RESUMEN
SATB homeobox 1 (SATB1) and its heterodimeric partner SATB2 play an important regulatory role in maintaining proliferation of trophoblast stem (TS) cells and in inhibiting trophoblast differentiation. To identify the SATB-regulated genes in TS cells, we studied the transcriptome changes in a 'loss of function' model of Rcho-1 rat TS cell line. Satb1 gene expression was silenced by lentiviral delivery of shRNAs targeted to exon 9 and exon 12. An Egfp shRNA was used as a non-targeted control. Total RNA was purified from shRNA-transduced Rcho-1 cells, and whole transcriptome was assessed by RNA-sequencing on an Illumina HiSeq X platform. Differentially expressed genes in Satb1 shRNA-transduced cells were identified by analyses of the RNA-sequencing data using CLC Genomics Workbench. Differentially expressed genes with each of the two different shRNAs were compared to identify SATB1-target genes and to eliminate the potential off-targets of the shRNAs. These datasets can be used to identify the SATB-regulated genes in TS cells and to understand the molecular mechanisms that regulate trophoblast proliferation and inhibit differentiation.
RESUMEN
Disruption of estrogen receptor beta (ESR2) dysregulates oocyte maturation, which leads to failure of ovulation. We investigated ESR2-regulated genes during gonadotropin-induced oocyte maturation using RNA-sequencing. Through the administration of pregnant mare's serum gonadotropin (PMSG), synchronized follicle development was initiated in four-week-old wildtype and Esr2-null female rats. Forty-eight hours after the PMSG injection, human chorionic gonadotropin (hCG) was used for further maturation. Oocytes were collected from the ovaries 4 h after hCG injection. The total RNA was isolated from the oocytes and the whole oocyte transcriptome was determined by RNA-sequencing on the Illumina HiSeq4000 sequencer. RNA-sequencing data of wildtype and Esr2-null oocytes were analyzed, and differentially expressed genes were identified using the CLC Genomics Workbench. Whole oocyte transcriptome data of wildtype and Esr2-null oocytes were compared to identify the differentially expressed genes. Raw data are deposited to the NCBI Sequence Read Archive (SRA) and analyzed data are presented in this data article. These datasets can be utilized to identify the gonadotropin-induced genes in oocytes that are ESR2-regulated and important to oocyte maturation.
RESUMEN
The liver helps maintain energy homeostasis by synthesizing and storing glucose and lipids. Gonadal steroids, particularly estrogens, play an important role in regulating metabolism. As estrogens are considered female hormones, metabolic disorders related to the disruption of estrogen signaling have mostly been studied in females. Estrogen receptor alpha (ESR1) is the predominant receptor in both the male and female liver, and it mediates the hepatic response to estrogens. Loss of ESR1 increases weight gain and obesity in female rats, while reducing the normal growth in males. Although Esr1-/- male rats have a reduced body weight, they exhibit increased adipose deposition and impaired glucose tolerance. We further investigated whether these metabolic disorders in Esr1-/- male rats were linked with the loss of transcriptional regulation by ESR1 in the liver. To identify the ESR-regulated genes, RNA-sequencing was performed on liver mRNAs from wildtype and Esr1-/- male rats. Based on an absolute fold change of ≥2 with a p-value ≤ 0.05, a total of 706 differentially expressed genes were identified in the Esr1-/- male liver: 478 downregulated, and 228 upregulated. Pathway analyses demonstrate that the differentially expressed genes include transcriptional regulators (Cry1, Nr1d1, Nr0b2), transporters (Slc1a2), and regulators of biosynthesis (Cyp7b1, Cyp8b1), and hormone metabolism (Hsd17b2, Sult1e1). Many of these genes are also integral parts of the lipid and carbohydrate metabolism pathways in the liver. Interestingly, certain critical regulators of the metabolic pathways displayed a sexual dimorphism in expression, which may explain the divergent weight gain in Esr1-/- male and female rats despite common metabolic dysfunctions.