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Series GSE107419 Query DataSets for GSE107419
Status Public on Feb 09, 2018
Title Comprehensive mapping of transcriptional networks specifying lactogenic differentiation of murine mammary epithelial stem like cells
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Purpose: Understanding of gene transcriptional networks specifying murine mammary epithelial cells (HC11) during lactogenic differentiation and its comparision with murine embryonic stem cells
Methods: RNA-sequencing after ribosomal RNA depletion followed by cDNA synthesis, adaptor ligation and sequencing in duplicate of each sample using Illumina Hi-seq 2500 paired end sequencing. Sequence reads with adapters, over-represented and low-quality reads were discarded. High quality reads were mapped to reference mouse genome (mm10) using Bowtie followed derivation of FPKM values using cufflinks and DE-seq to observe differentially expressed genes.
Results: Using standard analysis methods which were published recently we uniquely mapped more than 50 million sequence reads out of which total 11961 transcripts were from ESC, 11045 from Normal, 11243 from Primed and 10719 from prolactin stage. When we did DE-seq differential gene expression analysis we observed that for Normal to primed transition 1141 and 1026 transcripts were Up and Down regulated respectively where as upon transition from Primed to Prolactin stage 385 and 655 transcripts were Up and Down regulated respectively. We also did pathway analysis to interpret the transcript activation and its function during lactogeinc differentiation; we found that the pathways which are highly up regulated are related to carbohydrate, lipids, nucleic acid and protein metabolism. We also observed that most of the up regulated pathways were related to maintenance of mammary cells differentiated state and nutrition of the infants. Our study concludes that overall differentiation requires priming with glucocorticoids followed by prolactin stimulation of HC11 cells, which tunes the cells for the secretion of milk and its maintenance of the differentiated state till the lactation period.
Conclusions: Transcriptome analysis by next generation sequencing allowed us to explore global gene expression during lactogenic differentiation of HC11 cells in presence of glucocortiocids in primed state. Glucocorticoids are activating genes, transcription factors, epigenetic modifiers and metabolic pathways which are required for later stages of lactation. On other hand gene expression analysis of prolactin stage revealed that there is a focused gene expression exclusively related to lactation process. We also observed that HC11 stem like marker are highly expressed in undifferentiated state whose expression was down regulated in primed and prolactin state indicating that HC11 are partially bipotent cells; when these cells receive differentiation signals they loose their stem cell like characteristics and are differentiated.
 
Overall design Paired end sequencing of transcriptome in replicates of embyonic stem cells, normal, primed and prolactin. Analysis of data and validation by real-time pcr analysis.
Web link http://doi: 10.1186/s13104-018-3351-4
 
Contributor(s) Sornapudi T, Nayak R, Uppada V, Guthikonda PK, Kethavath S, Yellaboina S, Pasupulati AK, Kurukuti S
Citation(s) 29642945, 30082875
Submission date Nov 28, 2017
Last update date Jun 14, 2019
Contact name Sreenivasulu Kurukuti
E-mail(s) skurukuti@gmail.com
Phone 040-66794581
Organization name University of Hyderabad
Department Animal Biology
Lab Journey to
Street address Room No-S60, School of Life Sciences, University of Hyderabad, Gachibowli
City Hyderabad
State/province Telangana
ZIP/Postal code 500046
Country India
 
Platforms (1)
GPL17021 Illumina HiSeq 2500 (Mus musculus)
Samples (8)
GSM2866581 Embryonic Stem cells Rep 1
GSM2866582 Embryonic Stem cells Rep 2
GSM2866583 Normal HC11 Rep 1 (N)
Relations
BioProject PRJNA420041
SRA SRP125747

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Supplementary file Size Download File type/resource
GSE107419_DEG_N_vs_ESC_DEseq.xlsx 6.2 Mb (ftp)(http) XLSX
GSE107419_DEG_P_vs_N_DEseq.xlsx 1.8 Mb (ftp)(http) XLSX
GSE107419_DEG_Prl_vs_N_DEseq.xlsx 1.9 Mb (ftp)(http) XLSX
GSE107419_DEG_Prl_vs_P_DEseq.xlsx 1.8 Mb (ftp)(http) XLSX
GSE107419_FPKM_ESC_N_P_PRL.xlsx 2.3 Mb (ftp)(http) XLSX
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Processed data are available on Series record

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