Project description:Previously we have shown significant differences in lactation performance, mammary gland histology and expression profiles of mammary transcriptome during peak-lactation (lactation day 9; L9) between the ordinary CBA/CaH (CBA) and the superior QSi5 strains of mice. In the present study, we compared mammary gland histology between CBA and QSi5 at mid-pregnancy (pregnancy day 12; P12). We assessed lactation performance during the first 8 days of lactation of the 13th - 14th generation of the Advanced Intercross Line (AIL) (CBA X QSi5) mice. We utilized an integrative approach to analyzing mammary microarray expression profiles of CBA and QSi5 at P12 and CBA, AIL and QSi5 at L9. The inguinal mammary glands of CBA/CaH and QSi5 during mid-pregnancy (Pregnancy day 12; P12), and the glands of CBA/CaH, AIL and QSi5 during peak lactation (Lactation day 9; L9) were collected and total RNA was extracted for Affymetrix microarray (mouse genome 430 2) assay
Project description:The mammary gland of the lactating mouse synthesizes and secretes milk lipid equivalent to its entire body weight in a single 20 day lactation cycle, making it one of the most active lipid synthetic organs known. To test the hypothesis that multiple metabolic control points and potential regulatory mechanisms are involved in activating lipid and lactose synthesis at the onset of lactation we compared the mammary transcriptome of 130 genes involved in glucose metabolism between late pregnancy and early lactation and in response to dietary fat. We utilized data obtained from microarray analysis of mammary glands from quadruplicate FVB mice at pregnancy day 17, and lactation day 2. Diets containing 8% or 40% lipid were fed from lactation days 5 to 10 and mammary glands and livers of triplicate FVB mice prepared for microarray analysis. We also compared the metabolome obtained from magnetic resonance spectroscopy of flash frozen glands of the mammary gland at day 17 of pregnancy with that at day 2 of lactation. The results provide a global picture of the multiple metabolic strategies utilized to turn a quiescent organ into an incredibly efficient machine for massive but balanced lipid and lactose synthesis and implicate the transcription factor SREBP-1c in regulation of part of the pathway. Keywords: Timecourse and dose response to dietary fat
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 mammary gland of the lactating mouse synthesizes and secretes milk lipid equivalent to its entire body weight in a single 20 day lactation cycle, making it one of the most active lipid synthetic organs known. To test the hypothesis that multiple metabolic control points and potential regulatory mechanisms are involved in activating lipid and lactose synthesis at the onset of lactation we compared the mammary transcriptome of 130 genes involved in glucose metabolism between late pregnancy and early lactation and in response to dietary fat. We utilized data obtained from microarray analysis of mammary glands from quadruplicate FVB mice at pregnancy day 17, and lactation day 2. Diets containing 8% or 40% lipid were fed from lactation days 5 to 10 and mammary glands and livers of triplicate FVB mice prepared for microarray analysis. We also compared the metabolome obtained from magnetic resonance spectroscopy of flash frozen glands of the mammary gland at day 17 of pregnancy with that at day 2 of lactation. The results provide a global picture of the multiple metabolic strategies utilized to turn a quiescent organ into an incredibly efficient machine for massive but balanced lipid and lactose synthesis and implicate the transcription factor SREBP-1c in regulation of part of the pathway. FVB control time-series mice were on normal lab chow diets and mammary tissue was taken at Pregnant days 12 and 17, plus lactation days 1, 2, and 9 in those animals. the time-points were taken as quadruplicates for each developmental stage. for the dietary component, triplicate FVB dams were fed normal lab chow diets until lactation day 5, when they were converted to diets containing either 8% or 40% Kcal from soy oils (see supplementary table to publication for further detail) for 5 days. At lactation day 10, mammary and liver tissues were taken from these animals and preparred for microarray analysis. All tissues were stabilized for RNA content in RNALater and were then isolated and cleaned up following manufacturer's instructions (Qiagen).
Project description:Cross-species hybridization analysis of mammary glands during pregnancy and lactation. Results provide insight into putative conserved molecular mechanisms regulating mammary gland development. This study was performed to identify orthologous transcripts that are differentially co-expressed in the mammary gland at 2 stages of development (pregnancy and lactation) in wild type Sprague-Dawley rats.
Project description:Previously we have shown significant differences in lactation performance, mammary gland histology and expression profiles of mammary transcriptome during peak-lactation (lactation day 9; L9) between the ordinary CBA/CaH (CBA) and the superior QSi5 strains of mice. In the present study, we compared mammary gland histology between CBA and QSi5 at mid-pregnancy (pregnancy day 12; P12). We assessed lactation performance during the first 8 days of lactation of the 13th - 14th generation of the Advanced Intercross Line (AIL) (CBA X QSi5) mice. We utilized an integrative approach to analyzing mammary microarray expression profiles of CBA and QSi5 at P12 and CBA, AIL and QSi5 at L9.
Project description:Cross-species hybridization analysis of mammary glands during pregnancy and lactation. Results provide insight into putative conserved molecular mechanisms regulating mammary gland development. This study was performed to identify orthologous transcripts that are differentially co-expressed in the mammary gland at 2 stages of development (pregnancy and lactation) in wild type Sprague-Dawley rats. Key points are examined in a time series of Sprague Dawley rat mammary gland development, secretory activation and lactation. Triplicate rat (three biological replicates) at each time point were used for statistical power totalling 12 individual arrays in this study. Rats were as staged pregnant day 1 the day that post coital plug was observed, and similarly, lactation day 1 was the first day after birth. Whole mammary glands No. 4 (inguinal) were obtained from female rats at stages of development: virgin (adulthood, 14 wks of age), Pregnant (5 and 14 days of pregnancy) and Lactating (day 1 and 12 postpartum). The two-color (Cy5/Cy3) microarray experiment was designed to hybridize samples from each group against a common reference, a pool of RNA from mammary gland of three parous or virgin female rats.