN. Plasma lactate at WN was positively (P < 0.05) associated with WEI in first-parity dams, but negatively (P < 0.05) related to WEI at d1PF in second-parity dams. Plasma lactate levels at all time points were positively (P < 0.05) associated with ovulation rate in second-parity dams. The biochemical profile of these dams differed by parity and merits further investigations into these differences to identify methods to improve physiological response to lactation for improved animal welfare, production, and reproductive performance.


Assuntos
Lactação/fisiologia , Ácido Láctico/sangue , Reprodução , Suínos/fisiologia , Bem-Estar do Animal , Animais , Nitrogênio da Ureia Sanguínea , Estro/fisiologia , Feminino , Paridade , Gravidez , Suínos/sangue , Desmame
4.
Artigo em Inglês | MEDLINE | ID: mdl-28883913

RESUMO

BACKGROUND: During late gestation the placental epithelial interface becomes highly folded, which involves changes in stromal hyaluronan. Hyaluronan is composed of glucoronate and N-acetyl-glucosamine. We hypothesized that supplementing gestating dams with glucosamine during this time would support placental folded-epithelial-bilayer development and increase litter size. In Exp. 1, gilts were unilaterally hysterectomized-ovariectomized (UHO). UHO gilts were mated and then supplemented daily with 10 g glucosamine (n = 16) or glucose (control, n = 17) from d 85 of gestation until slaughter (d 105). At slaughter, the number of live fetuses was recorded and each live fetus and its placenta was weighed. Uterine wall samples adjacent to the largest and smallest fetuses within each litter were processed for histology. In Exp. 2, pregnant sows in a commercial sow farm were supplemented with either 10 g glucosamine or glucose daily from d 85 of gestation to farrowing. Total piglets born and born alive were recorded for each litter. In Exp. 3, the same commercial farm and same protocol were used except that the dose of glucosamine and glucose was doubled to 20 g/d. RESULTS: In Exp. 1, the number of live fetuses tended to be greater in glucosamine-treated UHO gilts (P = 0.098). Placental morphometry indicated that the width of the folded bilayer was greater (P = 0.05) in glucosamine-treated gilts. In Exp. 2, litter size did not differ between glucosamine- and glucose-treated sows. However in Exp. 3, the increased dose of glucosamine resulted in a significant treatment by parity interaction (P ≤ 0.01), in which total piglets born and born alive were greater in glucosamine treated sows of later parity (5 and 6). CONCLUSIONS: These results indicated that glucosamine supplementation increased the width of the folds of the placental bilayer and increased litter size in later parity, intact pregnant commercial sows.

5.
J Anim Sci ; 95(2): 688-696, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28380602

RESUMO

Farrowing induction is a common practice among swine producers to manage timing of farrowing and the labor associated with farrowing. In this experiment, the effect of induction of labor using cloprostenol on Day 114 of gestation ( = 88) was compared to our standard farrowing protocol at USMARC (natural farrowing with induction using cloprostenol on Day 116 if needed, = 82) in gilts and sows up to fourth parity. In a subset of dams ( = 10 each treatment), colostrum was collected within 30 min of birth of the first piglet, and at 4, 8, 12, and 24 h. Colostrum samples were measured for immunoglobulin G (IgG) using the immunoglobulin immunocrit and porcine IgG specific ELISA, and for total protein. Blood samples were collected from each live piglet on d 1 of age and measured using the immunocrit assay, and average immunocrit was calculated for each litter. Total piglets born and born alive, birth and weaning weights, and the stillbirth rate and preweaning mortality rate were also recorded for each litter. Results indicated that induction of farrowing by cloprostenol treatment on d 114 reduced average gestation length by 0.5 to 1 d depending on parity ( < 0.05), and reduced overall colostrum immunocrit, IgG and total protein ( < 0.05). Colostrum immunocrits and IgG concentrations were well correlated ( = 0.89; < 0.01) but IgG was curvilinearly related to total protein. Litter average immunocrits were similar in gilts between treatments, but were reduced in later parity sows induced to farrow using cloprostenol on d 114 of gestation. Total born, born alive, birth and weaning weights, and stillbirth and preweaning mortality rates were unaffected by treatments. In conclusion, induction of farrowing using cloprostenol injection on d 114 reduced colostrum IgG concentrations in dams, but this was reflected in a reduction in litter average immunocrit only in later parity sows. This reduction in litter average immunocrit was not sufficient to influence preweaning mortality, but other effects are possible given the reported influence of colostrum on growth and reproductive traits.


Assuntos
Colostro/química , Imunoglobulina G/sangue , Trabalho de Parto Induzido/veterinária , Paridade , Suínos/fisiologia , Animais , Cloprostenol/farmacologia , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Imunoglobulina G/metabolismo , Luteolíticos/farmacologia , Parto , Gravidez , Suínos/sangue , Suínos/imunologia
6.
Anim Reprod Sci ; 179: 1-9, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28215453

RESUMO

Previous research demonstrated a favorable relationship between the number of follicles detectable in the bovine ovary by ultrasonography and fertility, and bovine females with diminished numbers of antral follicles had smaller reproductive tracts. Therefore, we hypothesized that uterine function would be compromised in beef heifers with diminished numbers of antral follilcles. Angus heifers (n=480) were submitted for ultrasonographic evaluation of antral follicle number at 325 and 355d of age. After the second ultrasonographic examination, 40 pubertal heifers with the greatest average number of antral follicles (30.9±0.7) and 40 pubertal heifers with the lowest average number of antral follicles (14.2±0.7) were synchronized with two i.m. injections of prostaglandin F2α (25mg) administered 11d apart, and heifers were slaughtered on d6 (n=26 heifers/group) or d16 (n=14 heifers/group) of the resultant estrous cycle. The uterus was weighed, flushed for determination of protein content, and representative samples were fixed for determination of endometrial gland morphometry. Heifers in the Low group had fewer surface antral follicles and smaller reproductive tracts than heifers in the High group (P<0.01). Protein content of the uterine flushes was decreased in heifers in the Low group (P<0.01); however, there was no difference in the percent area of the endometrium occupied by endometrial glands. From these results, we conclude that the uterine environment of beef heifers with diminished numbers of antral follicles is less conducive to supporting early embryonic survival.


Assuntos
Bovinos/fisiologia , Folículo Ovariano/diagnóstico por imagem , Útero/fisiologia , Animais , Feminino , Regulação da Expressão Gênica/fisiologia , Folículo Ovariano/fisiologia , Proteínas/genética , Proteínas/metabolismo , Útero/diagnóstico por imagem
7.
Biol Reprod ; 95(5): 108, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27535960

RESUMO

The fetal fluids and uterine flushings of pigs contain higher concentrations of fructose than glucose, but fructose is not detected in maternal blood. Fructose can be synthesized from glucose via enzymes of the polyol pathway, aldose reductase (AKR1B1) and sorbitol dehydrogenase (SORD), transported across cell membranes by solute carriers SLC2A5 and SLC2A8, and converted to fructose-1-phosphate by ketohexokinase (KHK). SLC2A8, SLC2A5, AKR1B1, SORD, and KHK mRNAs and proteins were analyzed using quantitative PCR and immunohistochemistry or in situ hybridization in endometria and placentae of cyclic and pregnant gilts, cyclic gilts injected with estrogen, and ovariectomized gilts injected with progesterone. Progesterone up-regulated SLC2A8 protein in uterine luminal (LE) and glandular epithelia during the peri-implantation period, and expression became exclusively placental, chorion and blood vessels, after Day 30. P4 up-regulated SLC2A5 mRNA in uterine LE and glandular epithelia after implantation, and the chorion expressed SLC2A5 between Days 30 and 85. AKR1B1 and SORD proteins localized to uterine LE during the peri-implantation period, but expression switched to chorion by Day 20 and was maintained through Day 85. Uterine expression of AKR1B1 mRNA was down-regulated by estrogen. KHK protein localized to trophectoderm/chorion throughout gestation. These results provide evidence that components for the conversion of glucose to fructose and for fructose transport are present at the uterine-placental interface of pigs. The shift in expression from LE to chorion during pregnancy suggests free-floating conceptuses are supported by fructose synthesized by the uterus, but after implantation, the chorion becomes self-sufficient for fructose synthesis and transport.


Assuntos
Frutose/metabolismo , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Transportador de Glucose Tipo 5/metabolismo , Placenta/metabolismo , Útero/metabolismo , Aldeído Redutase/metabolismo , Animais , Implantação do Embrião/efeitos dos fármacos , Implantação do Embrião/fisiologia , Estradiol/farmacologia , Ciclo Estral/metabolismo , Feminino , Frutose/biossíntese , L-Iditol 2-Desidrogenase/metabolismo , Ovariectomia , Placenta/efeitos dos fármacos , Gravidez , Progesterona/farmacologia , Suínos , Útero/efeitos dos fármacos
8.
BMC Genet ; 17: 50, 2016 Feb 29.
Artigo em Inglês | MEDLINE | ID: mdl-26923368

RESUMO

BACKGROUND: Reproductive efficiency has a great impact on the economic success of pork production. Gilts comprise a significant portion of breeding females and gilts that reach puberty earlier tend to stay in the herd longer and be more productive. About 10 to 30% of gilts never farrow a litter and the most common reasons for removal are anestrus and failure to conceive. Puberty in pigs is usually defined as the female's first estrus in the presence of boar stimulation. Genetic markers associated with age at puberty will allow for selection on age at puberty and traits correlated with sow lifetime productivity. RESULTS: Gilts (n = 759) with estrus detection measurements ranging from 140-240 days were genotyped using the Illumina PorcineSNP60 BeadChip and SNP were tested for significant effects with a Bayesian approach using GENSEL software. Of the available 8111 five-marker windows, 27 were found to be statistically significant with a comparison-wise error of P < 0.01. Ten QTL were highly significant at P < 0.005 level. Two QTL, one on SSC12 at 15 Mb and the other on SSC7 at 75 Mb, explained 16.87% of the total genetic variance. The most compelling candidate genes in these two regions included the growth hormone gene (GH1) on SSC12 and PRKD1 on SSC7. Several loci confirmed associations previously identified for age at puberty in the pig and loci for age at menarche in humans. CONCLUSIONS: Several of the loci identified in this study have a physiological role for the onset of puberty and a genetic basis for sexual maturation in humans. Understanding the genes involved in regulation of the onset of puberty would allow for the improvement of reproductive efficiency in swine. Because age at puberty is a predictive factor for sow longevity and lifetime productivity, but not routinely measured or selected for in commercial herds, it would be beneficial to be able to use genomic or marker-assisted selection to improve these traits.


Assuntos
Estudos de Associação Genética , Maturidade Sexual/genética , Suínos/genética , Animais , Teorema de Bayes , Cruzamento , Feminino , Marcadores Genéticos , Genótipo , Técnicas de Genotipagem , Desequilíbrio de Ligação , Masculino , Fenótipo , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Reprodução/genética , Análise de Sequência de DNA
9.
J Anim Sci Biotechnol ; 5(1): 55, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25937925

RESUMO

Litter size and birth weights are limited by uterine capacity, defined as the ability of the uterus to maintain the appropriate development of some number of conceptuses. Uterine capacity is the result of the combined effects of uterine, placental and embryo/fetal function. The number of living conceptuses that the uterus is capable of supporting is greater during early gestation compared to later gestation. Plots of log fetal weight versus log placental weight also indicate that fetal weights are less sensitive to reduced placental weight (and therefore reduced intrauterine space) in early gestation compared to late gestation. However, even in late gestation, mechanisms still exist that maintain fetal growth when the size of the placenta is reduced. One such mechanism is likely to be improved development of the folded placental-epithelial/maternal-epithelial bilayer. Fold depth, and therefore the maternal fetal interactive surface, increases as gestation advances and is greater in placenta from small fetuses. On the fetal side of the placenta, the epithelial bilayer is embedded in stromal tissue. Glycosaminoglycans are major components of stroma, including hyaluronan and heparan sulfate. Hyaluronidases and heparanases are present within placental tissues, and likely play roles in modification of stromal components to facilitate fold development. Glycosaminoglycans are polymers of forms of glucose (glucosamine, glucuronic acid, iduronic acid) suggesting that glycosaminoglycan synthesis may compete with the glucose needs of the developing fetus. Pig conceptuses are fructogenic, such that a substantial portion of glucose transferred from mother to fetus is converted to fructose. Fructose is an intermediate product in the synthesis of glucosamine from glucose, and glucosamine is linked to regulation of trophoblast cell proliferation through regulation of mTOR. These findings suggest a link between glucose, fructose, glucosamine synthesis, GAG production, and placental morphogenesis, but the details of these interactions remain unclear. In addition, recent placental epithelial transcriptome analysis identified several glucose, amino acid, lipid, vitamin, mineral and hormone transporter mechanisms within the placenta. Further elucidation of mechanisms of placental morphogenesis and solute transport could provide clues to improving nutrient transport to the pig fetus, potentially increasing litter size and piglet birth weights.

10.
Mol Reprod Dev ; 80(6): 466-73, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23585221

RESUMO

The process of placental separation is not completely understood. In domestic animals, especially cattle, it is important that expulsion of the fetal membranes takes place in a timely manner in order to achieve maximal reproductive efficiency. The activity of the matrix-metalloprotease (MMP) family of proteases is known to be reduced in placentomes from cases of retained placenta. Members of the MMP family are known to be activated by the plasminogen activator (PA) family of proteases. We hypothesized that the expression and activity of the PA family increase in the cotyledon and/or caruncle as parturition approaches, with maximal expression and activity at parturition. To test this hypothesis, we performed reverse-transcriptase quantitative PCR and plasminogen-casein zymography to detect the presence and activity of PA family members in the placentome leading up to and during parturition in spontaneous and dexamethasone-induced parturient ewes. The results from our experiments indicated that serine proteases inhibitor E1 (SERPINE1) mRNA abundance in the cotyledon was different between treatment groups (P = 0.0002). In the caruncle, gene expression for plasminogen activator urokinase-type (PLAU) was different (P = 0.0154), and there was a strong trend for differences in SERPINE1 expression (P = 0.0565). These results demonstrate that expression of the PA system in the placentome changes from late pregnancy to parturition, and the presence or activity of these enzymes may occur after fetal expulsion.


Assuntos
Parto/metabolismo , Placenta/química , Ativador de Plasminogênio Tipo Uroquinase/análise , Animais , Feminino , Perfilação da Expressão Gênica , Immunoblotting , Análise dos Mínimos Quadrados , Placenta/metabolismo , Inibidor 1 de Ativador de Plasminogênio/análise , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Gravidez , RNA Mensageiro/análise , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Carneiro Doméstico , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/metabolismo
11.
Front Genet ; 2: 20, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22303316

RESUMO

The ovary and placenta are dynamic structures requiring constant modification both structurally and through cell-cell communication capabilities. The extracellular matrix and basement membranes are primarily composed of a milieu of glycosaminoglycans, including heparan sulfate and hyaluronan. Heparanase (HPSE) and hyaluronidases (HYAL) are responsible for degrading heparan sulfate and hyaluronan, respectively. Therefore, the objective of this study was to evaluate the relationship of SNPs distinct to HPSE, HYAL1, and HYAL2 with measurements of reproduction and production traits in swine. Single trait associations were performed on a Landrace-Duroc-Yorkshire population using SNPs discovered and identified in HPSE, HYAL1, and HYAL2. Analyses were conducted on an extended pedigree and SNPs were found to be associated with reproductive and production traits. Prior to multiple-testing corrections, SNPs within HPSE were weakly associated (P < 0.03) having additive effects with age at puberty (-2.5 ± 1.08 days), ovulation rate (0.5 ± 0.24 corpora lutea), and number of piglets born alive (0.9 ± 0.44 piglets). A HYAL1 and two HYAL2 SNP were nominally associated (P ≤ 0.0063) with number of piglets born alive after multiple-testing corrections (effects between 1.02 and 1.44 piglets), while one of the same HYAL2 markers maintained a modest association (P = 0.0043) having a dominant effect with number of piglets weaned (1.2 ± 0.41 piglets) after multiple-testing correction. Functionally, HPSE and HYAL1 and 2 have been shown to participate in events related to ovarian and placental activity. SNPs from these studies could potentially assist with understanding genetic components underlying sow lifetime productivity as measured by piglet survivability based on number born alive and number weaned, thereby contributing to a greater number of pigs/sow/year.

12.
Anim Reprod Sci ; 118(2-4): 297-309, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19647381

RESUMO

The trophoblast-endometrial epithelial cell bilayer of porcine placenta undergoes microscopic folding during gestation, and the folded bilayer is embedded in fetal placental stroma. We hypothesized that hyaluronan was a component of fetal placental stroma, and that hyaluronoglucosaminidases played a role in bilayer folding. Gilts were unilaterally hysterectomized-ovariectomized (UHO) at 160 days of age, mated at estrus and killed on days 25, 45, 65, 85 or 105 of gestation. Fetal placental tissues were collected to evaluate hyaluronan and hyaluronoglucosaminidase content. Fetal placental hyaluronan concentration increased (P<0.01) between day 25 and 45 of gestation, remained high throughout gestation, and was greater (P<0.05) in the fetal placenta of the smallest compared to the largest fetuses on day 105 of gestation. Hyaluronan was localized to fetal placental stroma. Three cDNAs for hyaluronoglucosaminidase 1 (two 1379 and one 1552bp) and one cDNA (1421bp) for hyaluronoglucosaminidase 2 were cloned from day-85 fetal placental RNA. Gene expression analysis indicated that the 1379bp form of hyaluronoglucosaminidase 1 mRNA did not differ, the 1552bp form increased, and the 1421bp form of hyaluronoglucosaminidase 2 decreased during pregnancy. Amount of all three mRNAs was greater (P<0.05) in fetal placenta of the smallest compared to the largest fetuses. Zymography indicated 70 and 55kd protein isoforms of hyaluronoglucosaminidase in fetal placental tissue. Both forms increased with advancing gestation and were greater in fetal placenta of the smallest compared to the largest fetuses (P<0.05). These results are consistent with a role for hyaluronan and hyaluronoglucosaminidases in the development of the microscopic folds of the pig placenta during gestation.


Assuntos
Peso Fetal/fisiologia , Idade Gestacional , Ácido Hialurônico/análise , Hialuronoglucosaminidase/análise , Placenta/química , Placenta/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Feminino , Expressão Gênica , Hialuronoglucosaminidase/química , Hialuronoglucosaminidase/genética , Isoenzimas/análise , Isoenzimas/genética , Dados de Sequência Molecular , Tamanho do Órgão , Placenta/anatomia & histologia , Gravidez , RNA Mensageiro/análise , Alinhamento de Sequência , Suínos
13.
Reprod Fertil Dev ; 21(6): 757-72, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19567219

RESUMO

Heparanase (HPSE) is an endoglycosidase that specifically degrades heparan sulfate, which is an abundant glycosaminoglycan of the pig placenta. The aim of the present study was to clone cDNA encoding porcine HPSE and characterise the expression level and localisation of HPSE mRNA in porcine placentas throughout gestation. Placental tissues were collected from litters on Days 25, 45, 65, 85 and 105 of gestation. Three transcript variants similar to HPSE were identified in the pig placenta. In addition, the HPSE gene was mapped to pig chromosome 8 in close proximity to quantitative trait loci for litter size and prenatal survival. Real-time polymerase chain reaction and in situ hybridisation were used to characterise the expression of two HPSE variants, namely HPSE v1 and v2, in the pig placenta throughout gestation. The expression of HPSE v1 and v2 was elevated (P < 0.01) in placentas during very early gestation (Day 25) as well as during late gestation (Days 85 and 105). Finally, HPSE v1 and v2 mRNA were localised to the cuboidal trophoblast cells of the folded bilayer located nearest to the maternal endometrium. These findings illustrate that HPSE likely plays a role in the development and modification of the pig placenta, which has implications for litter size and prenatal survival.


Assuntos
Clonagem Molecular , Regulação Enzimológica da Expressão Gênica , Glucuronidase/genética , Placenta/enzimologia , RNA Mensageiro/metabolismo , Animais , Sequência de Bases , Mapeamento Cromossômico , Feminino , Idade Gestacional , Hibridização In Situ , Isoenzimas , Dados de Sequência Molecular , Placenta/citologia , Reação em Cadeia da Polimerase , Gravidez , Células Estromais/enzimologia , Suínos , Trofoblastos/enzimologia
14.
Reproduction ; 131(2): 379-88, 2006 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-16452731

RESUMO

Total protein secreted in the intrauterine lumen increases between day 10 and 13 post-estrus in both cyclic and pregnant gilts. The objective of this experiment was to identify those intrauterine proteins whose secretion changes during this time period. Sixteen mature gilts were either mated (day 0) or remained cyclic and were slaughtered at either day 10 or day 13 (n = 4 per status by day). At slaughter, each uterine horn was flushed with 20 ml Minimal Essential Medium. Flushings were dialyzed extensively against distilled water. A 0.5 ml aliquot of each was lyophilized, subjected to two-dimensional PAGE, and protein spots were identified following Coomassie staining of each gel. Densitometry was used to compare relative amounts of each spot. After statistical analysis, spots that differed due to either day, status, or day by status interaction were excised and digested in-gel with trypsin. The resulting peptides were analyzed by tandem mass spectrometry (MS/MS). Using MS/MS data, protein identification for each spot was attempted. There were 280 matching spots, of which 132 were significantly (P < 0.05 or 0.01) affected by pregnancy status, day, or the day by status interaction. Most (73%) spots increased from day 10 to day 13 with no effect of pregnancy. Several spots were identified as proteases or their inhibitors. Others potentially modify glycolipids and/or glycoproteins. These results indicate that the concentrations of many proteins within the intrauterine environment during early pregnancy are independent of the conceptus and could play roles in regulating the endometrial or conceptus glycocalyx.


Assuntos
Prenhez/metabolismo , Proteínas/análise , Suínos/metabolismo , Útero/química , Animais , Densitometria , Eletroforese em Gel Bidimensional , Feminino , Glicolipídeos/análise , Glicoproteínas/análise , Espectrometria de Massas , Peptídeo Hidrolases/análise , Gravidez , Inibidores de Proteases/análise
15.
Mol Reprod Dev ; 65(4): 366-72, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12840809

RESUMO

The porcine amphiregulin gene was previously reported to be within the quantitative trait locus (QTL) for uterine capacity on chromosome 8. Because amphiregulin stimulates cell proliferation, the amphiregulin gene might be responsible for this QTL. The objectives of this study were to clone amphiregulin cDNA and compare endometrial expression of its mRNA in pregnant Meishan (M) and White composite (WC) pigs. We obtained two amphiregulin cDNAs, one with 1,221 bp and another with 1,109 bp. The 112 bp difference corresponded to exon 5 of the human amphiregulin gene, which codes for the cytoplasmic domain. Endometrial mRNA expression of amphiregulin was significantly lower in M pigs than in WC pigs during early pregnancy (day 15 - 40 of gestation). Amphiregulin mRNA expression in the endometrium of both M and WC pigs increased (P < 0.01) from days 15 to 20, decreased (P = 0.01) from days 20 to 30, and did not change between days 30 and 40. This may result in reduced amphiregulin protein production leading to the slower development of M conceptuses, contributing to greater uterine capacity and litter size in prolific Chinese M pigs. Porcine genomic sequences isolated from a bacterial artificial chromosome genomic library contained exon 5, suggesting that the deletion of exon 5 in the mRNA may be due to differential splicing. The amphiregulin gene consisted of six exons and five introns spanning 10.3 kb. Mol.


Assuntos
Endométrio/metabolismo , Glicoproteínas/genética , Glicoproteínas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Suínos/genética , Sequência de Aminoácidos , Anfirregulina , Animais , Sequência de Bases , Cruzamento , Cromossomos Artificiais Bacterianos , Clonagem Molecular , DNA Complementar , Família de Proteínas EGF , Feminino , Expressão Gênica , Regulação da Expressão Gênica , Glicoproteínas/química , Peptídeos e Proteínas de Sinalização Intercelular/química , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Gravidez , Locos de Características Quantitativas , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Suínos/fisiologia
16.
Biol Reprod ; 68(3): 735-43, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12604620

RESUMO

Previous gene mapping analyses revealed a quantitative trait locus for uterine capacity on chromosome 8. Comparison of porcine and human genetic maps suggests that the bone morphogenetic protein receptor IB (BMPR-IB) gene may be located near this region. The objectives of this study were to 1) clone the full coding region for BMPR-IB, 2) examine BMPR-IB gene expression by the endometrium and its cellular localization in cyclic and pregnant gilts, and 3) map the BMPR-IB gene. By iterative screening of an expressed sequence tag library, we obtained a 3559-base pair cDNA clone including the full coding region of BMPR-IB. Endometrial BMPR-IB mRNA expression of White composite gilts was determined by Northern blotting in Days 10, 13, and 15 cyclic and Days 10, 13, 15, 20, 30, and 40 pregnant gilts. In cyclic gilts, endometrial BMPR-IB mRNA expression was elevated on Days 13 and 15 (P < 0.01) compared with Day 10. Expression of BMPR-IB mRNA was localized in both luminal and glandular epithelium on Day 15. However, in pregnant gilts, BMPR-IB mRNA expression was not significantly different in the endometrium from Day 10 to Day 20, and it was significantly decreased on Days 30 and 40 (P = 0.011). The BMPR-IB gene was mapped to 108 cM on chromosome 8. These findings show that BMPR-IB mRNA expression is regulated differently in cyclic and pregnant gilts; this pattern of gene expression may be important for endometrial function during the luteal phase of the estrous cycle as compared with early pregnancy.


Assuntos
Endométrio/metabolismo , Prenhez/genética , Proteínas Serina-Treonina Quinases/genética , Receptores de Fatores de Crescimento/genética , Suínos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting/veterinária , Receptores de Proteínas Morfogenéticas Ósseas Tipo I , Mapeamento Cromossômico , Clonagem Molecular , Cruzamentos Genéticos , DNA Complementar/química , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Endométrio/fisiologia , Etiquetas de Sequências Expressas , Feminino , Regulação da Expressão Gênica/fisiologia , Hibridização In Situ/veterinária , Masculino , Dados de Sequência Molecular , Mutação Puntual , Gravidez , Prenhez/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores de Fatores de Crescimento/biossíntese , Alinhamento de Sequência , Análise de Sequência de DNA , Suínos/metabolismo , Suínos/fisiologia
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