Development & reproduction最新文献

Equine chorionic gonadotropin (eCG), produced by the endometrial cups of the placenta after the first trimester, is a specific glycoprotein that displays dual luteinizing hormone (LH)-like and follicle-stimulating hormone (FSH)-like effects in non-equid species. However, in equidaes, eCG exhibits only LH-like activity. To identify the specific biological functions of glycosylated sites in eCG, we constructed the following site mutants of N- and O-linked glycosylation: eCGβ/αΔ56, substitution of α-subunit⁵⁶ N-linked glycosylation site; eCGβ-D/α, deletion of the O-linked glycosylation sites at the β-subunit, and eCGβ-D/αΔ56, double mutant. We produced recombinant eCG (rec-eCG) proteins in Chinese hamster ovary suspension (CHO-S) cells. We examined the biological activity of rec-eCG proteins in CHO-K1 cells expressing the eLH/CG receptor and found that signal transduction activities of deglycosylated mutants remarkably decreased. The EC₅₀ levels of eCGβ/αΔ56, eCGβ-D/α, and eCGβ-D/αΔ56 mutants decreased by 2.1-, 5.6-, and 3.4-fold, respectively, compared to that of wild-type eCG. The Rmax values of the mutants were 56%-80% those of wild-type eCG (141.9 nmol/10⁴ cells). Our results indicate that the biological activity of eCG is greatly affected by the removal of N- and O-linked glycosylation sites in cells expressing eLH/CGR. These results provide important information on rec-eCG in the regulation of specific glycosylation sites and improve our understanding of the specific biological activity of rec-eCG glycosylation sites in equidaes.

The purpose of this paper is to critique the links between science and scientific researchers, politics and capital surrounding vaccines and vaccination in a pandemic era. It also introduces standards for adapting ethical guidelines for research under public health emergencies to specific circumstances and contexts. It also introduces ethical standards to be applied to scientific research. that is, scientific relevance, social value, cooperative partnerships, reasonable risk-benefit costs, fair and voluntary participation, independent review, and equal moral respect for participants and affected communities. It also outlines the COVID-19 (coronavirus disease 2019) pandemic-consent and other research procedure modifications proposed by Oregon Health and Science University (OHSU). We are on the lookout for powerful capital and hegemonic groups inseparable from politics and important decision makers in the pandemic era - that is, scientists, supporters, and scientific civic groups. It underscores the need for an independent and formal scientific advisory body with the right balance between science and politics.

The incidence of infertility among individuals of reproductive age has been growing due to genetic and environmental factors, and considerable research efforts are focused on solving this issue. Ovarian development is an overly complex process in the body, involving the interaction between primordial germ cells and gonad somatic cells. However, follicles located in the center of the in vitro ovary are poorly formed owing to ovarian complexity, nutrient deficiency, and signaling deficiency. In the present study, we optimized methods for dissociating gonads and culture conditions for the in vitro generation of miniaturized ovaries. The gonads from embryos were dissociated into cell masses and cultured on a Transwell-COL membrane for 3-5 weeks. Approximately 12 follicles were present per in vitro ovary. We observed that miniaturized ovaries successfully matured to MII oocytes in vitro from 150 to 100 µm gonad masses. This method will be useful for investigating follicle development and oocyte production.

In previous reports, bisphenol A (BPA) exposure affects reproductive function in Drosophila melanogaster females. To test the maternal effect of BPA exposure on fly reproductive function, F0 mothers were exposed to 0, 0.1, 1, and 10 mg/L of BPA and the fecundity in F1 and F2 generations were checked. In this experiment, 1 and 10 mg/L BPA significantly decreased the fecundity of F1 females. Moreover, 0.1 and 1 mg/L BPA substantially reduced egg production in the F2 generation. These results suggested that maternal exposure to BPA at enviromentally relavant concnetrations reduces reproductive function in Drosophila melanogaster females and that this effect is transgenerational.

To determine whether the reproductive processes of sea bass, Lateolabrax japonicus, proceed normally after transportation from an outdoor net-cage into indoor tanks, we examined changes in the gonadosomatic index (GSI), histological gonadal tissue, and plasma levels of sex hormones (testosterone and estradiol-17ß) during their annual reproductive cycle. We also measured maturation and spawning across two sea water salinity levels (full and low salinity). Fecundity was estimated by the relationship between egg number and body size in female sea bass. Monthly changes in the GSI, histological gonadal tissues, and oocyte size showed both male and female sea bass reach final maturation in January and February, respectively, indicating that the spermiation of males occurs earlier than the spawning of females. The histological results indicated that the sea bass is a multiple spawner, similar to many marine teleosts, exhibiting group-synchronous oocyte development. Female maturation and spawning were enhanced in lower salinity seawater (29.6-31.0 psu) compared to that of normal salinity (34.5-35.1 psu). These results confirm that sea bass reproduction can occur successfully in captivity and imply that fertilized eggs can be collected from February to March. Additionally, our results show that lower salinity enhances oocyte maturation and spawning of female sea bass.

We report on the gonadal structure abnormality and intersexuality found from a survey regarding the reproductive potential of the rock bream, Oplegnathus fasciatus, along the southern coast of Korea. In total, 448 samples were collected for histological analysis (total length 27.5±6.8 cm; total weight 522.9±385.6 g). The sex ratio (F:M) was 1:0.46 (n=307:141), with 68.5% being females. The frequency of oocyte atresia and ovarian neoplasm in the females was 85.3% and 21.5%, respectively, and the frequency of testicular lobule and cyst deformation and testicular neoplasm in the males was 73.1% and 37.6%, respectively. The scale formation in the ovary and testis was approximately 2.0% for both sexes. The intersexuality was 42.2%, with the intersexuality among females being 49.5%, which was significantly higher than that of males (26.2%). Such findings indicate a negative impact on the reproductive output of the rock bream along the southern coast of Korea; however, a more detailed study is required to ascertain accurate causes.

Reproductive cycle of the blue-striped angelfish, Chaetodontoplus septentrionalis were histologically investigated. Fish were monthly collected in the coastal waters of Munseom, Seogwipo, Jeju-Island from February to December 2018. The gonadosomatic index (GSI) increased from May and maintained high values in August. The reproductive cycle of female fish can be classified by the characteristics observed during gonadal development as followed: growing stage (November to June), early mature stage (May to June), mature and spawning stage (June to September), and degenerative and recovery stage (September to December). In the male, testicular development period was similar to that of ovarian development period, but mature and spawning period was one month longer from June to October. Fecundity of mature female ranged from 4,601 to 22,840 and was correlated positively with total length and body weight. The histological analysis of gonadal development indicated that the C. septentrionalis was summer-spawning type and is considered a multiple spawner during spawning season.

In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.

Cutaneous melanoma is a fatal disease for patients with distant metastasis. Metformin is the most widely used anti-diabetic drug, and proved to suppress cell proliferation and metastasis in diverse cancers including melanoma. We previously reported that MEK inhibitor trametinib increases the expression of epithelial-mesenchymal transition (EMT) regulators and melanoma cell motility, which are suppressed by addition of metformin in A375 melanoma cells. To confirm our findings further, we first evaluated the effect of metformin in combination with another MEK inhibitor binimetinib on cell viability in G361 melanoma cells. We then investigated whether binimetinib affects the expression of EMT regulators and cell motility. We finally monitored the effect of metformin on binimetinib-induced cell migration. Cell viability assay showed that combination index (CI) value at ED₅₀ is 0.80, suggesting synergy for the combination of metformin with binimetinib. Our results also revealed that binimetinib increased the expression of EMT regulators such as integrin αV, fibronectin and slug, which correlate well with the enhanced cell migration in wound healing assay. Metformin, on the contrary, suppressed the expression of sparc, integrin αV, fibronectin and N-cadherin with the reduced cell motility. The combination treatment showed that metformin counteracts the binimetinib-induced increase of cell motility. Overall, these results suggest that metformin with binimetinib might be useful as a potential therapeutic adjuvant against cell survival and metastatic activity in melanoma patients.