Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Expression data of porcine mammary glands from late pregnancy to peak lactation.

ABSTRACT: Sow milk yield and quality is crucial for the survival and growth of piglets. Mammary epithelial cell proliferation, differentiation, milk component production and galactopoietic hormone and cytokine synthesis result from the quantities of differentially expressed genes. This study used the Affymetrix Porcine GeneChip to acquire a comprehensive picture of transcriptional adaptations in the mammary glands of lactating sows. Mammary tissue samples were collected from six sows at -17, 1 and 17 d relative to parturition. Mammary tissues from two sows were used to extract RNA and were subsequently pooled in equal volumes. Nine pooled samples were hybridized to porcine Affymetrix GeneChips.

ORGANISM(S): Sus scrofa

SUBMITTER: D P Shu

PROVIDER: E-GEOD-30704 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The seminal plasma (SP) is the liquid component of semen that facilitates sperm transport through the female genital tract. SP modulates the activity of the ovary, oviductal environment and uterine function during the periovulatory and early pregnancy period. Extracellular vesicles (EVs) secreted in the oviduct (oEVs) and uterus (uEVs) have been shown to influence the expression of endometrial genes that regulate fertilization and early embryo development. In some species, semen is composed of well-separated fractions that vary in concentration of spermatozoa and SP composition and volume. This study aimed to investigate the impact of different accumulative fractions of the porcine ejaculate (F1, composed of the sperm-rich fraction (SRF); F2, composed of F1 plus the intermediate fraction; F3, composed of F2 plus the post-SRF) on oEVs and uEVs protein cargo. Six days after the onset of estrus, we determined the oEVs and uEVs size and protein concentration in pregnant sows by artificial insemination (AI-sows) and in non-inseminated sows as control (C-sows). We also identified the main proteins in oEVs and uEVs, in AI-F1, AI-F2, AI-F3, and C-sows. Our results indicated that although the size of EVs is similar between AI- and C-sows, the protein concentration of both oEVs and uEVs was significantly lower in AI-sows (p < 0.05). Proteomic analysis identified 38 unique proteins in oEVs from AI-sows, mainly involved in protein stabilization, glycolytic and carbohydrate processes. The uEVs from AI-sows showed the presence of 43 unique proteins, including already-known fertility-related proteins (EZR, HSPAA901, PDS). We also demonstrated that the protein composition of oEVs and uEVs differed depending on the seminal fraction(s) inseminated (F1, F2, or F3). In conclusion, we have found a specific protein cargo in uterine and oviductal EVs depending on the type of semen fraction the sow was inseminated with, and these insemination with different seminal fractions results in the oviductal and uterine secretion of specific EVs proteins are closely associated with reproductive processes.

Project description:Once daily milking reduces milk yield and alters mammary transcriptome resulting in a decrease in milk protein synthesis as well as an induction of the apoptotic signaling networks. A local regulation due to milk stasis in the tissue could contribute to this effect but such mechanisms have not yet been described. To challenge this hypothesis, cows were milked unilaterally, once daily on one udder half and twice daily on the other one, and variations in gene expression were studied in biopsies as well as in mammary epithelial cells (MEC) shed into milk during the lactation process (milk MEC). This study therefore also contributes to decipher if transcript variations in milk purified MEC can reflect that of the mammary tissue. We compared the mammary transcript profiles in biopsies collected from unilaterally once versus twice-daily milked udder halves, 504 transcripts were differentially expressed: 193 and 232 transcripts were up- and down-regulated, respectively. A first category of transcripts, which accumulation levels are mostly up-regulated, relates to mechanisms involved in cell renewal, such as cell cycle, cellular growth and proliferation, cell death and cellular development. A second family, mostly down-regulated, is involved in small molecule biochemistry, amino acid, lipid and carbohydrate metabolisms as well as molecular transport. A third category, mostly up-regulated, includes transcripts expressed in non-epithelial mammary cells such as adipocytes, endothelial and immune cells and cells from the connective tissue. These results are consistent with previous data showing a decrease in mammary synthesis activity and an activation of cell death during once daily milking. They further suggest that during once daily milking the local milk accumulation has a major effect on mammary remodeling. Interestingly, some transcripts belonged to a third family described as M-BM-+ molecular transports M-BM-;. The expression of the 21 mRNA was then analyzed by RT-par in MEC (Table 4). Seven transcripts (NUCB2, RNASE4, ABCG2, RNASE1, SLC34A2, Cap1, FABP3, LALBA, SCD) were significantly down-regulated in milk purified MEC. Six were also significantly down-regulated in mammary biopsies. These transcripts are mostly involved in milk synthesis. We therefore conclude that milk purified MEC cannot be used as general markers of variations occurring in the mammary tissue but that variations in some transcripts listed above can be useful indicators. In this experimental design, udder halves were unilaterally milked once daily and the contralateral udder halves twice daily. Each mammary biopsy RNA sample from one udder half was compared to the one of the contralateral udder half in 5 dye-swaps corresponding to 5 animal replications, minimizing the animal variability. One dye swap corresponded to one comparison meaning two slides.

Dataset Information

Expression data of porcine mammary glands from late pregnancy to peak lactation.

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