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Zhongguo Zhen Jiu = Chinese Acupuncture... Jun 2024To assess the effect of acupuncture therapy (acupuncture for regulating menstruation and promoting pregnancy) on pregnancy outcomes in patients with diminished ovarian...
OBJECTIVE
To assess the effect of acupuncture therapy (acupuncture for regulating menstruation and promoting pregnancy) on pregnancy outcomes in patients with diminished ovarian reserve (DOR) undergoing in vitro fertilization-embryo transfer (IVF-ET).
METHODS
Eighty women with DOR were divided into an observation group (40 cases, 1 case dropped out) and a control group (40 cases, 1 case dropped out) according to whether acupuncture therapy was given or not. In the control group, IVF-ET was delivered. In the observation group, before IVF-ET, acupuncture therapy was given. Two groups of acupoints were used alternatively, including Baihui (GV 20), Shenting (GV 24), Benshen (GB 13), Zhongwan (CV 12), Guanyuan (CV 4), and bilateral Tianshu (ST 25), Shenshu (BL 23), Ciliao (BL 32), etc. Acupuncture was operated once every other day, three interventions a week, for 12 weeks. The primary outcome was clinical pregnancy rate (CPR). Secondary outcomes included the total days and amount of gonadotropin (Gn) used, the number of oocytes retrieved, the number of oocytes in metaphase of second meiosis (MⅡ), the number of transferable embryos, the number of high-quality embryos, the cycle cancellation rate, the positive rate of human choriogonadotropin (HCG), the embryo implantation rate, live birth rate (LBR), the basal serum levels of sex hormones (follicular stimulating hormone [FSH], estradiol (E2), FSH/luteinizing hormone [LH]) and antral follicle count (AFC).
RESULTS
CPR in the observation group was higher than that in the control group (53.8% [21/39] . 17.9% [7/39], <0.05). The results of the number of oocytes retrieved, the number of oocytes in MⅡ, the number of transferable embryos, the number of high-quality embryos, the positive rate of HCG, the embryo implantation rate, and LBR in the observation group were higher than those in the control group (<0.05). The serum level of FSH and FSH/LH in the observation group were lower thau those in the control group (<0.05). The differences were not significant statistically in the total days and amount of Gn used, the cycle cancellation rate, serum level of E2 and AFC between the two groups (>0.05). Logic regression analysis showed that CPR increased in the observation group when compared with that of the control group ( = 5.33, 95%: 1.90-14.97, = 0.001).
CONCLUSION
Acupuncture can improve the pregnancy outcomes of DOR women undergoing IVF-ET.
Topics: Humans; Female; Adult; Pregnancy; Fertilization in Vitro; Acupuncture Therapy; Ovarian Reserve; Embryo Transfer; Pregnancy Outcome; Infertility, Female; Cohort Studies; Acupuncture Points; Pregnancy Rate; Young Adult
PubMed: 38867627
DOI: 10.13703/j.0255-2930.20230716-k0002 -
Scientific Reports Jun 2024CRISPR-Cas9 technology has facilitated development of strategies that can potentially provide more humane and effective methods to control invasive vertebrate species,...
CRISPR-Cas9 technology has facilitated development of strategies that can potentially provide more humane and effective methods to control invasive vertebrate species, such as mice. One promising strategy is X chromosome shredding which aims to bias offspring towards males, resulting in a gradual and unsustainable decline of females. This method has been explored in insects with encouraging results. Here, we investigated this strategy in Mus musculus by targeting repeat DNA sequences on the X chromosome with the aim of inducing sufficient DNA damage to specifically eliminate X chromosome-bearing sperm during gametogenesis. We tested three different guide RNAs (gRNAs) targeting different repeats on the X chromosome, together with three male germline-specific promoters for inducing Cas9 expression at different stages of spermatogenesis. A modest bias towards mature Y-bearing sperm was detected in some transgenic males, although this did not translate into significant male-biasing of offspring. Instead, cleavage of the X chromosome during meiosis typically resulted in a spermatogenic block, manifest as small testes volume, empty tubules, low sperm concentration, and sub/infertility. Our study highlights the importance of controlling the timing of CRISPR-Cas9 activity during mammalian spermatogenesis and the sensitivity of spermatocytes to X chromosome disruption.
Topics: Animals; Male; Mice; CRISPR-Cas Systems; Spermatogenesis; X Chromosome; Female; RNA, Guide, CRISPR-Cas Systems; Spermatozoa; Mice, Transgenic; Meiosis
PubMed: 38866815
DOI: 10.1038/s41598-024-63706-4 -
Cell Stem Cell Jun 2024
PubMed: 38866014
DOI: 10.1016/j.stem.2024.06.005 -
Proceedings of the National Academy of... Jun 2024Meiosis, a reductional cell division, relies on precise initiation, maturation, and resolution of crossovers (COs) during prophase I to ensure the accurate segregation...
Meiosis, a reductional cell division, relies on precise initiation, maturation, and resolution of crossovers (COs) during prophase I to ensure the accurate segregation of homologous chromosomes during metaphase I. This process is regulated by the interplay of RING-E3 ligases such as RNF212 and HEI10 in mammals. In this study, we functionally characterized a recently identified RING-E3 ligase, RNF212B. RNF212B colocalizes and interacts with RNF212, forming foci along chromosomes from zygonema onward in a synapsis-dependent and DSB-independent manner. These consolidate into larger foci at maturing COs, colocalizing with HEI10, CNTD1, and MLH1 by late pachynema. Genetically, RNF212B foci formation depends on but not on , and , while the unloading of RNF212B at the end of pachynema is dependent on and . Mice lacking RNF212B, or expressing an inactive RNF212B protein, exhibit modest synapsis defects, a reduction in the localization of pro-CO factors (MSH4, TEX11, RPA, MZIP2) and absence of late CO-intermediates (MLH1). This loss of most COs by diakinesis results in mostly univalent chromosomes. Double mutants for and exhibit an identical phenotype to that of single mutants, while double heterozygous demonstrate a dosage-dependent reduction in CO number, indicating a functional interplay between paralogs. SUMOylome analysis of testes from mutants and pull-down analysis of Sumo- and Ubiquitin-tagged HeLa cells, suggest that RNF212B is an E3-ligase with Ubiquitin activity, serving as a crucial factor for CO maturation. Thus, RNF212 and RNF212B play vital, yet overlapping roles, in ensuring CO homeostasis through their distinct E3 ligase activities.
Topics: Animals; Mice; Meiosis; Male; Female; Ubiquitin-Protein Ligases; Crossing Over, Genetic; Chromosome Pairing; Poly-ADP-Ribose Binding Proteins; Mice, Knockout; Humans; Ligases
PubMed: 38865271
DOI: 10.1073/pnas.2320995121 -
Molecular Biology of the Cell Jun 2024The reductional division of meiosis I requires the separation of chromosome pairs towards opposite poles. We have previously implicated the outer kinetochore protein...
The reductional division of meiosis I requires the separation of chromosome pairs towards opposite poles. We have previously implicated the outer kinetochore protein SPC105R/KNL1 in driving meiosis I chromosome segregation through lateral attachments to microtubules and co-orientation of sister centromeres. To identify the domains of SPC105R that are critical for meiotic chromosome segregation, an RNAi-resistant gene expression system was developed. We found that the SPC105R C-terminal domain (aa 1284-1960) is necessary and sufficient for recruiting NDC80 to the kinetochore and building the outer kinetochore. Furthermore, the C-terminal domain recruits BUBR1, which in turn recruits the cohesion protection proteins MEI-S332 and PP2A. Of the remaining 1283 amino acids, we found the first 473 are most important for meiosis. The first 123 amino acids of the N-terminal half of SPC105R contain the conserved SLRK and RISF motifs that are targets of PP1 and Aurora B kinase and are most important for regulating the stability of microtubule attachments and maintaining metaphase I arrest. The region between amino acids 124 and 473 are required for lateral microtubule attachments and bi-orientation of homologs, which are critical for accurate chromosome segregation in meiosis I.
PubMed: 38865189
DOI: 10.1091/mbc.E24-02-0067 -
ELife Jun 2024Cohesin is a multi-subunit protein that plays a pivotal role in holding sister chromatids together during cell division. Sister chromatid cohesion 3 (SCC3), constituents...
Cohesin is a multi-subunit protein that plays a pivotal role in holding sister chromatids together during cell division. Sister chromatid cohesion 3 (SCC3), constituents of cohesin complex, is highly conserved from yeast to mammals. Since the deletion of individual cohesin subunit always causes lethality, it is difficult to dissect its biological function in both mitosis and meiosis. Here, we obtained weak mutants using CRISPR-Cas9 system to explore its function during rice mitosis and meiosis. The weak mutants displayed obvious vegetative defects and complete sterility, underscoring the essential roles of SCC3 in both mitosis and meiosis. SCC3 is localized on chromatin from interphase to prometaphase in mitosis. However, in meiosis, SCC3 acts as an axial element during early prophase I and subsequently situates onto centromeric regions following the disassembly of the synaptonemal complex. The loading of SCC3 onto meiotic chromosomes depends on REC8. shows severe defects in homologous pairing and synapsis. Consequently, SCC3 functions as an axial element that is essential for maintaining homologous chromosome pairing and synapsis during meiosis.
Topics: Chromosome Pairing; Meiosis; Cell Cycle Proteins; Oryza; Chromosomal Proteins, Non-Histone; Cohesins; Mitosis; Synaptonemal Complex; CRISPR-Cas Systems
PubMed: 38864853
DOI: 10.7554/eLife.94180 -
BMC Plant Biology Jun 2024Cytoplasmic male sterility (CMS) has greatly improved the utilization of heterosis in crops due to the absence of functional male gametophyte. The newly developed...
BACKGROUND
Cytoplasmic male sterility (CMS) has greatly improved the utilization of heterosis in crops due to the absence of functional male gametophyte. The newly developed sporophytic D1 type CMS (CMS-D1) rice exhibits unique characteristics compared to the well-known sporophytic CMS-WA line, making it a valuable resource for rice breeding.
RESULTS
In this research, a novel CMS-D1 line named Xingye A (XYA) was established, characterized by small, transparent, and shriveled anthers. Histological and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assays conducted on anthers from XYA and its maintainer line XYB revealed that male sterility in XYA is a result of delayed degradation of tapetal cells and abnormal programmed cell death (PCD) of microspores. Transcriptome analysis of young panicles revealed that differentially expressed genes (DEGs) in XYA, compared to XYB, were significantly enriched in processes related to chromatin structure and nucleosomes during the microspore mother cell (MMC) stage. Conversely, processes associated with sporopollenin biosynthesis, pollen exine formation, chitinase activity, and pollen wall assembly were enriched during the meiosis stage. Metabolome analysis identified 176 specific differentially accumulated metabolites (DAMs) during the meiosis stage, enriched in pathways such as α-linoleic acid metabolism, flavone and flavonol biosynthesis, and linolenic acid metabolism. Integration of transcriptomic and metabolomic data underscored the jasmonic acid (JA) biosynthesis pathway was significant enriched in XYA during the meiosis stage compared to XYB. Furthermore, levels of JA, MeJA, OPC4, OPDA, and JA-Ile were all higher in XYA than in XYB at the meiosis stage.
CONCLUSIONS
These findings emphasize the involvement of the JA biosynthetic pathway in pollen development in the CMS-D1 line, providing a foundation for further exploration of the molecular mechanisms involved in CMS-D1 sterility.
Topics: Oryza; Pollen; Plant Infertility; Transcriptome; Gene Expression Profiling; Metabolomics; Metabolome; Gene Expression Regulation, Plant; Meiosis
PubMed: 38862889
DOI: 10.1186/s12870-024-05259-2 -
DNA Repair Jun 2024To identify new molecular components of the Brh2-governed homologous recombination (HR)-network in the highly radiation-resistant fungus Ustilago maydis, we undertook a...
To identify new molecular components of the Brh2-governed homologous recombination (HR)-network in the highly radiation-resistant fungus Ustilago maydis, we undertook a genetic screen for suppressors of blm- hydroxyurea (HU)-sensitivity. Twenty DNA-damage sensitive mutants were obtained, three of which showing slow-growth phenotypes. Focusing on the "normally" growing candidates we identified five mutations, two in previously well-defined genes (Rec2 and Rad51) and the remaining three in completely uncharacterized genes (named Rec3, Bls9 and Zdr1). A common feature among these novel factors is their prominent role in DNA repair. Rec3 contains the P-loop NTPase domain which is most similar to that found in U. maydis Rec2 protein, and like Rec2, Rec3 plays critical roles in induced allelic recombination, is crucial for completion of meiosis, and with regard to DNA repair Δrec3 and Δrec2 are epistatic to one another. Importantly, overexpression of Brh2 in Δrec3 can effectively restore DNA-damage resistance, indicating a close functional connection between Brh2 and Rec3. The Bls9 does not seem to have any convincing domains that would give a clue as to its function. Nevertheless, we present evidence that, besides being involved in DNA-repair, Bls9 is also necessary for HR between chromosome homologs. Moreover, Δbls9 showed epistasis with Δbrh2 with respect to killing by DNA-damaging agents. Both, Rec3 and Bls9, play an important role in protecting the genome from mutations. Zdr1 is Cys2-His2 zinc finger (C2H2-ZF) protein, whose loss does not cause a detectable change in HR. Also, the functions of both Bls9 and Zdr1 genes are dispensable in meiosis and sporulation. However, Zdr1 appears to have overlapping activities with Blm and Mus81 in protecting the organism from methyl methanesulfonate- and diepoxybutane-induced DNA-damage. Finally, while deletion of Rec3 and Zdr1 can suppress HU-sensitivity of blm-, Δgen1, and Δmus81 mutants, interestingly loss of Bls9 does not rescue HU-sensitivity of Δgen1.
PubMed: 38861762
DOI: 10.1016/j.dnarep.2024.103709 -
BioRxiv : the Preprint Server For... May 2024Meiotic recombination is required for faithful chromosome segregation in most sexually reproducing organisms and shapes the distribution of genetic variation in...
Meiotic recombination is required for faithful chromosome segregation in most sexually reproducing organisms and shapes the distribution of genetic variation in populations. Both the overall rate and the spatial distribution of crossovers vary within and between species. Adjacent crossovers on the same chromosome tend to be spaced more evenly than expected at random, a phenomenon known as crossover interference. Although interference has been observed in many taxa, the factors that influence the strength of interference are not well understood. We used house mice (), a well-established model system for understanding recombination, to study the effects of genetics and age on recombination rate and interference in the male germline. We analyzed crossover positions in 503 progeny from reciprocal F1 hybrids between inbred strains representing the three major subspecies of house mice. Consistent with previous studies, autosomal alleles from tend to increase recombination rate, while inheriting a X chromosome decreases recombination rate. Old males transmit an average of 0.6 more crossovers per meiosis (5.0%) than young males, though the effect varies across genetic backgrounds. We show that the strength of crossover interference depends on genotype, providing a rare demonstration that interference evolves over short timescales. Differences between reciprocal F1s suggest that X-linked factors modulate the strength of interference. Our findings motivate additional comparisons of interference among recently diverged species and further examination of the role of paternal age in determining the number and positioning of crossovers.
PubMed: 38854148
DOI: 10.1101/2024.05.28.596233 -
BioRxiv : the Preprint Server For... May 2024An model of human meiosis would accelerate research into this important reproductive process and development of therapies for infertility. We have developed a method to...
An model of human meiosis would accelerate research into this important reproductive process and development of therapies for infertility. We have developed a method to induce meiosis starting from male or female human pluripotent stem cells. We demonstrate that DNMT1 inhibition, retinoid signaling activation, and overexpression of regulatory factors (anti-apoptotic BCL2, and pro-meiotic HOXB5, BOLL, or MEIOC) rapidly activates meiosis, with leptonema beginning at 6 days, zygonema at 9 days, and pachynema at 12 days. Immunofluorescence microscopy shows key aspects of meiosis, including chromosome synapsis and sex body formation. The meiotic cells express genes similar to meiotic oogonia , including all synaptonemal complex components and machinery for meiotic recombination. These findings establish an accessible system for inducing human meiosis .
PubMed: 38854076
DOI: 10.1101/2024.05.31.596483