LncRNA STAT3-AS regulates endometrial receptivity via the STAT3 signaling pathway.

Endometrial receptivity is critical for the successful establishment of pregnancy in ruminants. Interferon tau (IFNT) plays a key role in promoting embryo attachment by activating the Janus kinase/signal transducer and activator of transcription pathway, which induces the expression of a series of interferon-stimulated genes (ISGs). In our previous study, sequencing analysis of goat endometrial epithelial cells (gEECs) treated with 20 ng/mL IFNT revealed a differentially expressed long non-coding RNA located on the STAT3 antisense chain, which we designated STAT3-AS. The aim of this study was to investigate the role and mechanism of STAT3-AS in establishing endometrial receptivity in goats. The results showed that STAT3-AS was expressed in both the nucleus and cytoplasm of gEECs, and its expression increased significantly in the uterus on day 15 of pregnancy. STAT3-AS expression was upregulated in gEECs treated with IFNT alone or in combination with progesterone and estradiol. Knockdown of STAT3-AS using specific short interfering RNA significantly inhibited the expression of classical ISGs such as interferon-stimulated gene 15 and 2',5'-oligodenylate synthetase 2, as well as uterine endometrial receptivity-related genes including homeobox gene A11, integrin beta 3, and vascular endothelial growth factor. Moreover, gEEC proliferation and the STAT3 pathway were suppressed in the absence of STAT3-AS. However, pretreatment with the STAT3 activator RO8191 restored the effect of silencing STAT3-AS on endometrial receptivity. Overall, these results suggest that STAT3-AS is an important regulator of endometrial receptivity in goats and that it regulates endometrial receptivity through the STAT3 pathway.

USP18 promotes endometrial receptivity via the JAK/STAT1 and the ISGylation pathway.

Interferon-tau (IFNT), a pregnancy recognition signal in ruminants, promotes the establishment of endometrial receptivity by inducing the expression of interferon-stimulated genes (ISGs) via the Janus kinase/signal transducer and activator of transcription (JAK/STATs) signaling pathway. However, the precise mechanisms remain largely unknown. Ubiquitin-specific protease 18 (USP18) acts specifically on the ISGylation modification system to exert deubiquitination and participates in the regulation of the type I IFN signaling pathway. The purpose of this study was to determine the role and mechanism of USP18 on endometrial receptivity in goat. USP18 was mainly localized in the uterine luminal and glandular epithelium, and its expression levels were significantly increased from days 5-18 of early pregnancy. Progesterone (P4), estradiol (E2), and IFNT significantly stimulated USP18 expression in goat endometrial epithelial cells (gEECs) cultured in vitro. Meanwhile, the markers of endometrial receptivity HOXA11, ITGB1, ITGB3, and ITGB5 were significantly upregulated after USP18 overexpression in gEECs. However, USP18 interference significantly inhibited the expression of HOXA10, ITGB1, ITGB3, and ITGB5 in gEECs. In addition, both the phosphorylation levels of STAT1 and the expression of ISGylation-modified proteins were significantly increased after USP18 silencing in gEECs. Furthermore, pretreatment with the STAT1 inhibitor Fludara markedly restored the effect of USP18 interference in gEECs. In summary, USP18 may play an important role in promoting goat endometrial receptivity by regulating the JAK/STAT1 pathway and ISGylation.

Interferon-τ -induced ISG15-AS regulates endometrial receptivity during early goat pregnancy.

Endometrial receptivity is a critical process for the successful establishment of pregnancy in ruminants. However, the biological role of long non-coding RNAs (lncRNAs) in the development of endometrial receptivity is poorly understood. In this study, we performed RNA-seq analysis of immortalised goat endometrial epithelial cells (gEECs) treated with interferon-τ (IFNT). Transcriptome profiles showed that 8069 high-confidence putative lncRNAs, including 6498 intronic lncRNA transcripts, 1078 lincRNAs and 493 antisense lncRNAs were identified in gEECs with or without IFNT treatment. Functional clustering analysis was performed by using cis and trans lncRNAs prediction. GO and KEGG analyses revealed that differentially expressed lncRNAs may regulate tissue remodelling and immune responses. Subsequently, six of the 21 differentially expressed antisense lncRNAs were validated using qRT-PCR. Through functional screening and co-expression analysis of lncRNAs in gEECs, we identified that ISG15-AS was mainly expressed in the luminal and glandular epithelium on days 5 and 15 and was strongly upregulated on day 18 of pregnancy in vivo. Similarly, ISG15-AS was abundant in the nucleus and cytoplasm, and was significantly upregulated after treatment with IFNT in gEECs. In addition, ISG15 is an IFNT-responsive gene, that displayed an evident increase in vivo and in vitro. Moreover, sense ISG15 was significantly upregulated following ISG15-AS silencing. The key genes related to ISGylation and endometrial receptivity in gEECs dramatically increased after ISG15-AS inhibition. Collectively, our results indicate that a novel antisense lncRNA, ISG15-AS, may be important in regulating endometrial receptivity through ISGylation.

Hand2os1 Regulates the Secretion of Progesterone in Mice Corpus Luteum.

The corpus luteum plays a key role in pregnancy maintenance and estrous cycle regulation by secreting progesterone. Hand2os1 is an lncRNA located upstream of Hand2, with which a bidirectional promoter is shared and is involved in the regulation of cardiac development and embryo implantation in mice. The aim of this study was to investigate the expression and regulation of Hand2os1 in the ovaries. Here, we used RNAscope to detect differential expression of Hand2os1 in the ovaries of cycling and pregnant mice. Hand2os1 was specifically detected in luteal cells during the proestrus and estrus phases, showing its highest expression in the corpus luteum at estrus. Additionally, Hand2os1 was strongly expressed in the corpus luteum on day 4 of pregnancy, but the positive signal progressively disappeared after day 8, was detected again on day 18, and gradually decreased after delivery. Hand2os1 significantly promoted the synthesis of progesterone and the expression of StAR and Cyp11a1. The decreased progesterone levels caused by Hand2os1 interference were rescued by the overexpression of StAR. Our findings suggest that Hand2os1 may regulate the secretion of progesterone in the mouse corpus luteum by affecting the key rate-limiting enzyme StAR, which may have an impact on the maintenance of pregnancy.

High-resolution mapping of the global silicate weathering carbon sink and its long-term changes.

Climatic and non-climatic factors affect the chemical weathering of silicate rocks, which in turn affects the CO2 concentration in the atmosphere on a long-term scale. However, the coupling effects of these factors prevent us from clearly understanding of the global weathering carbon sink of silicate rocks. Here, using the improved first-order model with correlated factors and non-parametric methods, we produced spatiotemporal data sets (0.25° × 0.25°) of the global silicate weathering carbon-sink flux (SCSFα ) under different scenarios (SSPs) in present (1950-2014) and future (2015-2100) periods based on the Global River Chemistry Database and CMIP6 data sets. Then, we analyzed and identified the key regions in space where climatic and non-climatic factors affect the SCSFα . We found that the total SCSFα was 155.80 ± 90 Tg C yr-1 in present period, which was expected to increase by 18.90 ± 11 Tg C yr-1 (12.13%) by the end of this century. Although the SCSFα in more than half of the world was showing an upward trend, about 43% of the regions were still showing a clear downward trend, especially under the SSP2-4.5 scenario. Among the main factors related to this, the relative contribution rate of runoff to the global SCSFα was close to 1/3 (32.11%), and the main control regions of runoff and precipitation factors in space accounted for about 49% of the area. There was a significant negative partial correlation between leaf area index and silicate weathering carbon sink flux due to the difference between the vegetation types. We have emphasized quantitative analysis the sensitivity of SCSFα to critical factors on a spatial grid scale, which is valuable for understanding the role of silicate chemical weathering in the global carbon cycle.

Response of the weathering carbon sink in terrestrial rocks to climate variables and ecological restoration in China.

The weathering carbon sink (CS) of rocks has a sensitive response to different influencing factors, and it is important to accurately distinguish this response in the global carbon cycle. However, no quantitative analysis of the response mechanism has been performed. In this study, the CS of the 12 types of terrestrial rocks in China from 2000 to 2014 is estimated using the GEM-CO2 model. The relative contribution rates of climate change and ecological restoration to the CS are quantitatively evaluated using the Lindeman-Merenda-Gold model. Results showed that: (1) The CS of terrestrial rocks in China was 17.69 Tg C yr-1, and the CS flux (CSF) was 2.53 t C km-2 yr-1; mixed sedimentary rocks had the highest CS (6.89 Tg C yr-1), and carbonate rocks had the highest CSF (5.8 t C km-2 yr-1). (2) The average annual CSF slightly decreased at a rate of 5.4 kg C km-2 yr-1; the areas of the CSF that decreased in the south were the areas where water budget decreased significantly, and it was the areas with a reduced water budget and ecological deterioration in the north. (3) The relative contribution rates of water budget and precipitation reached 57% and 35%, respectively; the response of the CSF to temperature was evident in areas with low or decreasing temperatures, and the influence of fractional vegetation cover (FVC) on the CSF in low value area was evident. (4) Mixed sedimentary rocks and carbonate rocks displayed a more evident reduction trend in the CSF than other rocks. This research verified the applicability of the GEM-CO2 model in China and presented a scientific basis for quantitative assessment of the impact of climate change and ecological restoration on the CSF.