Vaginal swab lipidome profiles at 48 h reflect the fat composition of neonatal diet during first two days postnatal
Ontology highlight
ABSTRACT:
ORGANISM(S): Sus Scrofa Pig
TISSUE(S): Vaginal Epithelium
SUBMITTER: 
Christina Ferreira
PROVIDER: ST001224 | MetabolomicsWorkbench | Thu Jun 06 00:00:00 BST 2019
REPOSITORIES: MetabolomicsWorkbench
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Project description:Inborn errors of lipid metabolism illustrate the importance of proper milk fat oxidation in newborn mammals. In the liver, a remarkable lipid catabolic competence is present at birth; however, it is unclear how this critical trait is acquired and regulated. In this work, we found that the genes required for milk lipid catabolism are already transcribed before birth in the term fetus (E19.5) and controlled by the peroxisome-proliferator activated receptor alpha (PPARα) in mouse liver. The developmental activity of PPARα strongly regulates fatty acid oxidation genes. Two days after birth (P2), during milk suckling, PPARα-null mice develop a congenital steatosis and milk protein oxidation is de-repressed to fuel an alternative energy pathway that maintains glucose homeostasis and postnatal growth. Our results demonstrate for the first time, the developmental role of PPARα in regulating the metabolic ability to use maternal milk as fuel in the early days of life.
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Project description:Inborn errors of lipid metabolism illustrate the importance of proper milk fat oxidation in newborn mammals. In the liver, a remarkable lipid catabolic competence is present at birth; however, it is unclear how this critical trait is acquired and regulated. In this work, we found that the genes required for milk lipid catabolism are already transcribed before birth in the term fetus (E19.5) and controlled by the peroxisome-proliferator activated receptor alpha (PPARα) in mouse liver. The developmental activity of PPARα strongly regulates fatty acid oxidation genes. Two days after birth (P2), during milk suckling, PPARα-null mice develop a congenital steatosis and milk protein oxidation is de-repressed to fuel an alternative energy pathway that maintains glucose homeostasis and postnatal growth. Our results demonstrate for the first time, the developmental role of PPARα in regulating the metabolic ability to use maternal milk as fuel in the early days of life. Expression profile difference between PPARalpha wild-type (3 males and 3 females) and knock-out (3 males and 3 females) mouse liver in suckling mice. Two days after birth, pups were killed and their liver promptly removed and frozen in liquid nitrogen.
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Project description:The 4 day gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes. Serial milk samples were collected every 3h for 4 days from 5 normal, lactating women. RNA was isolated from the milk fat globules and utilized for microarray analyses.
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Project description:The 4 day gene expression profile in the lactating mammary gland was demonstrated using the Human Ref-8 BeadChip array (Illumina, Inc). Of the 22,184 gene transcripts on the array, 14,070 genes were consistently expressed and represented the milk fat globule transcriptome. Milk protein genes were among the most highly expressed along with genes involved in the milk syntesis processes.
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Project description:Studies of normal human mammary gland development and function have mostly relied on cell culture, limited surgical specimens, and rodent models. Although RNA extracted from human milk has been used to assay the mammary transcriptome non-invasively, the transcriptome derived from the milk fat layer has not been compared with the mammary-derived transcriptome nor have sources of RNA been quantified in milk. In this study the effects of milk collection and processing on RNA quality and origin were assessed in humans and rhesus macaques. Total RNA in milk was quantitated in acridine orange-stained milk using an automated whole slide scanner and custom-built Globulator software. Total RNA extracted from milk fat, cells in milk, and mammary biopsies of lactating rhesus macaques were compared using RNA sequencing and analysis. Compared with human milk, milk from macaques contained similar amounts of RNA-containing cytoplasmic crescents, but more cells. Total RNA extracted from milk fractions was also evaluated for factors that affect RNA quality. Degradation of RNA extracted from human milk fat was positively correlated with geographic distance from collection site, storage time, and sample type. There were no differences in RNA degradation in macaque milk collected after 10 min or 4 hr accumulation, suggesting that degradation of RNA extracted from milk fat may not occur in the mammary gland. Using RNA-Seq, RNA extracted from macaque milk fat and cells in milk more accurately represented RNA from mammary epithelial cells (cells that produce milk) than did RNA from mammary tissue. Mammary epithelium-specific transcripts were more abundant in macaque milk fat whereas adipose or stroma-specific transcripts were more abundant in mammary tissue. Functional analyses confirmed the validity of milk as a source of RNA from mammary epithelial cells. Analysis of highly abundant putative microRNAs in macaque milk fat revealed a potentially novel non-coding RNA species that is conserved in humans. RNA extracted from the milk fat during lactation accurately portrayed the RNA profile of milk-producing mammary epithelial cells. However, this sample type clearly requires protocols that minimize RNA degradation.
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Project description:Studies of normal human mammary gland development and function have mostly relied on cell culture, limited surgical specimens, and rodent models. Although RNA extracted from human milk has been used to assay the mammary transcriptome non-invasively, the transcriptome derived from the milk fat layer has not been compared with the mammary-derived transcriptome nor have sources of RNA been quantified in milk. In this study the effects of milk collection and processing on RNA quality and origin were assessed in humans and rhesus macaques. Total RNA in milk was quantitated in acridine orange-stained milk using an automated whole slide scanner and custom-built Globulator software. Total RNA extracted from milk fat, cells in milk, and mammary biopsies of lactating rhesus macaques were compared using RNA sequencing and analysis. Compared with human milk, milk from macaques contained similar amounts of RNA-containing cytoplasmic crescents, but more cells. Total RNA extracted from milk fractions was also evaluated for factors that affect RNA quality. Degradation of RNA extracted from human milk fat was positively correlated with geographic distance from collection site, storage time, and sample type. There were no differences in RNA degradation in macaque milk collected after 10 min or 4 hr accumulation, suggesting that degradation of RNA extracted from milk fat may not occur in the mammary gland. Using RNA-Seq, RNA extracted from macaque milk fat and cells in milk more accurately represented RNA from mammary epithelial cells (cells that produce milk) than did RNA from mammary tissue. Mammary epithelium-specific transcripts were more abundant in macaque milk fat whereas adipose or stroma-specific transcripts were more abundant in mammary tissue. Functional analyses confirmed the validity of milk as a source of RNA from mammary epithelial cells. Analysis of highly abundant putative microRNAs in macaque milk fat revealed a potentially novel non-coding RNA species that is conserved in humans. RNA extracted from the milk fat during lactation accurately portrayed the RNA profile of milk-producing mammary epithelial cells. However, this sample type clearly requires protocols that minimize RNA degradation. Transcript profiles from milk cells, milk fat, and mammary tissue from 6 lactating rhesus macaques at 30 and 90 days lactation; 34 samples run in triplicate
2013-12-12 | E-GEOD-49765 | biostudies-arrayexpress
Project description:In order to gain insight into hepatic metabolic pathways and key transcripts affecting traits related to body composition we aimed to compare samples of pigs of two breeds, the âobeseâ German Landrace (DL) and the âleanâ Pietrain (Pi) obtained at prenatal stages (35, 63, and 91 days post conceptionem) and at adult age (180 days). In terms of number of genes regulated the most striking differences between DL and Pi were found at adult age with upregulation of key genes of lipid biosynthesis/metabolism pathways (FASN, ACSS2, ACACA) in obese DL pigs on the one hand and upregulation of genes of cell growth and/or maintenance, protein syntheses as well as cell proliferation pathways (PPARD, POU1F1, IGF2R) in lean Pi pigs on the other hand. Time course analysis of expression profiles of breed differences from foetal to adult stage and functional cluster analysis of the biological processes confirmed the trend of differences between genetically different obese and lean breeds. The highlighted classes of genes showed common breed-typical expression throughout prenatal development and at adult age. The transcriptional differences between obese and lean pigs involving lipid pathways and cell activity are already initiated during early prenatal development. The information about genetic differences between obese and lean pigs reveals a number of functional candidate genes for traits related to obesity and leaness. Porcine liver from three prenatal stages (35, 63, and 91 day post conceptionem, dpc) plus adult age (180 days) of 10 animals of each of the breeds DL and Pi, which differ in body composition, were collected. The hepatic gene expression patterns of DL and Pi breeds were compared at each of the four stages of development. Three or four biological replicate microarray hybridisations were performed for each stage with DL pools labelled with Cy5 and Pi pools labelled with Cy3.
2010-06-30 | E-GEOD-5399 | biostudies-arrayexpress
Project description:In order to gain insight into hepatic metabolic pathways and key transcripts affecting traits related to body composition we aimed to compare samples of pigs of two breeds, the “obese” German Landrace (DL) and the “lean” Pietrain (Pi) obtained at prenatal stages (35, 63, and 91 days post conceptionem) and at adult age (180 days). In terms of number of genes regulated the most striking differences between DL and Pi were found at adult age with upregulation of key genes of lipid biosynthesis/metabolism pathways (FASN, ACSS2, ACACA) in obese DL pigs on the one hand and upregulation of genes of cell growth and/or maintenance, protein syntheses as well as cell proliferation pathways (PPARD, POU1F1, IGF2R) in lean Pi pigs on the other hand. Time course analysis of expression profiles of breed differences from foetal to adult stage and functional cluster analysis of the biological processes confirmed the trend of differences between genetically different obese and lean breeds. The highlighted classes of genes showed common breed-typical expression throughout prenatal development and at adult age. The transcriptional differences between obese and lean pigs involving lipid pathways and cell activity are already initiated during early prenatal development. The information about genetic differences between obese and lean pigs reveals a number of functional candidate genes for traits related to obesity and leaness. Keywords: pig, oligo-microarray, prenatal liver expression, adult liver expression, obesity, leaness
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Project description:Mammary gland (MG) de novo lipogenesis contributes significantly to milk fat in animals but little is known in humans. We hypothesized that de novo lipogenesis, as reflected by incorporation of 13C carbons from [U-13C]glucose into fatty acids (FAs) and glycerol in triglycerides (TG), will be greater: a. in milk than plasma TG, b. during a high carbohydrate (H-CHO) diet than high fat (H-FAT) diet and c. during feeding than fasting. Healthy lactating women were studied on two isocaloric, isonitrogenous diets. On one occasion subjects received diets containing H-FAT or H-CHO diet for 1 week. Incorporation of 13C from infused [U-13C]glucose into FAs and glycerol was measured using GC/MS methodology and gene expression using RNA isolated from breast milk fat globule (MFG). Incorporation of 13C2 into milk FAs, increased with increased chain length of the FAs from C2:0 to C12:0 but progressively declined in C14:0 and C16:0 and was not detected in FAs >C16. During feeding, regardless of diets, enrichment of 13C2 in milk FA and 13C3 in milk glycerol were ~3 and ~7 fold higher compared to plasma FA and glycerol, respectively. Following an overnight fast during H-CHO and H-FAT diets study periods, 25% and 6%, respectively, of medium chain FAs (MCFAs, C6-C12) were derived from glucose but increased to 75% and 25% with feeding. The expression of genes involved in FA or glycerol synthesis pathways was unchanged regardless of diet or fast-fed conditions. Conclusions: The human MG is capable of de novo lipogenesis, of primarily MCFAs and glycerol, which is influenced by the macro-nutrient composition of the maternal diet. On day 7 of consumption of each of the two diets milk samples collected from 7 healthy, lean, exclusively breastfeeding women following an overnight fast and feeding conditions [7 x 2 x 2 = 28 samples] were processed for isotope enrichments and RNA isolation from the milk fat globules. The total RNA was utilized for microarray analyses.
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