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Single cell RNA-seq of ten-thousand epithelium cells from the mammary glands of mice in puberty, estrus and diestrus phases

ABSTRACT: Epithelial cells were isolated by FACS from the mammary glands of pubescent (5 week old), estrus adult (10 week old) and diestrus adult (10 week old) female mice. Freshly sorted cells were submitted to a 10X Genomics Chromium System for single cell capture. cDNA synthesis and library preparation was done according to the protocol supplied by the manufacturer. Sequencing was carried out on an Illumina NextSeq500 sequencer using parameters recommended by 10X Genomics.

ORGANISM(S): Mus musculus

PROVIDER: GSE103272 | GEO | 2017/11/20

SECONDARY ACCESSION(S): PRJNA401672

REPOSITORIES: GEO

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Single cell RNA-seq of eight thousand epithelium cells from the mammary glands of adult mice

Project description:Epithelial cells were isolated by FACS from the mammary glands of adult (10 week old) female mice. A basal subpopulation of the epithelial cells was also isolated. Freshly sorted cells were submitted to a 10X Genomics Chromium System for single cell capture. cDNA synthesis and library preparation was done according to the protocol supplied by the manufacturer. Sequencing was carried out on an Illumina NextSeq500 sequencer to achieve 75 bp paired-end reads.

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Transcriptome profiling of wildtype C57BL/6 female mice during estrous cycle(mainly in estrus and diestrus).

Project description:Background:The morphology of vaginal secretions during estrus and diestrus is characteristic and easy to distinguish. There are also significant differences in hormone levels between the two periods in physiological condition. However, the gene changes during estrus cycle especially the two periods are still unclear. Therefore, this study plans to find out the pathway changes related to the estrus cycle of female mice, the expression of various proteins, predict the genes sensitive to hormone changes in vivo through gene analysis of 15 tissues in the two periods and construct of differential gene expression library. Methods:The wildtype healthy female mice in estrus were compared with those in diestrus. The transcriptome changes between the two periods were probed by using high-throughput RNA-seq techniques. Results:A comprehensive mouse transcriptome map in estrus or diestrus across fifteen tissues including aorta, vein, brown adipose tissue, white adipose tissue, heart, kidney, liver, skeletal muscle, stomach, duodenum, cerebrum, hypothalamus, ovary, womb and spleen were provided. Conclusions:This dataset could serve as a baseline resource for investigating the potential gene changes sensitive to the estrus cycle across different tissues and provide the information for screening the key functional genes or molecular markers of estrus cycle.

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Comparative cell free salivary miRNA profile in Bubalus bubalis between diestrus and estrus stages

Project description:Despite buffaloes being primary farm animals, their reproductive performance remains poor mainly due to inaccurate estrus detection methods that ultimately has an economic impact on dairy industry as well as farmers. Recently, numerous studies showed potential of miRNAs as estrus biomarker. However, a miRNA profile of buffalo cell free saliva, a non-invasive fluid, at estrus and diestrus stages is missing. Hence, the present study was planned to identify differential levels of salivary cell free miRNAs in estrus as compared to the diestrus phase of buffalo oestrous cycle (n=3) in order to discover a possible estrus specific miRNAs as biomarkers. miRNA-Seq data analysis showed that in total 10 miRNAs i.e bta-miR-375, bta-miR-200c, bta-miR-30d, bta-let-7f, bta-miR-200a, bta-miR-12034, bta-let-7b, bta-miR-142-5p, bta-miR-2467-3p, bta-miR-30a-5p are significantly altered (log2foldchange >3 and p<0.05) during estrus in comparison to the diestrus phase in buffaloes, suggesting their estrus biomarker potential. Overall, 8 miRNAs i.e bta-miR-375 (6.87 Fold; p-value 0.003), bta-miR-200c (5.98 Fold; p-value 0.003), bta-miR-30d (4.17 Fold; p-value 0.015), bta-let-7f (3.34 Fold; p-value 0.022), bta-miR-200a (4.92 Fold; p-value 0.024), bta-miR-12034 (3.58 Fold; p-value 0.0025), bta-let-7b (3.06 Fold; p-value 0.031), bta-miR-30a-5p (4.7 Fold; p-value 0.036) were upregulated, whereas bta-miR-142-5p (-3.4 Fold; p-value 0.032) and bta-miR-2467-3p (-5.24 Fold; p-value 0.035) were downregulated during estrus. However, further validation study using qPCR is required in a large sample size in order to determine their estrus biomarker potential. In summary, our results revealed differential salivary cell free miRNAs profile during the oestrous cycle that may lead to the development of estrus specific miRNAs based point-of-care test applicable for the reproductive management of buffaloes in the field condition in the near future.

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Single-cell RNAseq (10x Genomics) analysis of mouse splenic CD4+ T cells in WT and ∆Foxp3 mice and in WT/∆Foxp3 bone marrow chimeras

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