Mutations in TUBB8 and Human Oocyte Meiotic Arrest

BACKGROUND Human reproduction depends on the fusion of a mature oocyte with a sperm cell to form a fertilized egg. The genetic events that lead to the arrest of human oocyte maturation are unknown. METHODS We sequenced the exomes of five members of a four-generation family, three of whom had infertility due to oocyte meiosis I arrest. We performed Sanger sequencing of a candidate gene, TUBB8, in DNA samples from these members, additional family members, and members of 23 other affected families. The expression of TUBB8 and all other β-tubulin isotypes was assessed in human oocytes, early embryos, sperm cells, and several somatic tissues by means of a quantitative reverse- transcriptase–polymerase-chain-reaction assay. We evaluated the effect of the TUBB8 mutations on the assembly of the heterodimer consisting of one α-tubulin polypeptide and one β-tubulin polypeptide (α/β-tubulin heterodimer) in vitro, on microtubule architecture in HeLa cells, on microtubule dynamics in yeast cells, and on spindle assembly in mouse and human oocytes. RESULTS We identified seven mutations in the primate-specific gene TUBB8 that were responsible for oocyte meiosis I arrest in 7 of the 24 families. TUBB8 expression is unique to oocytes and the early embryo, in which this gene accounts for almost all the expressed β-tubulin. The mutations affect chaperone-dependent folding and assembly of the α/β-tubulin heterodimer, disrupt microtubule behavior on expression in cultured cells, alter microtubule dynamics in vivo, and cause catastrophic spindle-assembly defects and maturation arrest on expression in mouse and human oocytes. CONCLUSIONS TUBB8 mutations have dominant-negative effects that disrupt microtubule behavior and oocyte meiotic spindle assembly and maturation, causing female infertility. (Funded by the National Basic Research Program of China and others.

Global urban environmental change drives adaptation in white clover

Urbanization transforms environments in ways that alter biological evolution. We examined whether urban environmental change drives parallel evolution by sampling 110,019 white clover plants from 6169 populations in 160 cities globally. Plants were assayed for a Mendelian antiherbivore defense that also affects tolerance to abiotic stressors. Urban-rural gradients were associated with the evolution of clines in defense in 47% of cities throughout the world. Variation in the strength of clines was explained by environmental changes in drought stress and vegetation cover that varied among cities. Sequencing 2074 genomes from 26 cities revealed that the evolution of urban-rural clines was best explained by adaptive evolution, but the degree of parallel adaptation varied among cities. Our results demonstrate that urbanization leads to adaptation at a global scale

Climate Change and Its Attribution in Three Gorges Reservoir Area, China

Climate change in dam areas is one of the environmental problems associated with dams. However, the main factors and mechanisms that impact climate change in dam areas remain unclear. In this study, linear regression, the observed minus reanalyzed (OMR) method, and multi-source data are used to assess climate change in the Three Gorges Reservoir Area of China and investigate the main impact factors among the controversial factors (land cover change, environmental climate, and reservoir impoundment). Our results indicate that turning points of trend changes for annual fog days (FD), annual average temperature (T), and annual average relative humidity (RH) occurred at around 1996 during the period 1973–2013, and annual precipitation (PRE) suggested no obvious turning point. The change trends after 1996 were steeper than before 1996. These changes are mainly closely correlated with environmental climate. In particular, temperature was significantly correlated with environmental temperature (1979–2013: r = 0.799, p < 0.01), and their relationship was stronger after 1990 (r = 0.842, p < 0.01). Moreover, the turning point for FD, T, and RH also correlated with land use/cover change. In addition, reservoir impoundment showed an obvious humidification effect (OMR RH correlated with water area: r = 0.566, p < 0.01). Our findings support the view that climate change in dam areas is mainly affected by environmental climate changes

Downregulation of SPARC expression inhibits the invasion of human trophoblast cells in vitro.

Successful pregnancy depends on the precise regulation of extravilloustrophoblast (EVT) invasion into the uterine decidua. SPARC (secreted protein acidic and rich in cysteine) is a matricellular glycoprotein that plays critical roles in the pathologies associated with obesity and diabetes, as well as tumorigenesis. The objective of this study was to investigate the role of SPARC in the process of trophoblast invasion which shares many similarities with tumor cell invasion. By Western blot, higher expression of SPARC was observed in mouse brain, ovary and uterus compared to other mouse tissues. Immunohistochemistry analysis revealed a spatio-temporal expression of SPARC in mouse uterus in the periimplantation period. At the implantation site of d8 pregnancy, SPARC mainly accumulated in the secondary decidua zone (SDZ), trophoblast cells and blastocyst. The expression of SPARC was also detected in human placental villi and trophoblast cell lines. In a Matrigel invasion assay, we found SPARC-specific RNA interference significantly reduced the invasion of human extravilloustrophoblast HTR8/SVneo cells. Microarray analysis revealed that SPARC depletion upregulated the expression of interleukin 11 (IL11), KISS1, insulin-like growth factor binding protein 4 (IGFBP4), collagen type I alpha 1 (COLIA1), matrix metallopeptidase 9 (MMP9), and downregulated the expression of the alpha polypeptide of chorionic gonadotropin (CGA), MMP1, gap junction protein alpha 1 (GJA1), et al. The gene array result was further validated by qRT-PCR and Western blot. The present data indicate that SPARC may play an important role in the regulation of normal placentation by promoting the invasion of trophoblast cells into the uterine decidua

Ecosystem services of lake-wetlands exhibit significant spatiotemporal heterogeneity and scale effects in a multi-lake megacity

Climate change and anthropogenic disturbances are altering global lake-wetlands (LWs) and their ecosystem services (ESs), posing a threat to sustainable development. Understanding the spatiotemporal heterogeneity and scale effects of LWESs has risen to be a critical concern in the sustainable management of wetlands. Although the LWESs of large lakes or at the macro-region are well documented, their complexity in urbanizing areas remains to be further addressed. Here we aim to reveal the spatiotemporal heterogeneity and scale effects of LWESs under the coinciding effects of wetland conservation and urbanization through a study in Wuhan, a multi-lake city of central China. The LWESs and their spatiotemporal variations from 1999 to 2019 were characterized by nine indicators based on time-series remote-sensing images and statistical data. A V-shaped trend was observed in the area of the LWs over time, which decreasing by 8.12% in 1999–2010 while recovering by 9.57% in 2010–2019. The LW loss in the main urban area (MA) was mainly encroached by construction lands, while the restoration of croplands to LWs in the rural area (RA) completely offsets the loss of LWs. We found that the change rates of most LWESs were more severely degraded along a rural–urban gradient, except for cultural services. Specifically, the water supply (-57.39%), flood regulation (-34.19%), climate regulation (-31.36%), and biodiversity conservation (-62.13%) services were the most severely damaged in LWs in the MA. However, the carbon sink services of LWs in the RA suffer the most severe degradation. Not only with spatiotemporal heterogeneity but LWESs also showed obvious scale effects. The larger the LWs, the higher the ES capacity tends to be. Smaller LWs were heavily encroached, with the worst ES capacities. These results highlight that LW conservation should no longer be homogeneous, but should combine indicators of scale effects and spatial heterogeneity. Due to land resource constraints, those close to urban center should improve regulating and supporting service capacities, while further away, improvements in area and LWESs can be pursued to balance the demand growth. Further, it is important to conserve existing and small LWs from over-encroachment

Data from: Improving conservation effectiveness of nature reserve for golden snub-nosed monkey, a niche-based approach

Reserve selections are often opportunistic rather than strategic and coordinated, and consequently many reserves are ineffective to achieve their intended goals of conservation. Here we assessed the conservation effectiveness of a reserve for the golden snub-nosed monkeys (Rhinopithecus roxellana) with a niche-based approach. We assessed habitat usage of the monkeys in Shennongjia Nature Reserve (SNR) and attributes of 14 environmental variables that could potentially affect the monkeys’ habitat use. Spatial distribution of potentially suitable habitat for the monkeys was then modeled with Maxent, a niche-based model, and conservation effectiveness of SNR was assessed by comparing the current boundary of the reserve with the spatial distribution of the modeled potential habitat and the current habitat area of the monkeys. Only 59% of the habitat area and 61% of the predicted potential habitat area were under the protection of SNR. To improve conservation effectiveness of SNR, we proposed that the current SNR be enlarged by 270 km2. The enlarged reserve would encompass 100% of the existing habitat area plus 89% of the predicted potential habitat area. Using the niche-based approach, we were able to integrate habitat usage data of the target species with that of remote sensing to identify areas potentially suitable as habitat for the species. This information not only can be used for improving conservation effectiveness of existing reserves but also for the effective planning and designing of new reserves

The ovarian estrogen synthesis function was impaired in Y123F mouse and partly restored by exogenous FSH supplement

Abstract Background LepR tyrosine site mutation mice (Y123F) exhibit decreased serum E2 levels, immature reproductive organs, infertility as well as metabolic abnormalities. Although the actions of leptin and lepR in the control of reproductive function are thought to be exerted mainly via the hypothalamic-pituitary-gonadal axis, relatively less is known regarding their local effects on the peripheral ovary, especially on steroid hormone synthesis. Meanwhile, whether the decreased fertility of Y123F mouse could be restored by gonadotropin has not been clear yet. Methods The serum levels of E2, P4, FSH, LH, T and leptin of Y123F and WT mice at the age of 12 weeks were measured by enzyme-linked immunosorbent assay. Immunohistochemistry was used to compare the distribution of hormone synthases (STAR, CYP11A1, CYP19A1, HSD17B7) and FSHR in adult mouse ovaries of two genotypes. Western blot and real-time PCR were used to detect the expression levels of four ovarian hormone synthases and JAK2-STAT3 / STAT5 signaling pathway in 4 and 12 weeks old mice, as well as the effects of exogenous hFSH stimulation on hormone synthases and FSHR. Results Compared with WT mice, the serum levels of FSH, LH and E2 in 12-week-old Y123F mice were significantly decreased; T and leptin levels were significantly increased; but there was no significant difference of serum P4 levels. STAR, CYP11A1, HSD17B7 expression levels and the phosphorylation levels of JAK2 and STAT3 were significantly decreased in adult Y123F mice, while the expression of CYP19A1 and phospho-STAT5 were significantly increased. No significant differences were found between 4-week-old Y123F and WT mice. After exogenous hFSH stimulation, E2 levels and expression of CYP19A1 and HSD17B7 were significantly higher than that in the non-stimulated state, but significant differences still existed between Y123F and WT genotype mice under the same condition. Conclusions Abnormal sex hormone levels of Y123F mice were due to not only decreased gonadotropin levels in the central nervous system, but also ovarian hormone synthase abnormalities in the peripheral gonads. Both FSH signaling pathway and JAK2-STAT3/STAT5 signaling pathway were involved in regulation of ovarian hormone synthases expression. Exogenous FSH just partly improved the blood E2 levels and ovarian hormone synthase expression

Winter hibernation and UCHL1-p34cdc2 association in toad oocyte maturation competence.

Currently, it is believed that toad oocyte maturation is dependent on the physiological conditions of winter hibernation. Previous antibody-blocking experiments have demonstrated that toad ubiquitin carboxyl-terminal hydrolase L1 (tUCHL1) is necessary for germinal vesicle breakdown during toad oocyte maturation. In this paper, we first supply evidence that tUCHL1 is highly evolutionarily conserved. Then, we exclude protein availability and ubiquitin carboxyl-terminal hydrolase enzyme activity as factors in the response of oocytes to winter hibernation. In the context of MPF (maturation promoting factor) controlling oocyte maturation and to further understand the role of UCHL1 in oocyte maturation, we performed adsorption and co-immunoprecipitation experiments using toad oocyte protein extracts and determined that tUCHL1 is associated with MPF in toad oocytes. Recombinant tUCHL1 absorbed p34(cdc2), a component of MPF, in obviously larger quantities from mature oocytes than from immature oocytes, and p13(suc1) was isolated from tUCHL1 with a dependence on the ATP regeneration system, suggesting that still other functions may be involved in their association that require phosphorylation. In oocytes from hibernation-interrupted toads, the p34(cdc2) protein level was significantly lower than in oocytes from toads in artificial hibernation, providing an explanation for the different quantities isolated by recombinant tUCHL1 pull-down and, more importantly, identifying a mechanism involved in the toad oocyte's dependence on a low environmental temperature during winter hibernation. Therefore, in toads, tUCHL1 binds p34(cdc2) and plays a role in oocyte maturation. However, neither tUCHL1 nor cyclin B1 respond to low temperatures to facilitate oocyte maturation competence during winter hibernation

What Is the Relationship between Natural Protected Areas and Stakeholders? Based on Literature Analysis from 2000–2021

The establishment of natural protected areas (NPAs) is an effective means to deal with the degradation of ecosystems caused by climate change and human activities. The area and number of NPAs in the world have shown an obvious growth trend, and their development has ushered in a new bottleneck. More importantly, the management quality of NPAs should be improved, and the key to improving management quality lies in human beings, but the stakeholder groups involved in NPAs are often overlooked by policymakers. In this study, a quantitative review of the global scientific literature on NPAs stakeholders was conducted using a bibliometric approach. The research hotspots and trends, number, time, and countries were analyzed based on data from published articles. The stakeholder types and internal relationships in NPAs were summarized and mapped. The common problems of resources and community resident management among stakeholders were discussed. A total of 5584 research articles selected from the Web of Science core collection database were used as data sources and were visualized using VOSviewer and the Biblioshiny program in the R language. The results of the study help to reveal the mutual influence mechanism between stakeholders during the development of nature reserves and contribute to the sustainable development of global protected areas and human well-being

Caulis Sargentodoxae Prescription Plays a Therapeutic Role with Decreased Inflammatory Cytokines in Peritoneal Fluid in the Rat Endometriosis Model

Caulis Sargentodoxae prescription has been confirmed by the gynecological clinical observation to be effective in the treatment of endometriosis (EMs), and inflammatory cytokines were involved in EMs. In this paper, animal experiments were designed to explain anti-inflammatory mechanisms of Caulis Sargentodoxae prescription on endometriosis. The EMs model was established by autoplastic transplantation, and rats were randomly divided into seven groups: normal control group, model group, ovariectomized group, gestrinone (Western medicine) group, Caulis Sargentodoxae prescription (Chinese medicine) group, celecoxib (inhibitor) group, and combination (Chinese medicine + inhibitor) group. After oral administration for 21 days, the growth inhibitory rates of the ectopic endometria in treatment groups were evaluated, and the levels of inflammatory cytokines in the serum and peritoneal fluid were determined by ELISA, as well as the expression of prostaglandin endoperoxide synthase 2 (PTGS2) in the ectopic endometrial tissues was detected by real-time polymerase chain reaction, immunohistochemistry, and western blotting. The growth inhibitory rates of the ectopic endometria were significantly higher in the Caulis Sargentodoxae prescription group and gestrinone group, in comparison with the model group p<0.05. In the Caulis Sargentodoxae prescription group, the levels of inflammatory cytokines including IL-1, IL-2, IL-6, TNF-α, and PGE2 were all reduced in the serum and peritoneal fluid p<0.05. In addition, the specific expression of PTGS2 in the ectopic endometrial tissues significantly decreased in the Caulis Sargentodoxae prescription group and PTGS2 inhibitor celecoxib group both at mRNA and protein levels, but in the steroid hormone drug gestrinone group not at the mRNA level. So, Caulis Sargentodoxae prescription has a reliable therapeutic effect on the EMs by its comprehensive anti-inflammatory roles, possibly in a way different from gestrinone