The gene expression program of the developing mammary gland
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE2988 | GEO | 2006/05/22
SECONDARY ACCESSION(S): PRJNA91939
REPOSITORIES: GEO
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Project description:Background: Heterogeneity within the mouse mammary epithelium has been recently explored by single-cell RNA profiling. To further understand how cellular diversity changes during mammary development, we profiled single cells from different developmental stages including late embryogenesis, early postnatal, prepuberty, resting adult, mid-pregnancy, late-pregnancy, lactation and post-involution. We also compared the transcriptomes and chromatin accessibility of micro-dissected ducts versus terminal endbuds (TEBs) to help decipher the molecular properties of these specialized units that drive morphogenesis. Results: The single cell transcriptomes of 110,726 cells isolated from different stages were determined on the 10x Chromium platform and integrative analyses were performed to compare specific time-points. The mammary rudiment at E18.5 aligned more closely with the basal than the luminal lineage, while prepubertal epithelial cells at 2 weeks exhibited lineage segregation but to a less differentiated state than their adult counterparts. Integrated analysis of five stages spanning the pregnancy cycle revealed a novel alveolar progenitor that arises in late-pregnancy and stage-specific expression profiles. A continuum of states was uncovered along the basal-luminal progenitor cell axis following interrogation with >3,200 lineage-specific signature genes. Finally, exploration of micro-dissected TEBs and ducts showed that luminal cells within TEBs harbored intermediate expression profiles, and that ductal basal cells exhibited a higher level of accessible chromatin than their counterparts in TEBs.
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Project description:The terminal end buds (TEB) are the growing part of the ductal mammary epithelium during puberty, which enable the formation of the primary mammary epithelial network. These bulbous end structures are highly proliferative, and this allows the ductal expansion into the mammary fat pad. The TEB comprise of an outer cap cell layer, which include the progenitor cells of the myoepithelium, and the body cells, which are thought to be the progenitors from which the luminal epithelium is formed. Since TEBs make up only a minute part of the whole mammary tissue, associated factors can be easily missed in whole-tissue section analysis. We have therefore optimised a method to enzymatically separate TEB and ducts, respectively, from the surrounding stroma of pubertal mice and performed transcriptome analysis on the isolated structures to identify potential novel regulators of epithelial outgrowth.
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Project description:7,12-Dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinoma is a well-recognized model, however, the genetic alterations during its carcinogenesis have yet to be determined. We used laser capture microdissection to specifically isolate cells from terminal end buds (TEBs), the origin of carcinoma, at 2 weeks after sesame oil- treatment (control) or DMBA-treatment (DMBA-TEBs), ductal carcinoma in situ (DCIS) and invasive mammary carcinoma (MC). Using an oligonucleotide microarray representing 20600 rat probe sequences, we analyzed gene expression profiles and validated mRNA and protein levels of genes of interest by real-time quantitative PCR, immunohistochemistry and immunofluorescence. The number of differentially expressed genes was smallest in DMBA-TEBs (63), followed by DCIS (798) and MC (981). Only the expression of PEP-19, an anti-apoptic gene, showed significant increases in DMBA-TEBs (4-fold), DCIS (10-fold) and MC (16-fold). MMP-13 expression was increased markedly in DCIS (19-fold) and MC (61-fold) while OPN expression was increased 6-fold in DCIS and 8-fold in MC. MMP-7 expression was increased 4-fold in MC. Nidogen-1; a participant in the assembly of basement membranes, TSP-2; an inhibitor of angiogenesis and COUP-TFI; a transcription repressor showed significant decreases in DCIS (4-, 9-, and 17-fold, respectively) and MC (10-, 37-, and 100-fold). Network analyses with IPA software revealed that the most significant network was centered on Akt groups in DCIS and ERK groups in MC. The present findings provide insights into the molecular changes and suggest the importance of PEP-19 overexpression very early on during mammary carcinogenesis. Keywords: Comparative experiments of sesame oil treatmentM-oM-<M-^HcontrolM-oM-<M-^Iand 7,12-Dimethylbenz[a]anthraceneM-oM-<M-^HDMBAM-oM-<M-^Itreatment group. Or comparative experiments of tissue type ExperimentM-oM-<M-^Z7,12-Dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinoma is a well-recognized model, however, the genetic alterations during carcinogenesis have yet to be determined. We used laser capture microdissection to specifically isolate the cells from terminal end buds (TEBs), the origin of MC, at 2 weeks after sesame oil-(control) or DMBA-treatment (DMBA-TEBs), ductal carcinoma in situ (DCIS) and invasive mammary carcinoma (MC). At 50 days after birth, inbred Sprague-Dawley female rats were given 1ml sesame oil (control) or 20 mg DMBA (Nacalai tesque, Inc. Kyoto Japan) dissolved in 1ml sesame oil, by gastric intubations. Animals in control group were euthanized after 2 weeks (W) and DMBA-treated group were euthanized after 2 weeks (W), 6W and 8W, respectively. From for 4 sample(control, DMBA-TEBs, DCIS, MC), we performed a hybridization repeatedly, respectively.
2008-10-28 | E-GEOD-13356 | biostudies-arrayexpress
Project description:To identify candidate genes involved in maturation and flowering, we conducted microarray expression studies using two poplar genotypes (Populus trichocarpa x P. deltoides hybrids) represented in continuous age gradients of one to six years. We designed 70-mers for 228 poplar genes and microarray studies were carried out using the microplate-based 96-well BioGridArray platform (GeneXP Biosciences). Floral buds, vegetative buds and shoot tips were collected at different seasonal time points from juvenile and adult trees and from both basal and upper branches of mature trees. The experiment was carried out using the microplate-based 96-well BioGridArray platform (GeneXP Biosciences). Each of 228 oligonucletides were duplicated in each well. Two human genes, beta-actin and gapdh , were also printed on all arrays as negative controls. plus ten arabidopsis oligonucleotides selected by GeneXP and used to measure quality of hybridization. For each of 16 samples, two seperate RNA isolations were performed and were considered as biological replicates. Each biological replicate was labeled with Cy5, and hybridized to duplicated wells.
2010-07-01 | E-GEOD-3457 | biostudies-arrayexpress
Project description:Diethlstilbestrol (DES) is a synthetic estrogen prescribed to several millions of pregnant women worldwide. The risk for breast cancer after age 40 in women prenatally exposed to DES has been reported, however, the precise mechanism of susceptibilities to breast cancer remains to be resolved. To understand the mechanism of effect of intrauterine exposure to DES on human mammary glands at peripubertal stage, we investigated the global gene expression profile of terminal end buds (TEBs), the target of carcinogen, in rat mammary glamds at postnatal days (PND) 35 and 49, equivalent to peripubertal stage in human, exposed to low or high dose of DES at neonatal period. In all group, the number of TEBs was gradually increased and peaked at PND35 and decreased at PND49. At PND 35 and 49, low dose of DES-treated group showed highest number of TEBs. At PND35, β- and γ-casein mRNA expression inreased 8.19-fold and 26.05-fold, respectively, and the most significant network revealed by IPA analyses showed relevance of NF-κB in low dose of DES-treated group. The present findings suggested that deregulation of mammary gland differentiation and development-related genes group may contribute to the increase of number of TEBs at critical period of carcinogen susceptibilities. Experiment: We used laser capture microdissection to specifically isolate the cells from terminal end buds (TEBs) in rat mammary gland at 35 and 49 days after birth. Inbred Sprague-Dawley female rats were subcutaneously given 0.05ml sesame oil (group I, control) and either 1 μg (group II) or 100 μg (group III) diethlstilbestrol (DES, Sigma Chemical Co., St. Louis, Mo. USA) dissolved in 0.05 ml sesame oil within 24 hr of birth. Animal were weaned at 21 days of birth and each group of rats was autopsied at 35 and 49 days of birth. From group I (35 days of birth: n=2, 49 days of birth: n=2), group II (35 days of birth: n=2, 49 days of birth: n=2), group III (35 days of birth: n=2, 49 days of birth: n=2), we performed a hybridization, respectively.
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