Intracellular Calcium links Milk Stasis to Lysosome Dependent Cell Death during Early Mammary Gland Involution
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE190031 | GEO | 2022/12/01
REPOSITORIES: GEO
Similar Datasets
Project description:The molecular processes underlying human milk production and the effects of mastitic infection are largely unknown because of limitations in obtaining tissue samples. Determination of gene expression in normal lactating women would be a significant step towards understanding why some women display poor lactation outcomes. Here we demonstrate the utility of RNA obtained directly from human milk cells to detect mammary epithelial cell (MEC)-specific gene expression. Milk cell RNA was collected from 5 time points (24 hours pre-partum during the colostrum period, mid lactation, two involution, and during a bout of mastitis) in addition to an involution series comprising three time points. Gene expression profiles were determined by use of human Affymetrix arrays. Milk cells collected during milk production showed that the most highly expressed genes were involved in milk synthesis (eg. CEL, OLAH, FOLR1, BTN1A1, ARG2), while milk cells collected during involution showed a significant down regulation of milk synthesis genes and activation of involution associated genes (eg. STAT3, NF-kB, IRF5, IRF7). Milk cells collected during mastitic infection revealed regulation of a unique set of genes specific to this disease state, whilst maintaining regulation of milk synthesis genes. Use of conventional epithelial cell markers was used to determine the population of MECâ??s within each sample. This paper is the first to describe the milk cell transcriptome across the human lactation cycle and during mastitic infection, providing valuable insight into gene expression of the human mammary gland. Human milk sampling throughout lactation cycle and during mastitic infection.
2016-04-01 | E-GEOD-77978 | biostudies-arrayexpress
Project description:The molecular processes underlying human milk production and the effects of mastitic infection are largely unknown because of limitations in obtaining tissue samples. Determination of gene expression in normal lactating women would be a significant step towards understanding why some women display poor lactation outcomes. Here we demonstrate the utility of RNA obtained directly from human milk cells to detect mammary epithelial cell (MEC)-specific gene expression. Milk cell RNA was collected from 5 time points (24 hours pre-partum during the colostrum period, mid lactation, two involution, and during a bout of mastitis) in addition to an involution series comprising three time points. Gene expression profiles were determined by use of human Affymetrix arrays. Milk cells collected during milk production showed that the most highly expressed genes were involved in milk synthesis (eg. CEL, OLAH, FOLR1, BTN1A1, ARG2), while milk cells collected during involution showed a significant down regulation of milk synthesis genes and activation of involution associated genes (eg. STAT3, NF-kB, IRF5, IRF7). Milk cells collected during mastitic infection revealed regulation of a unique set of genes specific to this disease state, whilst maintaining regulation of milk synthesis genes. Use of conventional epithelial cell markers was used to determine the population of MEC’s within each sample. This paper is the first to describe the milk cell transcriptome across the human lactation cycle and during mastitic infection, providing valuable insight into gene expression of the human mammary gland.
2016-04-01 | GSE77978 | GEO
Project description:Involution returns the lactating mammary gland to a quiescent state after weaning. The mechanism of involution involves collapse of the mammary epithelial cell compartment. To test whether the cJUN NH2-terminal kinase (JNK) signal transduction pathway contributes to involution, we established mice with JNK deficiency in the mammary epithelium. We found that JNK is required for efficient involution. JNK deficiency did not alter the STAT3/5 or SMAD2/3 signaling pathways that have been previously implicated in this process. Nevertheless, JNK promotes the expression of genes that drive involution, including matrix metalloproteases, cathepsins, and BH3-only proteins. These data demonstrate that JNK has a key role in mammary gland involution post lactation.
2018-03-09 | GSE89495 | GEO
Project description:We hypothesized that accumulation of milk would influence gene expression in the mammary gland of lactating dairy cows. To test this hypothesis, we enrolled 4 multiparous Holstein cows (150 M-BM-1 10 DIM) in a half-udder milk stasis experiment. On day 1 of the experiment, cows were milked at 0430 h and at 1430 h, only right udder halves were milked. Mammary biopsies were obtained from both udder halves on day 2 of the experiment at 0500 h, immediately after right udder halves were milked and 12 h since left udder halves had last been milked. Using Affymetrix GeneChipM-BM-. Bovine Genome Arrays, we identified 32 genes that were differentially expressed between left (full) and right (empty) udder halves (fold change > 1.5; P < 0.05). Four of the genes were downregulated in response to milk stasis, whereas 28 were upregulated. Differentially expressed genes were associated with extracellular matrix remodeling, tight junction formation, regulation of blood flow, and apoptosis. In addition, four of the differentially expressed genes had been previously identified as candidates for local regulation of milk production in dairy cows. Expression of two of these candidates, early growth response-1 (EGR-1) and thrombospondin-1 (THBS-1), was validated using real-time quantitative RT-PCR. Consistent with microarray results, both genes were upregulated in response to 24-h of milk stasis (P < 0.03). Immunofluorescence was used to localize expression of EGR-1 protein, which was restricted to epithelia and was uniformly distributed. We conclude that accumulation of milk alters gene expression in the bovine mammary gland. In particular, EGR-1 and THBS-1 have emerged as strong candidates for local regulation of milk production in dairy cows. 8 samples from 4 cows
2014-01-01 | E-GEOD-27840 | biostudies-arrayexpress
Project description:Butyrophilin 1A1 (BTN1A1) was first identified as a major integral protein of both the apical surface of mammary epithelial cells during lactation and the surface membrane surrounding lipid droplets in milk. Current evidence supports a role for BTN1A1 in the secretion of milk lipids as a membrane receptor, which forms a secretion complex with the redox enzyme, xanthine oxidoreductase (XDH). Capping of cytoplasmic lipid droplets with BTN1A1 and XDH serves to envelop the droplets with cellular membranes at the cell apex as an initial step in their subsequent release from the cell. The first evidence that BTN1A1 functions in this process was the generation of Btn1a1-/- mice, in which lipid secretion was disrupted and large unstable droplets were released into alveolar spaces with fragmented surface membranes. We have revisited this mutant mouse line using RNAseq and proteomic analysis to fully assess the consequences of ablating the Btn1a1 gene on the expression of other genes and proteins. Disruption of Btn1a1 expression led to a large build-up of Xdh (6-fold over wild-type levels) in the cytoplasm, induction of acute phase response genes and Lif-activation of STAT3 phosphorylation. At peak lactation, approx. 10% of the cells were dying, as assessed by TUNEL-analysis of nuclear DNA. Cell death appeared to proceed through expression of caspase 8 and activated caspase 3 and not via lysosomal lysis, which is the principal route for elimination of cells during mammary involution in wild-type mice. Other potential death pathways include autophagy and Slc5a8-mediated inactivation of survivin (Birc5). Milk secretion was prolonged by renewal of the secretory epithelium, as evidenced by the expression of cyclins, Fos/Jun and upregulation of Ki67 in approx. 10% of secretory cell nuclei. Thus a 21-day lactation cycle can be maintained, despite significant cell death by compensatory cell division, provided a milking stimulus is maintained. These data highlight the importance of BTN1A1 expression for the maintenance of terminally differentiated mammary epithelial cells and optimal milk production throughout lactation. From a practical standpoint, the complimentary RNAseq and proteomics data should provide a useful reference database for gene and protein expression in C57/Bl6 mice at peak lactation.
2021-09-20 | PXD021249 | Pride
Project description:We hypothesized that accumulation of milk would influence gene expression in the mammary gland of lactating dairy cows. To test this hypothesis, we enrolled 4 multiparous Holstein cows (150 ± 10 DIM) in a half-udder milk stasis experiment. On day 1 of the experiment, cows were milked at 0430 h and at 1430 h, only right udder halves were milked. Mammary biopsies were obtained from both udder halves on day 2 of the experiment at 0500 h, immediately after right udder halves were milked and 12 h since left udder halves had last been milked. Using Affymetrix GeneChip® Bovine Genome Arrays, we identified 32 genes that were differentially expressed between left (full) and right (empty) udder halves (fold change > 1.5; P < 0.05). Four of the genes were downregulated in response to milk stasis, whereas 28 were upregulated. Differentially expressed genes were associated with extracellular matrix remodeling, tight junction formation, regulation of blood flow, and apoptosis. In addition, four of the differentially expressed genes had been previously identified as candidates for local regulation of milk production in dairy cows. Expression of two of these candidates, early growth response-1 (EGR-1) and thrombospondin-1 (THBS-1), was validated using real-time quantitative RT-PCR. Consistent with microarray results, both genes were upregulated in response to 24-h of milk stasis (P < 0.03). Immunofluorescence was used to localize expression of EGR-1 protein, which was restricted to epithelia and was uniformly distributed. We conclude that accumulation of milk alters gene expression in the bovine mammary gland. In particular, EGR-1 and THBS-1 have emerged as strong candidates for local regulation of milk production in dairy cows.
2014-01-01 | GSE27840 | GEO
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.
2013-08-27 | E-GEOD-43505 | biostudies-arrayexpress
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 « molecular transports ». 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.
2013-08-27 | GSE43505 | GEO
Project description:The experiment was performed on 10 lactating laboratory mice (Mus musculus, MF1). Half of the mice were shaved to increase their capacity to dissipate body heat (and thus milk production), while the other half were unshaved controls. By RNA-seq profiling of the mammary gland in shaved and unshaved lactating mice, we identified differentially expressed genes (DEGs) associated with shaving and then compared them with three sets of genes compiled from the mouse mammary gland literature, containing 1) imprinted genes, 2) milk synthesis-related genes, and 3) involution-related genes. Finally, DEGs induced by shaving were subjected to functional analysis of gene expression, with the focus on canonical pathways, upstream regulators, and downstream effects. We demonstrated that the shaving-induced increases in milk production were not associated with the changes in the expression of imprinted or milk-synthesis related genes. Instead, several lines of evidence strongly suggest that the mammary gland of shaved mice went into involution earlier than that of unshaved mice. Our interpretation of these results is that once provided with the enhanced capacity to dissipate body heat, shaved mice were able to rear their young to independence faster than unshaved mothers, benefiting potentially from shorter lactation and shorter interbirth interval to maximise their lifetime reproductive success.
2022-12-31 | E-MTAB-11654 | biostudies-arrayexpress
Project description:Post-lactational involution of the mammary gland provides a unique model to study breast cancer susceptibility and metastasis. We have shown that the short isoform of Singleminded-2 (Sim2s), a bHLH/PAS transcription factor, plays a role in promoting lactogenic differentiation, as well as in maintaining mammary epithelial differentiation and malignancy. Sim2s is dynamically expressed during mammary gland development, with expression peaking during lactation, and decreasing in early involution. To determine the role of Sim2s in involution, we used transgenic mice expressing Sim2s under the mouse mammary tumor virus (MMTV-Sim2s) promoter. Over-expression of Sim2s in the mouse mammary gland resulted in delayed involution, indicated by a lower proportion of cleaved caspase-3 positive cells and slower re-establishment of the mammary fat pad. Immunohistochemical and quantitative RNA analysis showed a decrease in apoptotic markers and inflammatory response genes, and an increase in anti-apoptotic genes, which were accompanied by inhibition of signal transducer and activator of transcription 3 (Stat3) activity. Microarray analysis confirmed that genes in the Stat3 signaling pathway were repressed by Sim2s expression, along with NFκB and other key pathways involved in mammary gland development. Multiparous MMTV-Sim2s females displayed a more differentiated phenotype compared to wild-type controls, characterized by enhanced β-casein expression and alveolar structures. Together, these results suggest a role for Sim2s in the normal involuting gland, and identify potential down-stream pathways regulated by Sim2s. Gene expression was measured in 6 mammary gland samples using CodeLink Mouse Whole Genome microarrays. 3 samples were from wild-type mice harvested at 72 hours post weaning and 3 samples were MMTV-Sim2s mice harvested at 72 hours post weaning.
2011-02-03 | E-GEOD-27012 | biostudies-arrayexpress