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Temperature is one of the key environmental factors influencing crop fertility and yield. Understanding how plants sense and respond to temperature changes is, therefore, crucial for improving agricultural production. In this study, we characterized a temperature-sensitive male-sterile mutant in rice (Oryza sativa), glutamyl-tRNA synthetase 1-2 (ers1-2), that shows reduced fertility at high temperatures and restored fertility at low temperatures. Mutation of ERS1 resulted in severely delayed pollen development and meiotic progression at high temperatures, eventually leading to male sterility. Moreover, meiosis-specific events, including synapsis and crossover formation, were also delayed in ers1-2 compared with the wild type. However, these defects were all mitigated by growing ers1-2 at low temperatures. Transcriptome analysis and measurement of ascorbate, glutathione, and hydrogen peroxide (H2O2) contents revealed that the delayed meiotic progression and male sterility in ers1-2 were strongly associated with changes in reactive oxygen species (ROS) homeostasis. At high temperatures, ers1-2 exhibited decreased accumulation of ROS scavengers and overaccumulation of ROS. In contrast, at low temperatures, the antioxidant system of ROS was more active, and ROS contents were lower. These data suggest that ROS homeostasis in ers1-2 is disrupted at high temperatures but restored at low temperatures. We speculate that ERS1 dysfunction leads to changes in ROS homeostasis under different conditions, resulting in delayed or rescued meiotic progression and thermosensitive male fertility. ers1-2 may hold great potential as a thermosensitive material for crop heterosis breeding.
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During meiotic prophase I, chromosomes undergo large-scale dynamics to allow homologous chromosome pairing, prior to which chromosome ends attach to the inner nuclear envelope and form a chromosomal bouquet. Chromosome pairing is crucial for homologous recombination and accurate chromosome segregation during meiosis. However, the specific mechanism by which homologous chromosomes recognize each other is poorly understood. Here, we investigated the process of homologous chromosome pairing during early prophase I of meiosis in rice (Oryza sativa) using pooled oligo probes specific to an entire chromosome or chromosome arm. We revealed that chromosome pairing begins from both ends and extends toward the center from early zygotene through late zygotene. Genetic analysis of both trisomy and autotetraploidy also showed that pairing initiation is induced by both ends of a chromosome. However, healed ends that lack the original terminal regions on telocentric and acrocentric chromosomes cannot initiate homologous chromosome pairing, even though they may still enter the telomere clustering region at the bouquet stage. Furthermore, a chromosome that lacks the distal parts on both sides loses the ability to pair with other intact chromosomes. Thus, the native ends of chromosomes play a crucial role in initiating homologous chromosome pairing during meiosis and likely have a substantial impact on genome differentiation.
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Pareamento Cromossômico , Cromossomos de Plantas , Meiose , Oryza , Oryza/genética , Pareamento Cromossômico/genética , Cromossomos de Plantas/genética , Meiose/genética , Telômero/genética , Hibridização in Situ Fluorescente , Prófase Meiótica I/genéticaRESUMO
Crossovers (COs) are necessary for generating genetic diversity that breeders can select, but there are conserved mechanisms that regulate their frequency and distribution. Increasing CO frequency may raise the efficiency of selection by increasing the chance of integrating more desirable traits. In this study, we characterize rice FANCM and explore its functions in meiotic CO control. FANCM mutations do not affect fertility in rice, but they cause a great boost in the overall frequency of COs in both inbred and hybrid rice, according to genetic analysis of the complete set of fancm zmm (hei10, ptd, shoc1, mer3, zip4, msh4, msh5, and heip1) mutants. Although the early homologous recombination events proceed normally in fancm, the meiotic extra COs are not marked with HEI10 and require MUS81 resolvase for resolution. FANCM depends on PAIR1, COM1, DMC1, and HUS1 to perform its functions. Simultaneous disruption of FANCM and MEICA1 synergistically increases CO frequency, but it is accompanied by nonhomologous chromosome associations and fragmentations. FANCM interacts with the MHF complex, and ablation of rice MHF1 or MHF2 could restore the formation of 12 bivalents in the absence of the ZMM gene ZIP4. Our data indicate that unleashing meiotic COs by mutating any member of the FANCM-MHF complex could be an effective procedure to accelerate the efficiency of rice breeding.
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Oryza , Oryza/genética , DNA Helicases/genética , Melhoramento Vegetal , Meiose/genética , Recombinação Homóloga , Troca GenéticaRESUMO
Centromeres consist of highly repetitive sequences that are challenging to map, clone, and sequence. Active genes exist in centromeric regions, but their biological functions are difficult to explore owing to extreme suppression of recombination in these regions. In this study, we used the CRISPR/Cas9 system to knock out the transcribed gene Mitochondrial Ribosomal Protein L15 (OsMRPL15), located in the centromeric region of rice (Oryza sativa) chromosome 8, resulting in gametophyte sterility. Osmrpl15 pollen was completely sterile, with abnormalities appearing at the tricellular stage including the absence of starch granules and disrupted mitochondrial structure. Loss of OsMRPL15 caused abnormal accumulation of mitoribosomal proteins and large subunit rRNA in pollen mitochondria. Moreover, the biosynthesis of several proteins in mitochondria was defective, and expression of mitochondrial genes was upregulated at the mRNA level. Osmrpl15 pollen contained smaller amounts of intermediates related to starch metabolism than wild-type pollen, while biosynthesis of several amino acids was upregulated, possibly to compensate for defective mitochondrial protein biosynthesis and initiate consumption of carbohydrates necessary for starch biosynthesis. These results provide further insight into how defects in mitoribosome development cause gametophyte male sterility.
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Oryza , Oryza/genética , Oryza/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Genes de Plantas , Amido/metabolismo , Pólen/genética , Pólen/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Effective delivery of the CRISPR-Cas9 components is crucial to realizing the therapeutic potential. Although many delivery approaches have been developed for this application, oral delivery has not been explored due to the degradative nature of the gastrointestinal tract. For this issue, we developed a series of novel phenylboronic acid (PBA)-functionalized chitosan-polyethylenimine (CS-PEI) polymers for oral CRISPR delivery. PBA functionalization equipped the polyplex with higher stability, smooth transport across the mucus, and efficient endosomal escape and cytosolic unpackaging in the cells. From a library of 12 PBA-functionalized CS-PEI polyplexes, we identified a formulation that showed the most effective penetration in the intestinal mucosa after oral gavage to mice. The optimized formulation performed feasible CRISPR-mediated downregulation of the target protein and reduction in the downstream cholesterol. As the first oral CRISPR carrier, this study suggests the potential of addressing the needs of both local and systemic editing in a patient-compliant manner.
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Ácidos Borônicos , Quitosana , Animais , Camundongos , Polímeros , Técnicas de Transferência de GenesRESUMO
The endonuclease methyl methanesulfonate and UV-sensitive protein 81 (MUS81) has been reported to participate in DNA repair during mitosis and meiosis. However, the exact meiotic function of MUS81 in rice remains unclear. Here, we use a combination of physiological, cytological, and genetic approaches to provide evidence that MUS81 functions in atypical recombination intermediate resolution rather than crossover designation in rice. Cytological and genetic analysis revealed that the total chiasma numbers in mus81 mutants were indistinguishable from wild-type. The numbers of HEI10 foci (the sites of interference-sensitive crossovers) in mus81 were also similar to that of wild-type. Moreover, disruption of MUS81 in msh5 or msh4 msh5 background did not further decrease chiasmata frequency, suggesting that rice MUS81 did not function in crossover designation. Mutation of FANCM and ZEP1 could enhance recombination frequency. Unexpectedly, chromosome fragments and bridges were frequently observed in mus81 zep1 and mus81 fancm, illustrating that MUS81 may resolve atypical recombination intermediates. Taken together, our data suggest that MUS81 contributes little to crossover designation but plays a crucial role in the resolution of atypical meiotic intermediates by working together with other anti-crossover factors.
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Troca Genética , Oryza , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Oryza/genética , Oryza/metabolismo , Meiose/genética , Endonucleases/genética , Endonucleases/metabolismoRESUMO
RNA-dependent RNA polymerase 6 (RDR6) is a core component of the small RNA biogenesis pathway, but its function in meiosis is unclear. Here, we report a new allele of OsRDR6 (Osrdr6-meiosis [Osrdr6-mei]), which causes meiosis-specific phenotypes in rice (Oryza sativa). In Osrdr6-mei, meiotic double-strand break (DSB) formation is partially blocked. We created a biallelic mutant with more severe phenotypes, Osrdr6-bi, by crossing Osrdr6-mei with a knockout mutant, Osrdr6-edit In Osrdr6-bi meiocytes, 24 univalents were observed, and no histone H2AX phosphorylation foci were detected. Compared with the wild type, the number of 21-nucleotide small RNAs in Osrdr6-mei was dramatically lower, while the number of 24-nucleotide small RNAs was significantly higher. Thousands of differentially methylated regions (DMRs) were discovered in Osrdr6-mei, implying that OsRDR6 plays an important role in DNA methylation. There were 457 genes downregulated in Osrdr6-mei, including three genes, CENTRAL REGION COMPONENT1, P31 comet , and O. sativa SOLO DANCERS, related to DSB formation. Interestingly, the downregulated genes were associated with a high level of 24-nucleotide small RNAs but less strongly associated with DMRs. Therefore, we speculate that the alteration in expression of small RNAs in Osrdr6 mutants leads to the defects in DSB formation during meiosis, which might not be directly dependent on RNA-directed DNA methylation.
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Quebras de DNA de Cadeia Dupla , Reparo do DNA , Meiose , Oryza/genética , Proteínas de Plantas/genética , RNA Polimerase Dependente de RNA/genética , Metilação de DNA , Reparo do DNA/fisiologia , Regulação da Expressão Gênica de Plantas , Mutação , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , RNA de Plantas/biossíntese , RNA Polimerase Dependente de RNA/metabolismoRESUMO
Over the last three decades, transcranial magnetic stimulation (TMS) has gained popularity as a tool to modulate human somatosensation. However, the effects of different stimulation types on the multiple distinct subdomains of somatosensation (e.g., tactile perception, proprioception and pain) have not been systematically compared. This is especially notable in the case of newer theta-burst stimulation protocols now in widespread use. Here, we aimed to systematically and critically review the existing TMS literature and provide a complete picture of current knowledge regarding the role of TMS in modulating human somatosensation across stimulation protocols and somatosensory domains. Following the PRISMA guidelines, fifty-four studies were included in the current review and were compared based on their methodologies and results. Overall, findings from these studies provide evidence that different types of somatosensation can be both disrupted and enhanced by targeted stimulation of specific somatosensory areas. Some mixed results, however, were reported in the literature. We discussed possible reasons for these mixed results, methodological limitations of existing investigations, and potential avenues for future research.
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Percepção do Tato , Estimulação Magnética Transcraniana , Humanos , Estimulação Magnética Transcraniana/métodos , Córtex Somatossensorial/fisiologia , Percepção do Tato/fisiologia , DorRESUMO
Reliable quality control of laser welding on power batteries is an important issue due to random interference in the production process. In this paper, a quality inspection framework based on a two-branch network and conventional image processing is proposed to predict welding quality while outputting corresponding parameter information. The two-branch network consists of a segmentation network and a classification network, which alleviates the problem of large training sample size requirements for deep learning by sharing feature representations among two related tasks. Moreover, coordinate attention is introduced into feature learning modules of the network to effectively capture the subtle features of defective welds. Finally, a post-processing method based on the Hough transform is used to extract the information of the segmented weld region. Extensive experiments demonstrate that the proposed model can achieve a significant classification performance on the dataset collected on an actual production line. This study provides a valuable reference for an intelligent quality inspection system in the power battery manufacturing industry.
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The direct and rapid determination of trace cobalt ion (Co2+) in the electrolyte of zinc smelting plants is urgently needed but is impeded by the severe interference of extremely high-concentration zinc ions in the solution. Herein, colorimetric detection of Co2+ by the polyvinylpyrrolidone functionalized silver nanoparticles (PVP-AgNPs) is realized in solutions with the Zn/Co ratio being high, up to (0.8-5) × 104, which is located within the ratio range in industrial solution. The high concentration of Zn2+ induces a strong attenuation of Co2+-related signals in ultraviolet-visible (UV-vis) extinction spectra; nevertheless, a good linear range for detecting 1-6 mg/L Co2+ in 50 g/L Zn2+ solution is still acquired. The strong anti-interference toward other metal ions and the mechanism understanding for trace Co2+ detection in such a high-concentration Zn2+ solution are also revealed by systematic analysis techniques. The results extend the AgNPs as colorimetric sensors to industrial solutions, providing a new strategy for detecting trace-metal ions in industrial plants.
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Speech processing relies on interactions between auditory and motor systems and is asymmetrically organized in the human brain. The left auditory system is specialized for processing of phonemes, whereas the right is specialized for processing of pitch changes in speech affecting prosody. In speakers of tonal languages, however, processing of pitch (i.e., tone) changes that alter word meaning is left-lateralized indicating that linguistic function and language experience shape speech processing asymmetries. Here, we investigated the asymmetry of motor contributions to auditory speech processing in male and female speakers of tonal and non-tonal languages. We temporarily disrupted the right or left speech motor cortex using transcranial magnetic stimulation (TMS) and measured the impact of these disruptions on auditory discrimination (mismatch negativity; MMN) responses to phoneme and tone changes in sequences of syllables using electroencephalography (EEG). We found that the effect of motor disruptions on processing of tone changes differed between language groups: disruption of the right speech motor cortex suppressed responses to tone changes in non-tonal language speakers, whereas disruption of the left speech motor cortex suppressed responses to tone changes in tonal language speakers. In non-tonal language speakers, the effects of disruption of left speech motor cortex on responses to tone changes were inconclusive. For phoneme changes, disruption of left but not right speech motor cortex suppressed responses in both language groups. We conclude that the contributions of the right and left speech motor cortex to auditory speech processing are determined by the functional roles of acoustic cues in the listener's native language.SIGNIFICANCE STATEMENT The principles underlying hemispheric asymmetries of auditory speech processing remain debated. The asymmetry of processing of speech sounds is affected by low-level acoustic cues, but also by their linguistic function. By combining transcranial magnetic stimulation (TMS) and electroencephalography (EEG), we investigated the asymmetry of motor contributions to auditory speech processing in tonal and non-tonal language speakers. We provide causal evidence that the functional role of the acoustic cues in the listener's native language affects the asymmetry of motor influences on auditory speech discrimination ability [indexed by mismatch negativity (MMN) responses]. Lateralized top-down motor influences can affect asymmetry of speech processing in the auditory system.
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Estimulação Acústica/métodos , Percepção Auditiva/fisiologia , Idioma , Córtex Motor/fisiologia , Percepção da Fala/fisiologia , Estimulação Magnética Transcraniana/métodos , Adolescente , Adulto , Eletroencefalografia/métodos , Feminino , Humanos , Masculino , Adulto JovemRESUMO
In flowering plants, the transition from mitosis to meiosis is the precondition for gametogenesis, which is the most crucial event during sexual reproduction. Here, we report an intriguing mechanism whereby germ cells and surrounding somatic cells cooperatively involve in the meiotic switch during anther development in rice (Oryza sativa). In double mutants with loss function of both leptotene chromosome establishment- and somatic cell layer differentiation-associated genes, chromosome morphology in the reproductive cells remains the same as that in somatic cells, and sporogenous cells fail to differentiate into pollen mother cells. OsSPOROCYTELESS and MICROSPORELESS1, two pivotal genes involved in meiosis entry, are prominently downregulated in anthers of plants with mutations in both MULTIPLE SPOROCYTE1 and LEPTOTENE 1. In addition, the transcription of redox-related genes is also affected. Therefore, germ cells and the surrounding somatic cells collaboratively participate in meiosis initiation in rice.
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Cromossomos de Plantas/genética , Flores/citologia , Oryza/citologia , Proteínas de Plantas/genética , Diferenciação Celular , Flores/genética , Regulação da Expressão Gênica de Plantas , Meiose , Mutação , Oryza/genética , Células Vegetais , Plantas Geneticamente ModificadasRESUMO
Although movement variability is often attributed to unwanted noise in the motor system, recent work has demonstrated that variability may be actively controlled. To date, research on regulation of motor variability has relied on relatively simple, laboratory-specific reaching tasks. It is not clear how these results translate to complex, well-practiced tasks. Here, we test how variability is regulated during speech production, a complex, highly overpracticed, and natural motor behavior that relies on auditory and somatosensory feedback. Specifically, in a series of four experiments, we assessed the effects of auditory feedback manipulations that modulate perceived speech variability, shifting every production either toward (inward pushing) or away from (outward pushing) the center of the distribution for each vowel. Participants exposed to the inward-pushing perturbation (experiment 1) increased produced variability while the perturbation was applied as well as after it was removed. Unexpectedly, the outward-pushing perturbation (experiment 2) also increased produced variability during exposure, but variability returned to near-baseline levels when the perturbation was removed. Outward-pushing perturbations failed to reduce participants' produced variability both with larger perturbation magnitude (experiment 3) and after their variability had increased above baseline levels as a result of the inward-pushing perturbation (experiment 4). Simulations of the applied perturbations using a state-space model of motor behavior suggest that the increases in produced variability in response to the two types of perturbations may arise through distinct mechanisms. Together, these results suggest that motor variability is actively monitored and can be modulated even in complex and well-practiced behaviors such as speech.NEW & NOTEWORTHY By implementing a novel auditory feedback perturbation that modulates participants' perceived trial-to-trial variability without affecting their overall mean behavior, we show that variability in the speech motor system can be modulated. By assaying speech production, we expand our current understanding of variability to a well-practiced, complex behavior outside of the limb control system. Our results additionally highlight the need to incorporate the active control of variability in models of speech motor control.
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Percepção da Fala , Fala , Humanos , Fala/fisiologia , Percepção da Fala/fisiologia , Estimulação Acústica/métodos , Retroalimentação Sensorial/fisiologia , MovimentoRESUMO
Replication protein A (RPA), a single-stranded DNA-binding protein, plays essential role in homologous recombination. However, because deletion of RPA causes embryonic lethality in mammals, the exact function of RPA in meiosis remains unclear. In this study, we generated an rpa1a mutant using CRISPR/Cas9 technology and explored its function in rice (Oryza sativa) meiosis. In rpa1a, 12 bivalents were formed at metaphase I, just like in wild-type, but chromosome fragmentations were consistently observed at anaphase I. Fluorescence in situ hybridization assays indicated that these fragmentations were due to the failure of the recombination intermediates to resolve. Importantly, the mutant had a highly elevated chiasma number, and loss of RPA1a could completely restore the 12 bivalent formations in the zmm (for ZIP1-4, MSH4/5, and MER3) mutant background. Protein-protein interaction assays showed that RPA1a formed a complex with the methyl methansulfonate and UV sensitive 81 (and the Fanconi anemia complementation group M-Bloom syndrome protein homologs (RECQ4A)-Topoisomerase3α-RecQ-mediated genome instability 1 complex to regulate chiasma formation and processing of the recombination intermediates. Thus, our data establish a pivotal role for RPA1a in promoting the accurate resolution of recombination intermediates and in limiting redundant chiasma formation during rice meiosis.
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Proteínas de Ligação a DNA/genética , Meiose , Oryza/genética , Proteínas de Plantas/genética , Proteína de Replicação A/genética , Proteínas de Ligação a DNA/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteína de Replicação A/metabolismoRESUMO
A scheme to improve the transmittance of the metal-insulator-metal (MIM) plasmonic slot filter is proposed and numerically studied. Using this scheme, the transmittance of all channels in the MIM slot filter can be significantly improved by using only one reflector. The simulation results show that the transmittance of all channels with this scheme is almost 160% higher than without it. A single-channel filter at 980 nm and a three-channel filter are both demonstrated using this scheme. All the work above is completed by the finite-element analysis method. The model and characteristics of the structure with periodical stubs are also introduced and analyzed. For the first time, to our knowledge, the expressions of all four elements of the transfer matrix together with the reflectance of the periodical stubs are given. It is believed that our research will help to provide new ideas for improving transmittance and promote the application of plasmonic filters.
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The organization of microtubules into a bipolar spindle is essential for chromosome segregation. Both centrosome and chromatin-dependent spindle assembly mechanisms are well studied in mouse, Drosophila melanogaster, and Xenopus oocytes; however, the mechanism of bipolar spindle assembly in plant meiosis remains elusive. According to our observations of microtubule assembly in Oryza sativa, Zea mays, Arabidopsis thaliana, and Solanum lycopersicum, we propose that a key step of plant bipolar spindle assembly is the correction of the multipolar spindle into a bipolar spindle at metaphase I. The multipolar spindles failed to transition into bipolar ones in OsmtopVIB with the defect in double-strand break (DSB) formation. However, bipolar spindles were normally assembled in several other mutants lacking DSB formation, such as Osspo11-1, pair2, and crc1, indicating that bipolar spindle assembly is independent of DSB formation. We further revealed that the mono-orientation of sister kinetochores was prevalent in OsmtopVIB, whereas biorientation of sister kinetochores was frequently observed in Osspo11-1, pair2, and crc1 In addition, mutations of the cohesion subunit OsREC8 resulted in biorientation of sister kinetochores as well as bipolar spindles even in the background of OsmtopVIB Therefore, we propose that biorientation of the kinetochore is required for bipolar spindle assembly in the absence of homologous recombination.
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Meiose , Oryza/citologia , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Fuso Acromático/metabolismo , Quebras de DNA de Cadeia Dupla , Haploidia , Cinetocoros/metabolismo , Modelos Biológicos , Mutação/genéticaRESUMO
This study aims to investigate the specific mechanism of miR-139-5p regulating hepatocellular carcinoma (HCC). Bioinformatic approaches was utilized to observe miR-139-5p level in HCC and unearth its target mRNA. Next, miR-139-5p and enabled homolog (ENAH) levels in HCC cell lines and normal liver cell line were evaluated with qRT-PCR. ENAH protein level was assessed by Western blot. The cell viability, migratory and invasive capacities of HepG2 cells was observed by cell functional assays. The binding of these two genes was proved through dual-luciferase method. Xenograft nude mouse model was prepared to identify the role of miR-139-5p in vivo. Poorly expressed miR-139-5p in HCC hindered the phenotypes of cancer cells. ENAH was at high level in HCC and it is a downstream target of miR-139-5p. Additionally, ENAH could reverse the suppressive impacts of miR-139-5p on HCC cell behaviors. Likewise, miR-139-5p was determined to perform tumor-suppressing function in vivo. MiR-139-5p hampered HCC cell processes by mediating ENAH, and miR-139-5p/ENAH is hopefully to be the possible target for HCC patients.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , MicroRNAs , Camundongos , Animais , Humanos , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Regulação Neoplásica da Expressão Gênica , Proliferação de Células/genética , Linhagem Celular Tumoral , Camundongos Nus , Movimento Celular/genética , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismoRESUMO
BACKGROUND: This study aimed to explore the relationship between serum netrin-1 expression levels and acute prognosis in patients with acute ischemic stroke (AIS) within 24 hours after revascularization. METHODS: A total of 121 revascularized patients admitted to the Jinshan Branch of the Shanghai Sixth People's Hospital, China, between July 2019 and July 2021 were selected as study subjects. The primary outcome was the modified Rankin Scale (mRS) score three months after revascularization: patients with an mRS score >2 were classified into the unfavorable prognosis group and others into the favorable prognosis group. Those with serum netrin-1 expression levels greater than the median of all patients were classified into the elevated protein group and others into the decreased protein group. Multivariate logistic regression analysis was used to analyze the independent risk factors for prognosis in patients with AIS after revascularization. RESULTS: The differences between the unfavorable prognosis group and the favorable prognosis group in gender, age, coronary heart disease, and netrin-1 levels were not statistically significant (p > 0.05). However, the National Institute of Health Stroke Scale (NIHSS) scores and number of patients with comorbid hypertension in the unfavorable prognosis group were significantly higher than in the favorable prognosis group (p < 0.05). Multivariate logistic regression analysis showed that NIHSS score before revascularization was an independent risk factor for unfavorable prognosis but that netrin-1 expression levels were not significantly associated with prognosis in patients after revascularization. CONCLUSIONS: Serum netrin-1 expression levels in the acute phase are not significantly associated with prognosis in patients with AIS after revascularization.
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Isquemia Encefálica , AVC Isquêmico , Acidente Vascular Cerebral , Humanos , AVC Isquêmico/cirurgia , Netrina-1 , Isquemia Encefálica/complicações , Acidente Vascular Cerebral/complicações , China , PrognósticoRESUMO
Highly coordinated pollen wall patterning is essential for male reproductive development. Here, we report the identification of Defective Microspore Development 1 (DMD1), which encodes a nuclear-localized protein possessing transactivation activity. DMD1 is preferentially expressed in the tapetum and microspores during post-meiotic development. Mutations in DMD1 cause a male-sterile phenotype with impaired microspore cell integrity. The mutants display abnormal callose degradation, accompanied by inhibited primexine thickening in the newly released microspores. Several genes associated with callose degradation and primexine formation are downregulated in dmd1 anthers. In addition, irregular Ubisch body morphology and discontinuous endexine occur, and the baculum is completely absent in dmd1. DMD1 interacts with Tapetum Degeneration Retardation (TDR), a basic helix-loop-helix transcription factor required for exine formation. Taken together, our results suggest that DMD1 is responsible for microspore cell integrity, primexine formation and exine pattern formation during Oryza sativa (rice) microspore development.
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Oryza/metabolismo , Proteínas de Plantas/fisiologia , Pólen/metabolismo , Apoptose , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Microscopia Eletrônica de Transmissão , Oryza/crescimento & desenvolvimento , Infertilidade das Plantas , Proteínas de Plantas/metabolismo , Pólen/crescimento & desenvolvimento , Pólen/ultraestruturaRESUMO
The bipolar spindle structure in meiosis is essential for faithful chromosome segregation. PUTATIVE RECOMBINATION INITIATION DEFECT 1 (PRD1) previously has been shown to participate in the formation of DNA double strand breaks (DSBs). However, the role of PRD1 in meiotic spindle assembly has not been elucidated. Here, we reveal by both genetic analysis and immunostaining technology that PRD1 is involved in spindle assembly in rice (Oryza sativa) meiosis. We show that DSB formation and bipolar spindle assembly are disturbed in prd1 meiocytes. PRD1 signals display a dynamic pattern of localization from covering entire chromosomes at leptotene to congregating at the centromere region after leptotene. Centromeric localization of PRD1 signals depends on the organization of leptotene chromosomes, but not on DSB formation and axis establishment. PRD1 exhibits interaction and co-localization with several kinetochore components. We also find that bi-orientation of sister kinetochores within a univalent induced by mutation of REC8 can restore bipolarity in prd1. Furthermore, PRD1 directly interacts with REC8 and SGO1, suggesting that PRD1 may play a role in regulating the orientation of sister kinetochores. Taken together, we speculate that PRD1 promotes bipolar spindle assembly, presumably by modulating the orientation of sister kinetochores in rice meiosis.