ABSTRACT: Sulfur-containing amino acids supplementation to gilts from late pregnancy to lactation altered offspring’s intestinal microbiota and plasma metabolites
Project description:16 rats were mated and the dams continued pregnancy (controls) or were subsequently caloric restricted (CR) for 20% during days 1-12. Control female/male offspring continued normal lactation, while offspring of CR-treated dams received either normal lactation (CR group) or received during lactation until PN21 leptin supplementation. Leptin treatment of offspring during lactation after caloric restriction of dams during pregnancy reverts CR-induced dysfunction.
Project description:Different lactation stages have marked influence on milk yield, milk constituents and nourishment of the neonates. However, the differential gene expression during different lactation stages in Bosindicus has not been investigated so far. In this study, we carried out high-resolution mass spectrometry-based quantitative proteomics of bovine whey at early, mid and late lactation stages of MalnadGidda (Bosindicus) cows. Using TMT-based quantitative proteomics, we compared the bovine whey proteins on progressive lactation stages of Indian breed, MalnadGidda(Bosindicus). LC-MS/MS analysis of whey peptides from early, mid and late lactation stages resulted in the generation of 420,092 MS/MS spectra and 50,800 peptide spectrum matches, which led to the identification of 4,450 peptides corresponding to 725 proteins. Out of which, 440 proteins were differentially expressed (≥1.5-fold). Gene Ontology studies showed that proteins that regulatemilk composition and mammary growth associated proteins are abundantly expressed during peak lactation stages. Whereas, proteins related to pregnancy and mammary involution are expressed high in late and mid lactation stages indicating the physiological changes in the maternal system of bovine during drying period. Detection of progestagen associated endometrial protein; an immune protein seen in the fetomaternal interface and other pregnancy associated proteins at mid lactation suggest a candidate biomarker for the early pregnancy diagnosis. These results are overlapping with the previous findings addressed in milk from exotic breeds. We strongly believe that this preliminary investigation on differential proteome in milk whey over the course of lactation of indigenous cattle could answer many unsolved questions in lactation biology.
Project description:The Chlamydomonas reinhardtii transcriptome was characterized from nutrient-replete and sulfur-depleted wild-type and snrk2.1 mutant cells; the mutant is null for the regulatory serine-threonine kinase SNRK2.1, which is required for acclimation to sulfur deprivation. The transcriptome analyses involved microarray hybridization and RNA-seq technology; RT-qPCR evaluation of the data obtained by these techniques showed that RNA-seq is significantly more quantitative than microarray hybridizations. Sulfur-deprivation-responsive transcripts included those encoding proteins involved in sulfur acquisition and assimilation, recycling of sulfur-containing amino acids, synthesis of reduced sulfur metabolites and cofactors, and modification of cellular structures such as the cell wall and complexes associated with the photosynthetic apparatus. Moreover, the data suggest that cells deprived of sulfur favors accumulation of proteins with fewer sulfur-containing amino acids. Most of these sulfur-deprivation responses are controlled by the SNRK2.1 kinase. Furthermore, the snrk2.1 mutant exhibits a set of unique responses during both sulfur-replete and sulfur-depleted conditions that are not observed in wild-type cells. Many of these responses are likely to be elicited by singlet O2 accumulation in the mutant cells. The transcriptome results for the wild-type and mutant cells strongly suggest the occurrence of massive changes in cellular physiology and metabolism as the cells become depleted for sulfur, and reveal aspects of acclimation that are likely critical for cell survival. The three supplementary files GSE17970_supplemental_table_*.xls below include results of the differential expression analysis (expression estimates, fold changes and p-values), and different clusters of functionally related genes.
Project description:Amino acid starvation during recombinant protein production (RPP) induces metabolic stress to cellular host which results in reduced productivity of the recombinant bioprocess. In present study, supplementation of amino acids in a chemically defined medium showed several folds increase in recombinant product titers in E. coli BL21 DE3 strain. To understand, the effect of amino acid supplementation in alleviating cellular stress during RPP a deep insight of cellular physiology must be studied. Here, we performed transcriptomic analysis of E. coli expressing a recombinant protein in two different conditions: with (5 mM of all 20 amino acids) as test and without supplementation of amino acids as control. RNA-seq data revealed downregulation of several genes associated with stress in test culture confirming the critical role of amino acids supplementation in improving RPP.
Project description:Using RNA-seq to explore the mechanisms underlysing the effects of methionyl-methionine dipeptide (Met-Met) or free methionine (Met) supplementation in Met-restricted mice during pregnancy on mammogenesis and lactation.
Project description:A comparison between high and low sulfur source supplies, i.e., sulfate, methionine or cystine, was carried out in order to identify key steps in the biosynthetic and catabolic pathways of the sulfur amino acids.
Project description:Developmental programming is the concept that environmental factors, particularly during foetal life, can alter development, metabolism and physiology of an organism and this can have consequences later in life. There is growing interest in developmental programming in livestock species, particularly effects of maternal pregnancy nutrition, which is easy to manipulate. Recent research, using a sheep model, has shown that milk production in ewe offspring may be susceptible to maternal nutritional programming, such that over nutrition (ad libitum) of the pregnant dam, compared with maintenance nutrition, may impair their first lactation performance and result in the weaning of lighter lambs. RNA-seq was performed to identify gene expression differences as a result of maternal nutrition in ewe offspring during their first parity. Samples were collected in late pregnancy and during lactation, allowing us to examine gene expression changes during maturation of the ovine mammary gland. Three biological replicates were sequenced for each of the treatment conditions (maternal nutrition: sub-maintenance, maintenance, and ad libitum) and time points (late pregnancy and lactation). Each biological replicate consisted of RNA from multiple individuals (late pregnancy n=3, lactation n=2).
Project description:Lipid metabolic disarray in young and adult mice offspring's liver is induced by saturated fatty acids (SFA) but prevented by alpha linolenic acid (ALA, 18:3 ω3) in the maternal diet during pregnancy and lactation. The aim of the present study was to analyse the impact of maternal dietary ALA on the liver gene expression in the new-born offspring in comparison to a SFA diet. Methods: C57Bl6/J dams were fed with diets normal in calories but rich in ALA or SFA before mating and during pregnancy. Pups were sacrificed at birth and liver parameters were assessed. Gene expression was characterized by microarray analysis and validated by real time qPCR. Results: ALA compared to SFA in maternal diets during pregnancy, increased polyunsaturated fatty acids while differentially modified fatty acid desaturase activities in offspring liver. Overall, 474 and 662 genes from born pups’ liver, were differentially regulated by ALA and SFA compared to control diet (p<0.05; Fold change 2), respectively. Notably, Per3 was up-regulated by ALA whereas down-regulated by SFA, compared to control diet. Conclusions: ALA and SFA enriched diets differentially affect gene expression pattern in the offspring’s liver. ALA in particular, upregulates genes associated to low adiposity.