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The ARRIVE Guidelines. (DOC 37Â kb)

Intron 1 of Ccnb3, 578 bp other sequence and 5' splice site of 31 bp and 92 bp intron 1 retention splice mutants. (PDF 310Â kb)

Supplemental_figures_and_tables for The Notch Family Transcription Factor, RBPJκ, Modulates Glucose Transporter and Ovarian Steroid Hormone Receptor Expression During Decidualization by Michael R Strug, Ren-Wei Su, Tae Hoon Kim, Jae-Wook Jeong, and Asgerally Fazleabas in Reproductive Sciences

The assembly of the blood–testis barrier (BTB) during postnatal development is crucial to support meiosis. However, the role of germ cells in BTB assembly remains unclear. Herein, Kit(W)/Kit(WV) mice were used as a study model. These mice were infertile, failing to establish a functional BTB to support meiosis due to c-Kit mutation. Transplantation of undifferentiated spermatogonia derived from normal mice into the testis of Kit(W)/Kit(WV) mice triggered functional BTB assembly, displaying cyclic remodeling during the epithelial cycle. Also, transplanted germ cells were capable of inducing Leydig cell testosterone production, which could enhance the expression of integral membrane protein claudin 3 in Sertoli cells. Early spermatocytes were shown to play a vital role in directing BTB assembly by expressing claudin 3, which likely created a transient adhesion structure to mediate BTB and cytoskeleton assembly in adjacent Sertoli cells. In summary, the positive modulation of germ cells on somatic cell function provides useful information regarding somatic–germ cell interactions.—Li, X.-Y., Zhang, Y., Wang, X.-X., Jin, C., Wang, Y.-Q., Sun, T.-C., Li, J., Tang, J.-X., Batool, A., Deng, S.-L., Chen, S.-R., Cheng, C. Y., Liu, Y.-X. Regulation of blood–testis barrier assembly in vivo by germ cells.

Spermatozoa maturation, a process required for spermatozoa to acquire progressive motility and the ability to fertilize ova, primarily occurs in the caput and corpus of the epididymis. Despite considerable efforts, the factor(s) promoting epididymal sperm maturation remains unclear. Recently, WNT signaling has been implicated in epididymal sperm maturation. To further investigate WNT signaling function in epididymal sperm maturation, we generated Wntless conditional knockout mice (Wls cKO), Wls flox/flox ; Lcn5-Cre. In these mice, WNTLESS (WLS), a conserved membrane protein required for all WNT protein secretion, was specifically disrupted in the principal cells of the caput epididymidis. Immunoblot analysis showed that WLS was significantly reduced in the caput epididymidis of Wls cKO mice. In the caput epididymidis of Wls cKO mice, WNT 10A and WNT 2b, which are typically secreted by the principal cells of the caput epididymis, were not secreted. Interestingly, sperm motility analysis showed that the WLS deficiency in the caput epididymidis had no effect on sperm motility. Moreover, fertility tests showed that Wls cKO male mice had normal fertility. These results indicate that the disruption of WLS in principal cells of the caput epididymidis inhibits WNT protein secretion but has no effect on sperm motility and male fertility, suggesting that WNT signaling in the caput epididymidis may be dispensable for epididymal sperm maturation in mice.

Mammalian oocyte chromosomes undergo 2 meiotic divisions to generate haploid gametes. The frequency of chromosome segregation errors during meiosis I increase with age. However, little attention has been paid to the question of how aging affects sister chromatid segregation during oocyte meiosis II. More importantly, how aneuploid metaphase II (MII) oocytes from aged mice evade the spindle assembly checkpoint (SAC) mechanism to complete later meiosis II to form aneuploid embryos remains unknown. Here, we report that MII oocytes from naturally aged mice exhibited substantial errors in chromosome arrangement and configuration compared with young MII oocytes. Interestingly, these errors in aged oocytes had no impact on anaphase II onset and completion as well as 2-cell formation after parthenogenetic activation. Further study found that merotelic kinetochore attachment occurred more frequently and could stabilize the kinetochore-microtubule interaction to ensure SAC inactivation and anaphase II onset in aged MII oocytes. This orientation could persist largely during anaphase II in aged oocytes, leading to severe chromosome lagging and trailing as well as delay of anaphase II completion. Therefore, merotelic kinetochore attachment in oocyte meiosis II exacerbates age-related genetic instability and is a key source of age-dependent embryo aneuploidy and dysplasia.

Spermatogenesis is crucial for male fertility and is therefore tightly controlled by a variety of epigenetic regulators. However, the function of enhancer of zeste homolog 2 (EZH2) in spermatogenesis and the molecular mechanisms underlying its activity remain poorly defined. Here, we demonstrate that deleting EZH2 promoted spermatogonial differentiation and apoptosis. EZH2 is expressed in spermatogonia, spermatocytes and round and elongated spermatids from stage 9 to 11 but not in leptotene and zygotene spermatocytes. Knocking downEzh2 in vitrousing a lentivirus impaired self-renewal in spermatogonial stem cells (SSCs), and the conditional knockout ofEzh2in spermatogonial progenitors promoted precocious spermatogonial differentiation. EZH2 functions to balance self-renewal and differentiation in spermatogonia by suppressing NEUROG3 and KIT via a direct interaction that is independent of its histone methyltransferase activity. Moreover, deletingEzh2enhanced the activation of CASP3 in spermatids, resulting in reduced spermatozoa production. Collectively, these data demonstrate that EZH2 plays a nonclassical role in the regulation of spermatogonial differentiation and apoptosis in murine spermatogenesis.

Understanding the cocatalyst/semiconductor interaction is of key importance for the design and synthesis of next generation photocatalytic materials for efficient hydrogen production and environmental cleanup applications. Here we investigate preformed Pd nanoparticles (NPs) supported on a series of anatase TiO2 having well-controlled but varying degrees of crystallinity and crystallite size, and explore their photocatalytic performance for H2 production and phenol decomposition. While tuning the anatase crystallite size significantly influences the photocatalytic performance, varying the TiO2 crystallinity shows a negligible effect. Interestingly, the optimum quantum efficiency (∼78%) for H2 evolution is achieved with anatase having medium crystallite size (∼16 nm), whereas for phenol decomposition, a promotional effect is only observed for anatase with larger crystallite sizes (>20 nm). Surface radical species and radical densities study reveal that the photogenerated charge carriers have been trapped at different sites depending on the crystallite size of anatase. While the excited electrons are only trapped in bulk lattice sites in small anatase (

Spermatogenesis is a complex process involving the regulation of multiple cell types. As the only somatic cell type in the seminiferous tubules, Sertoli cells are essential for spermatogenesis throughout the spermatogenic cycle. The Wilms tumor gene,Wt1, is specifically expressed in the Sertoli cells of the mouse testes. In this study, we demonstrated thatWt1is required for germ cell differentiation in the developing mouse testes. At 10 dayspost partum,Wt1-deficient testes exhibited clear meiotic arrest and undifferentiated spermatogonia accumulation in the seminiferous tubules. In addition, the expression of claudin11, a marker and indispensable component of Sertoli cell integrity, was impaired inWt1−/flox;Cre-ERTMtestes. This observation was confirmed inin vitrotestis cultures. However, the basal membrane of the seminiferous tubules inWt1-deficient testes was not affected. Based on these findings, we propose that Sertoli cells' status is affected inWt1-deficient mice, resulting in spermatogenesis failure.

There are thought to be two forms of inhibition of return (IOR) depending on whether the oculomotor system is activated or suppressed. When saccades are allowed, output-based IOR is generated, whereas input-based IOR arises when saccades are prohibited. In a series of 4 experiments, we mixed or blocked compatible and incompatible trials with saccadic or manual responses to investigate whether cueing effects would follow the same pattern as those observed with more traditional peripheral onsets and central arrows. In all experiments, an uninformative cue was displayed, followed by a cue-back stimulus that was either red or green, indicating whether a compatible or incompatible response was required. The results showed that IOR was indeed observed for compatible responses in all tasks, whereas IOR was eliminated for incompatible trials-but only with saccadic responses. These findings indicate that the dissociation between input- and output-based forms of IOR depends on more than just oculomotor activation, providing further support for the existence of an inhibitory cueing effect that is distinct to the manual response modality.

To study the relationship between the molecular main chain structure and the properties of amine based tetra-functional epoxy resins, especially for the thermal stabilities and the mechanical properties, N,N,N',N'-Tetraglycidyl-2,2-Bis[4-(4-aminophenoxy)phenyl]propane (TGBAPP) and N,N,N',N'-Tetraglycidyl-4,4'-diamino diphenyl ether (TGDDE) were cured with Methyl nadic anhydride (MNA). The thermal behavior of the cured epoxy resins were studied with the thermo-gravimetric analysis (TGA), and the glass transition temperature (Tg) were determined with the Dynamic Mechanical Analysis (DMA). Additionally, the mechanical properties of them were tested. The results indicated that the cured epoxy resin based on TGBAPP had better thermal stabilities and toughness.

Increases in the aneuploidy rate caused by the deterioration of cohesion with increasing maternal age have been well documented. However, the molecular mechanism for the loss of cohesion in aged oocytes remains unknown. In this study, we found that intracellular pH (pHi) was elevated in aged oocytes, which might disturb the structure of the cohesin ring to induce aneuploidy. We observed for the first time that full-grown germinal vesicle (GV) oocytes displayed an increase in pHi with advancing age in CD1 mice. Furthermore, during the in vitro oocyte maturation process, the pHi was maintained at a high level, up to ∼7.6, in 12-month-old mice. Normal pHi is necessary to maintain protein localization and function. Thus, we put forward a hypothesis that the elevated oocyte pHi might be related to the loss of cohesion and the increased aneuploidy in aged mice. Through the in vitro alkalinization treatment of young oocytes, we observed that the increased pHi caused an increase in the aneuploidy rate and the sister inter-kinetochore (iKT) distance associated with the strength of cohesion and caused a decline in the cohesin subunit SMC3 protein level. Young oocytes with elevated pHi exhibited substantially the increase in chromosome misalignment.

Spermatogonial stem cells (SSCs) are a subpopulation of undifferentiated spermatogonia located in a niche at the base of the seminiferous epithelium delimited by Sertoli cells and peritubular myoid (PM) cells. SSCs self-renew or differentiate into spermatogonia that proliferate to give rise to spermatocytes and maintain spermatogenesis. Glial cell line-derived neurotrophic factor (GDNF) is essential for this process. Sertoli cells produce GDNF and other growth factors and are commonly thought to be responsible for regulating SSC development, but limited attention has been paid to the role of PM cells in this process. A conditional knockout (cKO) of the androgen receptor gene in PM cells resulted in male infertility. We found that testosterone (T) induces GDNF expression in mouse PM cells in vitro and neonatal spermatogonia (including SSCs) co-cultured with T-treated PM cells were able to colonize testes of germ cell-depleted mice after transplantation. This strongly suggested that T-regulated production of GDNF by PM cells is required for spermatogonial development, but PM cells might produce other factors in vitro that are responsible. In this study, we tested the hypothesis that production of GDNF by PM cells is essential for spermatogonial development by generating mice with a cKO of the Gdnf gene in PM cells. The cKO males sired up to two litters but became infertile due to collapse of spermatogenesis and loss of undifferentiated spermatogonia. These studies show for the first time, to our knowledge, that the production of GDNF by PM cells is essential for undifferentiated spermatogonial cell development in vivo.

Mammalian mediator (MED) is a multi-protein coactivator that has been identified by several research groups. The involvement of the MED complex subunit 19 (MED 19) in the metastasis of lung adenocarcinoma cell line (H1299), which expresses the MED 19 subunit, was here investigated. When MED 19 expression was decreased by RNA interference H1299 cells demonstrated reduced clone formation, arrest in the S phase of the cell cycle, and lowered metastatic capacity. Thus, MED 19 appears to play important roles in the biological behavior of non-small cell lung carcinoma cells. These findings may be important for the development of novel lung carcinoma treatments.

The ablation properties of C/C-ZrC-SiC composites prepared by precursor infiltration and pyrolysis processes were studied by the H2-O2 ablation method. The results indicate that the C/C composite of 1.28 g•cm-3 was densified to C/C-ZrC-SiC composite of 1.42 g•cm-3 after infiltration and pyrolysis once, the density increased 10.94%. The linear ablation rate and mass ablation rate of the C/C-ZrC-SiC composites (ρ=1.42g•cm-3) were 7.22μm•s-1and 3.88mg•s-1, which decreased 79.31% and 65.46% comparing with C/C composite of 1.28 g•cm-3 respectively. The introduction of ZrC-SiC into the matrix greatly enhanced the anti-oxidative and ablation property of the C/C composite. The ablation mechanisms of the C/C-ZrC-SiC composites were supposed to be the synergistic effects of thermo-chemistry ablation (oxidation and sublimation) and mechanical erosion.

The ablation properties of C/C-ZrC-SiC composites prepared by precursor infiltration and pyrolysis processes were studied by the H2-O2 ablation method. The results indicate that the ablation rate of C/C-ZrC-SiC composites decreases with increasing density of the composites. The linear ablation rate and mass ablation rate of the C/C-ZrC-SiC composites (ρ=1.69g·cm-3) were 0.00211mm·s-1 and 0.5179mg·s-1, only 20.6% and 31.6% of those of C/C composites (ρ=1.78g·cm-3) respectively. The introduction of ZrC-SiC into the matrix greatly enhanced the anti-oxidative and ablation property of the C/C composite. The ablation mechanisms of the C/C-ZrC-SiC composites were supposed to be the synergistic effects of thermo-chemistry ablation (oxidation and sublimation) and mechanical erosion.

This study focuses on the conditions of preparation of C/C-SiC composites through polymer infiltration and pyrolysis method using hydrogen-containing polysiloxane as the precursor and divinylbenzene as cross-linking agent. The densities of different C/C composites (ρ=1.02, 1.21, 1.60, 1.78g·cm-3) increased to 1.24, 1.36, 1.69, 1.84g·cm-3respectively after infiltration and pyrolysis four times. The appropriate ratio of hydrogen-containing polysiloxane and divinylbenzene was 1/0.5. The appropriate cross-linking temperature was 250°C. In pyrolysis process, the appropriate increasing rate of pyrolysis temperature should be 50°C/h in the range of 300-800°C. SiC was formed when pyrolysis temperature reached 1550°C.The linear ablation rate and mass ablation rate of C/C-SiC composites were much lower than those of C/C composites.

SMAD4 is the central component of canonical signaling in the transforming growth factor beta (TGFβ) superfamily. Loss of Smad4 in Sertoli cells affects the expansion of the fetal testis cords, whereas selective deletion of Smad4 in Leydig cells alone does not appreciably alter fetal or adult testis development. Loss of Smad4 in Sertoli and Leydig cells, on the other hand, leads to testicular dysgenesis, and tumor formation in mice. Within the murine testes, Smad4 is also expressed in germ cells of the seminiferous tubules. We therefore, crossed Ngn3-Cre or Stra8-Cre transgenic mice with Smad4-flox mice to generate conditional knockout animals in which Smad4 was specifically deleted in postnatal germ cells to further uncover cell type-specific requirement of Smad4. Unexpectedly, these germ-cell-knockout mice were fertile and did not exhibit any detectable abnormalities in spermatogenesis, indicating that Smad4 is not required for the production of sperm; instead, these data indicate a cell type-specific requirement of Smad4 primarily during testis development. Mol. Reprod. Dev. 83: 615-623, 2016. © 2016 Wiley Periodicals, Inc.

Testis cords, embryonic precursors of the seminiferous tubules, are fundamental for testis structure and function. Delay or disruption of testis cord formation could result in gonadal dysgenesis. Although mechanisms regulating testis cord formation during sex determination have been well-studied, the genes and signaling pathways involving in testis cord maintenance after the cords have formed are not well characterized. It is now clear that the maintenance of cord structure is an active process. In this review, we summarize the recent findings regarding the regulation of testis cord integrity by a series of Sertoli cell transcription factors, including the WT1-SOX8/SOX9-beta-CATENIN-DHH network, GPR56, STIM1, and NR0B1 (also known as DAX1). In particularly, we emphasize the underappreciated role of peritubular myoid cells in testis cord maintenance and their cooperation with Sertoli cells. The regulation of the size, shape, and number of testis cords by Sertoli cell proliferation (e.g., SMAD4, GATA4, and TGF-beta signaling), Leydig cell products (e.g., ACTIVIN A), vascular development (a lesson learned from PDGF signaling), and available gonad space (as observed in Ift144 mutant mice) is also addressed. Further efforts and new genetic models are needed to unveil the gene networks and underlying mechanisms regulating testis cord integrity and morphology after sex determination.

Arts programmes are provided as part of the psychosocial rehabilitation for consumers with a mental illness. Consumers have reported that involvement in the arts has facilitated their recovery. This qualitative study was conducted to explore the ways in which involvement in an Australian community arts programme contributed to the recovery process. It also aimed to gain an understanding of intrinsic recovery, which focuses on personal satisfaction and meaning. Eight consumers with a mental illness who were members of a community-based arts programme participated in in-depth semi-structured interviews. The themes that were analysed thematically from the interviews included art as a medium of expression and self-discovery, and changes in internal conditions (spirituality, empowerment and self-validation). The findings revealed the usefulness of arts programmes in providing the external environment that facilitates internal recovery.