ABSTRACT: Microarray profiling for differential gene expression in PSMG-hCG stimulated pre-ovulatory ovarian follicles of Chinese Erhualian and Large White sows
Project description:The Chinese Erhualian is one of the most prolific pig breeds in the world, which farrows at least five more piglets per litter than Western pig breeds partly due to a greater ovulation rate. Differences in the transcriptome of Chinese Erhualian and Large White ovaries directly result in variation of ovulation rate. To understand the molecular basis related to ovulation rate in Chinese indigenous and Western breeds, samples were collected and used to hybridized. This study reveals many potential avenues of investigation for seeking new insights into ovarian physiology and the genetic control of reproduction. Expression profiling experiments were conducted to identify differentially expressed genes in ovarian follicles at the preovulatory stage of a PMSG-hCG stimulated estrous cycle from 3 Chinese Erhualian and 3 Large White cycling sows by using the Affymetrix Porcine Genechip™.
Project description:The Chinese Erhualian is one of the most prolific pig breeds in the world, which farrows at least five more piglets per litter than Western pig breeds partly due to a greater ovulation rate. Differences in the transcriptome of Chinese Erhualian and Large White ovaries directly result in variation of ovulation rate. To understand the molecular basis related to ovulation rate in Chinese indigenous and Western breeds, samples were collected and used to hybridized. This study reveals many potential avenues of investigation for seeking new insights into ovarian physiology and the genetic control of reproduction.
Project description:A CNV map in pigs could facilitate the identification of chromosomal regions that segregate for important economic and disease phenotypes. The goal of this study was to identify CNV regions (CNVRs) in pigs based on a custom array comparative genome hybridization (aCGH). We carried out a custom-made array comparative genome hybridization (aCGH) experiment in order to identify copy number variations (CNVs) in the pig genome analysing animals of diverse pig breeds (White Duroc, Yangxin, Erhualian, Tongcheng, Large White, Pietrain, Landrace and Chinese new pig line DIV ) using a tiling oligonucleotide array with ~720,000 probes designed on the pig genome (Sus scrofa genome version 9.0). In this study, a custom-made tiling oligo-nucleotide 720k array was used with a median probe spacing of 2506 bp for screening 12 pigs with a female Duroc as the reference. WD: White Duroc (♀); YX: Yangxin (♂); EH: Erhualian (♀); TC: Tongcheng (♀); LW: Large White (♀); PT: Pietrain (♂); LD1: Landrace × DIV pig 1 (♂); LD2: Landrace × DIV pig 2 (♀); DIV1: Chinese new pig line DIV 1 (♀); DIV2: Chinese new pig line DIV 2 (♀); L1: Landrace 1 (♂); L2: Landrace 2 (♂).
Project description:Erhualian (EHL) is one of the seven strains included in a Chinese indigenous pig breed called Taihu. EHL is famous for its early sexual maturity, large litter size, high adiposity, mild temper, good maternity and high tolerance to roughage and stress. To further investigate the role of miRNAs in different breed-specific metabolic characters, a comparison of miRNA expression profiles in liver of newborn Erhualian (EHL) and Large White (LW) piglets was carried out. Six LC sciences microarrays containing 236 miRNA procine probes based on miRBase Release 16.0 and 100 custom miRNA probes based on the solex deep sequencing results (li et al.,2010) were used. When compaired with LW piglets, 5 down-regulated and 8 up-regulated miRNAs were detected (fold change >1.2 and p value < 0.1) in the liver of EHL among the 236 miRbase probes. When considering the 100 custom miRNA probes, 2 down-regulated and 5 up-regulated miRNAs were observed. These results were further described in the unpublished paper Coordinated miRNA/mRNA expression profiles for understanding breed-specific metabolic characters of liver between Erhualian and Large White pigs. Six newborn male piglets were sampled from three litters (2 from each litter) of purebred EHL and LW sows and sacrificed. Liver samples were immersed in liquid nitrogen immediately after collection and then stored at -70 M-BM-0C. Total RNA was isolated using the Trizol reagent (Invitrogen, Carlsbad, CA, USA), according to the manufacturerM-bM-^@M-^Ys instructions. Total RNAs of the two littermate piglets were pooled for the microarray analysis. The pooled samples were named as EHL1-3 and LW1-3, representing 3 mixed liver RNA samples each from EHL and LW piglets, respectively. Microarray experiment was performed by a service provider (LC sciences).
Project description:Erhualian (EHL) is one of the seven strains included in a Chinese indigenous pig breed called Taihu. EHL is famous for its early sexual maturity, large litter size, high adiposity, mild temper, good maternity and high tolerance to roughage and stress. To further investigate the role of miRNAs in different breed-specific metabolic characters, a comparison of miRNA expression profiles in liver of newborn Erhualian (EHL) and Large White (LW) piglets was carried out. Six LC sciences microarrays containing 236 miRNA procine probes based on miRBase Release 16.0 and 100 custom miRNA probes based on the solex deep sequencing results (li et al.,2010) were used. When compaired with LW piglets, 5 down-regulated and 8 up-regulated miRNAs were detected (fold change >1.2 and p value < 0.1) in the liver of EHL among the 236 miRbase probes. When considering the 100 custom miRNA probes, 2 down-regulated and 5 up-regulated miRNAs were observed. These results were further described in the unpublished paper Coordinated miRNA/mRNA expression profiles for understanding breed-specific metabolic characters of liver between Erhualian and Large White pigs.
Project description:From gestation day 75 to gestation day 90, an important stage for the placental and fetal development, the fetuses grow rapidly and need adequate nutrition. The Meishan pigs and the Large White pigs employ different ways in supplying the enough nutrients and oxygen to the fetus. The Meishan pigs increased the vascular density and the Large White pigs have the second increase in the surface of placenta. To understand the molecular basis related to late gestation placenta development in Chinese indigenous and Western breeds with different placental efficiency, samples were collected and used to hybridized. The results offered new data on understanding the molecular basis of placenta efficiency, and indicated that Erhualian pigs had the more efficient than the Large White pigs. Experiment Overall Design: Placenta efficiency (the body weight of a piglet divided by the mass of its placenta) of Erhualian pigs is markedly higher than Large White pigs. so placenta samples (female) from 6 Erhuanlian pregnant gilts at gestation day 75 (E75) and day 90 (E90) and 6 Large White pregnant gilts at gestation day 75 (L75) and day 90 (L90) were collected. RNAs from two female piglet placentas from each gilt were combined to 12 pools and hybridized to the porcine Affymetrix GeneChip.
Project description:To investigate the difference between the pre-ovulatory F5 and pre-recruitment follicular stages in laying hens and identify master regulators that control the processes in these two follicular stages. RNA was isolated from a group of 3-5 pre-recruitment follicles and the most recently recruited pre-ovulatory follicle (F5). After RNA isolation, follicles underwent RNA sequencing and differential gene expression analysis occurred.
Project description:From gestation day 75 to gestation day 90, an important stage for the placental and fetal development, the fetuses grow rapidly and need adequate nutrition. The Meishan pigs and the Large White pigs employ different ways in supplying the enough nutrients and oxygen to the fetus. The Meishan pigs increased the vascular density and the Large White pigs have the second increase in the surface of placenta. To understand the molecular basis related to late gestation placenta development in Chinese indigenous and Western breeds with different placental efficiency, samples were collected and used to hybridized. The results offered new data on understanding the molecular basis of placenta efficiency, and indicated that Erhualian pigs had the more efficient than the Large White pigs.
Project description:Experiments were designed to evaluate changes in the transcriptome (mRNA levels) in the ovulatory, luteinizing follicle of rhesus monkeys, using a controlled ovulation (COv) model that permits analysis of the naturally selected, dominant follicle at specific intervals (0, 12, 24, 36 hours) after exposure to an ovulatory (exogenous hCG) stimulus during the menstrual cycle. Total RNA was prepared from individual follicles (n=4-8/timepoint), with an aliquot used for microarray analysis (AffymetrixTM Rhesus Macaque Genome Array) and the remainder applied to quantitative real-time PCR (q-PCR) assays. The microarray data from individual samples distinctly clustered according to timepoints, and ovulated follicles displayed markedly different expression patterns from unruptured follicles at 36 h. Between timepoint comparisons revealed profound changes in mRNA expression profiles. The dynamic pattern of mRNA expression for steroidogenic enzymes (CYP17A, CYP19A, HSD3B2, HSD11B1, HSD11B2), StAR, and gonadotropin receptors (LHCGR, FSHR) as determined by microarray analysis correlated precisely with those from blinded q-PCR assays. Patterns of mRNA expression for EGF-like factors (AREG, EREG) and processes (HAS2, TNFAIP6) implicated in cumulus-oocyte maturation/expansion were also comparable between assays. Thus, several mRNAs displayed the expected expression pattern for purported theca (e.g., CYP17A, AREG), granulosa (CYP19A, FSHR), cumulus (HAS2, TNFAIP6) cell, and surface epithelium (HSD11B) related genes in the rodent/primate preovulatory follicle. This database will be of great value in analyzing molecular and cellular pathways associated with periovulatory events in the primate follicle (e.g. follicle rupture, luteinization, inflammatory response, and angiogenesis), and for identifying novel gene products controlling mammalian fertility. Keywords: time course Total RNA was prepared from individual follicles (n=4-8/timepoint). Dominant follicles were selected specific intervals (0, 12, 24, 36 hours) after exposure to an ovulatory (exogenous hCG) stimulus during the menstrual cycle.