GOAL
To investigate the role of ricter / MTTORC2 in the formation of the anti-chirmer barrier (BTB), testicular development and spermatogenesis.
Methods AMH CRE positive mice are homozygous for RICTOR LOXP with a specific RICTOR sertoli cell removal have been obtained by AMH CRE cross-high mice with RICTOR LOXP mice. The histology of the reproduction members, seminiferous tubules and the epididymis of transgenic mice were observed with coloring. The germ cell cell subgroups in the seminiferous tubule were detected by flow cytometry with labeling of propidium iodide. Ki 67 and sepaase expression levels were detected with immunofluorescence assay and BTB-associated protein expression levels were detected with immunofluorescence and Western blots.
RESULTS Compared to control (AMH CREF, RICTORLOXP / LOXP or RICTORLOXP / -), SERTOLI specific cell deleting mice has decreased considerably the weight of testicular weight and the epididymium weight (p <0.05), increased by Significantly a significant level of diploid cells (p <0.01) and decreased haploid cells (p <0.01) but comparable tetraploid cells and similar levels of ki 67 and sepaase. The mice with Knockout Ricor also showed an aberrant location of the BTB associated proteins, which were dispersed over the entire seminiferous epithelium, but the expression levels of the protein remained stable.
Conclusions
The testicular sertoli cells are essential for maintaining the integrity and function of BTB and ensuring normal spermatogenesis in the mouse.
MPS1 Kinase dependent on the location of median kinase-dependent centromere Protection of coheche in mice meiosis of mice I.
A main characteristic of meiosis is the removal of step-by-step cohesin of cohesin, the protein complex holding sister chromatids together, first of all weapons in meiosis I and then the centromere region of Méiose II. The centromeric cohesia is protected by SGO2 of sepaase-mediated cleavage, in order to maintain sister chromatids together until they separate from meiosis II. The failures in the elimination of step-by-step cohesic have resulted in aneuploid gametes, thus preventing the generation of healthy embryos.
Here, we note that Kinase activities of Bub1 and MPS1 are needed for SGO2 location in the centromere region. The oocytes treated with the inhibitor of the MPS1 are defective in the protection of centromeric cohesic, while oocytes devoid of a Bub1 kinase activity, which can not be phosphorylate H2A to T121, are not disturbed in the protection of cohesin. As long as MPS1 is functional. The activities MPS1 and Bub1 Kinase respectively locate the SGO2 in the meiosis I preferably at the centromera and the pericentromer, indicating that SGO2 at the centromera is required for protection.in Meiosis I Centromeric Cohesin is protected by SGE2 from the mediation cleavage of Sepase ensuring that sister chromatics are maintained together until they separation in meiosis II.
Here, the authors show that Bub1 and MPS1 Kinase activities are required for SGO2 location in the Centromere area. Sepased, an enzyme that solves the cohesion of sister chromatids during the transition of metaphase-to-anaphase, plays a central role in chromosomal segregation and cell division. Sepased protein, encoded by additional spindle pole bodies such as 1 gene (ESPL1), is overexpressed in many human cancers, including chest, bones, brain and prostate. Sepase is oncogene and overexpression is sufficient to induce mammary tumors in mice. Acute or chronic overexpression of sepaase in the mute glands leads to aneuploidy and tumorigenesis and the inhibition of the enzymatic separation activity decreases the growth of the xenograms of the human chest tumor in the mouse.
[RICTOR / MTORC2 regulates the barrier of chronology and spermatogenesis in mice].
The related polo-like kinase made determining the distribution of bub1 in meiosis I.
In meiosis I, chromatid brother was arrested by the same pole (mono orientation), and central cohesion protected throughout the anafase. Shugoshin, which is litkromeres depends on the phosphorylation of Histone H2A by Butug Kinase, plays a central role in protecting cohesin meiotic rec8 from separation division. Meiotic Kinetochore Factors Other Keys, Meikin, can regulate cohesion protection, even though the underlying molecular mechanism is still difficult to understand. Here, we showed that MOA1 yeast fission (Meikin), which colleagues with CENP-C during Meiosis I, recruited PLO1 (polo-like kinase) to Kinetochores and Fosforilates SPC7 (KNL1) to accumulate BUB1. As a result, in contrast to the localization of the release of the transient Kinetochore from Mitosis Bub1, Meiother Bub1 survive in Kinetochores to Anafase I. Pond Bub1 Meiotic ensured the protection and protection of strong cohesion in the centromeres by working with Heterocromatin Swi6 protein, which binded and stabilized SGO1.
Furthermore, molecular genetic analysis shows hierarchical regulations of coconaceric cohesion protection by Meikin and Shugoshin which are important to establish special chromosome segregations of meiosis. We provide evidence that the BUB1 regulation specific meiosis is preserved in the mouse. Mamalia Nima-Like Kinase-1 (NEK1) is a multiple specific kinase which is very stated in mouse germ cells during the Iiosis prophase. Losing NEX1 induces cohesin retention on chromosomes in Meiotic prophase I.
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates, sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1, or separase, which initiates the final separation of sister chromatids.
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates, sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1, or separase, which initiates the final separation of sister chromatids.
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801
Description: A polyclonal antibody for detection of Separase phospho Ser801) from Human, Mouse. This Separase phospho Ser801) antibody is for WB , IHC-P, IF, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from human Separase around the phosphorylation site of S801
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates| sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1| or separase| which initiates the final separation of sister chromatids (Sun et al.| 2009
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates| sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1| or separase| which initiates the final separation of sister chromatids (Sun et al.| 2009
Description: Stable cohesion between sister chromatids before anaphase and their timely separation during anaphase are critical for chromosome inheritance. In vertebrates| sister chromatid cohesion is released in 2 steps via distinct mechanisms. The first step involves phosphorylation of STAG1 (MIM 604358) or STAG2 (MIM 300826) in the cohesin complex. The second step involves cleavage of the cohesin subunit SCC1 (RAD21; MIM 606462) by ESPL1| or separase| which initiates the final separation of sister chromatids (Sun et al.| 2009
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human ESPL1 / Separase (aa767-816). This antibody is tested and proven to work in the following applications:
Deposition on time and the elimination of cohesin is very important for accurate chromosome separation. Two processes regulate the removal of cohesin: non-proteolytic mechanism involving wapl, sororin, and PDS5B and direct cleavage with a separase. Here, we showed the role of NEK1 in the WAPL loading regulation during Meiotic Prophase I, through interactions between Nek1 and PDS5b.
![[RICTOR / MTORC2 regulates the barrier of chronology and spermatogenesis in mice].](https://separase.com/wp-content/uploads/2020/12/IMG-20201217-WA0088-800x600.jpg)
![[RICTOR / MTORC2 regulates the barrier of chronology and spermatogenesis in mice].](https://separase.com/wp-content/uploads/2020/12/IMG-20201217-WA0088-300x142.jpg)
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