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1.
Hum Reprod Open ; 2024(1): hoad051, 2024.
Article in English | MEDLINE | ID: mdl-38188854

ABSTRACT

STUDY QUESTION: What are the odds of achieving pregnancy when adopting progestin-primed ovarian stimulation (PPOS)-related protocols combined with repetitive frozen-thawed transfer (FET) cycles in patients with different clinical characteristics? SUMMARY ANSWER: The cumulative live birth rates (CLBRs) of women undergoing different PPOS-related protocols can be significantly and consistently enhanced within six FET cycles when the female age is <40 years (or even <45 years) and when >5 oocytes are retrieved, regardless of antral follicle count (AFC). WHAT IS KNOWN ALREADY: There have been numerous studies on the live birth rate of the first FET cycle in patients with PPOS-related protocols. These studies have focused mainly on comparing pregnancy outcomes with those of other stimulation protocols. However, owing to the unique features of the PPOS-related strategy, such as its flexible timing of oocyte retrieval and repeated transfer of frozen embryos, studies using the CLBR as an overall indicator of success and investigating which types of patients would benefit from this protocol are lacking. STUDY DESIGN SIZE DURATION: This retrospective cohort study included 18 593 women who underwent PPOS-related protocols (dydrogesterone + hMG, medroxyprogesterone acetate + hMG, micronized progesterone + hMG treatment, and luteal-phase ovarian stimulation protocol) from 1 March 2011 to 31 September 2022 in our centre. PARTICIPANTS/MATERIALS SETTING METHODS: The population was categorized by female age, number of oocytes retrieved, and AFC in the analysis of CLBR within six FET cycles. The age groups (Groups 1-5, respectively) were <30, 30-34, 35-39, 40-44, and ≥45 years. The number of oocytes retrieved was grouped as 1-5, 6-10, 11-15, 16-20, and >20. AFC was grouped as <5, 5-10, 11-15, and >15. The Kaplan-Meier analysis (optimistic method), which hypothesized that patients who did not continue treatment had the same chance of achieving a live birth as those who continued, and the competing risk method (conservative method) which hypothesized they had no chance of achieving a live birth, were applied. In further analyses, the Cox model and Fine-Gray model were adopted: the former corresponds to the optimistic scenario, and the latter corresponds to the pessimistic scenario. MAIN RESULTS AND THE ROLE OF CHANCE: CLBR had a declining trend with female age over six FET cycles (Groups 1-5, respectively: optimistic: 96.9%, 96.6%, 91.4%, 67.3%, and 11.7%; conservative: 87.3%, 85.0%, 74.0%, 41.3%, and 7.5%), requiring more FET cycles to achieve a success rate of at least 50% (Groups 1-5, respectively: optimistic: 2, 2, 2, 4, and >6 cycles; conservative: 2, 2, 2, >,6 and >6 cycles). CLBR showed an increasing trend with the number of oocytes retrieved (Groups 1-5, respectively: optimistic: 93.8%, 94.3%, 95.8%, 96.0%, and 95.6%; conservative: 66.2%, 78.3%, 85.6%, 88.9%, and 91.0%). All groups needed the same number of FET cycles to achieve a success rate of at least 50% (Groups 1-5, respectively: optimistic: 2, 2, 2, 2, and 2 cycles; conservative: 2, 2, 2, 2, and 2 cycles). Furthermore, the CLBR within six FET cycles had an increasing trend with AFC number (Groups 1-4, respectively: optimistic: 89.2%, 94.8%, 95.9%, and 96.3%; conservative: 67.4%, 78.2%, 83.9%, and 88.1%), with all four groups achieving a success rate of at least 50% by the second FET cycle. LIMITATIONS REASONS FOR CAUTION: The current research is limited by its retrospective design and single-centre nature, which may restrict the generalizability of our findings. WIDER IMPLICATIONS OF THE FINDINGS: This work describes two models (the Kaplan-Meier analysis and the competing risk method) to evaluate the clinical outcome of patients using PPOS-related protocols, which are especially useful for patients of advanced age or those with diminished ovarian reserve. Our findings encourage patients below 45 years old, especially younger than 40 years, and patients with lower AFCs and fewer retrieved oocytes to try this new protocol. Moreover, this study demonstrates the degree of improvement in the CLBR within six FET cycles for patients with different clinical characteristics, providing a valuable point of reference to determine whether to continue ART after a transfer failure. STUDY FUNDING/COMPETING INTERESTS: The study was supported by grants from the National Natural Science Foundation of China (82071603 to L.W., 82001502 to Y.L.). There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

2.
iScience ; 26(8): 107357, 2023 Aug 18.
Article in English | MEDLINE | ID: mdl-37520702

ABSTRACT

Progestin-primed ovarian stimulation (PPOS) is a new ovulation stimulation protocol, and its role in ovulation and regulatory mechanism is unclear. The clinical PPOS protocol was simulated in mice. The ovulated oocytes, estradiol, progesterone, and luteinizing hormone (LH) levels were analyzed at different hours after trigger. mRNA extraction and real-time PCR, hematoxylin and eosin staining, and immunofluorescence of ovaries were used to explore the involved signaling pathways. The PPOS group had a delayed ovulation at 12.5 h after trigger. Its suppressed LH level reduced the expression of luteinizing hormone/choriogonadotropin receptor (LHCGR) on the preovulatory follicles before trigger and significantly decreased the following progesterone synthesis, blood progesterone level, and progesterone receptor (PGR) expression within 4-6 h after trigger. Furthermore, the important ovulatory genes regulated by PGR including ADAMTS-1, VEGF-A, and EDN2 were downregulated, ultimately delaying the ovulation. PPOS suppresses the LH level before trigger and decreases the synthesis of progesterone after trigger, thus delaying the ovulation by downregulating the LHCGR-PGR pathway.

3.
FASEB J ; 37(5): e22927, 2023 05.
Article in English | MEDLINE | ID: mdl-37086087

ABSTRACT

miR-184 is one of the most abundant miRNAs expressed in the lens and corneal tissue. Mutations in the seed region of miR-184 are responsible for inherited anterior segment dysgenesis. Animal models recapitulating miR-184-related anterior segment dysgenesis are still lacking, and the molecular basis of ocular abnormalities caused by miR-184 dysfunction has not been well elucidated in vivo. In the present study, we constructed a miR-184-/- zebrafish line by destroying both two dre-mir-184 paralogs with CRISPR-Cas9 technology. Although there were no gross developmental defects, the miR-184-/- zebrafish displayed microphthalmia and cataract phenotypes. Cytoskeletal abnormalities, aggregation of γ-crystallin, and lens fibrosis were induced in miR-184-/- lenses. However, no obvious corneal abnormalities were observed in miR-184-/- zebrafish. Instead of apoptosis, deficiency of miR-184 led to aberrant cell proliferation and a robust increase in p21 levels in zebrafish eyes. Inhibition of p21 by UC2288 compromised the elevation of lens fibrosis markers in miR-184-/- lenses. RNA-seq demonstrated that levels of four transcriptional factors HSF4, Sox9a, CTCF, and Smad6a, all of which could suppress p21 expression, were reduced in miR-184-/- eyes. The predicted zebrafish miR-184 direct target genes (e.g., atp1a3a and nck2a) were identified and verified in miR-184-/- eye tissues. The miR-184-/- zebrafish is the first animal model mimicking miR-184-related anterior segment dysgenesis and could broaden our understanding of the roles of miR-184 in eye development.


Subject(s)
Cataract , Lens, Crystalline , MicroRNAs , Animals , Cataract/genetics , Cataract/metabolism , Lens, Crystalline/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Transcription Factors/metabolism , Zebrafish/genetics
4.
IEEE Trans Neural Netw Learn Syst ; 34(10): 7956-7966, 2023 Oct.
Article in English | MEDLINE | ID: mdl-35188893

ABSTRACT

Multimodal cross-domain sentiment analysis aims at transferring domain-invariant sentiment information across datasets to address the insufficiency of labeled data. Existing adaptation methods achieve well performance by remitting the discrepancies in characteristics of multiple modalities. However, the expressive styles of different datasets also contain domain-specific information, which hinders the adaptation performance. In this article, we propose a disentangled sentiment representation adversarial network (DiSRAN) to reduce the domain shift of expressive styles for multimodal cross-domain sentiment analysis. Specifically, we first align the multiple modalities and obtain the joint representation through a cross-modality attention layer. Then, we disentangle sentiment information from the multimodal joint representation that contains domain-specific expressive style by adversarial training. The obtained sentiment representation is domain-invariant, which can better facilitate the sentiment information transfer between different domains. Experimental results on two multimodal cross-domain sentiment analysis tasks demonstrate that the proposed method performs favorably against state-of-the-art approaches.

5.
J Clin Endocrinol Metab ; 107(4): e1467-e1476, 2022 03 24.
Article in English | MEDLINE | ID: mdl-34850010

ABSTRACT

CONTEXT: The impact of parental overweight/obese on cumulative live birth rate in in vitro fertilization/intracytoplasmic sperm injection using a freeze-all strategy is still unknown. OBJECTIVE: To explore the effect of parental body mass index (BMI) on cumulative live birth rate (CLBR) in a freeze-all strategy over 1.5 years. METHODS: This was a retrospective study in a tertiary care academic medical center; 23 482 patients (35 289 frozen-thawed embryo transfer cycles) were divided into 4 groups according to Asian BMI classification. The main outcome measure was CLBR. RESULTS: Female overweight/obesity had a lower tendency in CLBR (groups 1-4: optimistic: 69.4%, 67.9%, 62.3%, and 65.7%; conservative: 62.9%, 61.1%, 55.4%, and 57.6%) and prolonged time (groups 1-4: 11.0, 12.2, 15.9, and 13.8 months for 60% CLBR in the optimistic method; 8.7, 9.5, 11.7, 11.0 months for 50% CLBR in the conservative method). The same trend to a lesser extent was also observed in male BMI groups. When combining parental BMI, "parental overweight/obesity" had lower CLBR and longer time for reaching CLBR >50% (optimistic: 4.5 months for 60% CLBR; conservative: 3 months for 50% CLBR), followed by "only female high BMI" (optimistic: 2.1 months for 60% CLBR; conservative: 1.7 months for 50% CLBR), while "only male high BMI" had no influence. CONCLUSION: Our results showed the importance of parental BMI, female BMI, and male BMI on the 1.5-year CLBR using a freeze-all strategy; the time to reach the CLBR (60% in optimistic, 50% in conservative) for overweight and obese patients was several months, but it is not as long as losing weight.


Subject(s)
Birth Rate , Overweight , Body Mass Index , Female , Fertilization in Vitro/methods , Humans , Live Birth/epidemiology , Male , Obesity/epidemiology , Overweight/epidemiology , Ovulation Induction/methods , Pregnancy , Pregnancy Rate , Retrospective Studies
6.
IEEE Trans Image Process ; 30: 6730-6743, 2021.
Article in English | MEDLINE | ID: mdl-34283714

ABSTRACT

A simultaneous understanding of questions and images is crucial in Visual Question Answering (VQA). While the existing models have achieved satisfactory performance by associating questions with key objects in images, the answers also contain rich information that can be used to describe the visual contents in images. In this paper, we propose a re-attention framework to utilize the information in answers for the VQA task. The framework first learns the initial attention weights for the objects by calculating the similarity of each word-object pair in the feature space. Then, the visual attention map is reconstructed by re-attending the objects in images based on the answer. Through keeping the initial visual attention map and the reconstructed one to be consistent, the learned visual attention map can be corrected by the answer information. Besides, we introduce a gate mechanism to automatically control the contribution of re-attention to model training based on the entropy of the learned initial visual attention maps. We conduct experiments on three benchmark datasets, and the results demonstrate the proposed model performs favorably against state-of-the-art methods.

7.
Front Plant Sci ; 12: 800625, 2021.
Article in English | MEDLINE | ID: mdl-35126424

ABSTRACT

Glycerol-induced resistance to various pathogens has been reported in different plants. Glycerol kinase (GK), a vital rate-limiting enzyme that catalyzes glycerol conversion to glycerol-3-phosphate (G3P), participates in responses to both abiotic and biotic stresses. However, its physiological importance in rice defenses against pathogens remains unclear. In this research, quantification analysis revealed that GK levels were significantly induced in rice leaves infected by Xanthomonas oryzae pv. oryzae (Xoo) strain PXO99. A typical GK-encoding gene OsNHO1 was cloned in rice. The transcriptional levels of OsNHO1 were significantly induced by salicylic acid, jasmonic acid, and Xoo-PXO99. Ectopic expression of OsNHO1 partially rescued the resistance to P. s. pv. phaseolicola in the Arabidopsis nho1 mutant. In the overexpressing transgenic rice lines (OsNHO1-OE), the content of GK and the transcriptional level of OsNHO1 were increased and the resistance to bacterial blight and blast was improved, while reduced OsNHO1 expression impaired the resistance in OsNHO1-RNAi lines. The wax contents and expression of the wax synthesis regulatory genes were significantly increased in the overexpression lines but decreased in the OsNHO1-RNAi lines. We then confirmed the interaction partner of OsNHO1 using yeast two-hybrid and bimolecular fluorescence complementation assays. The transcription of the interaction partner-encoding genes OsSRC2 and OsPRs in OsNHO1-RNAi lines was downregulated but upregulated in OsNHO1-OE lines. Thus, we concluded that OsNHO1 provided disease resistance by affecting the wax content and modulating the transcription levels of PR genes.

8.
Front Physiol ; 11: 1102, 2020.
Article in English | MEDLINE | ID: mdl-33013471

ABSTRACT

BACKGROUND: It is unclear whether we should focus attention on cleavage-stage embryo quality and embryo development speed when transferring single particular grade vitrified-warmed blastocysts, especially poor-quality blastocysts (grade "C"). METHOD: This retrospective study considered 3386 single vitrified-warmed blastocyst transfer cycles from January 2010 to December 2017. They were divided into group 1 (AA/AB/BA, n = 374), group 2 (BB, n = 1789), group 3 (BC, n = 901), and group 4 (CB, n = 322). The effects of cleavage-stage embryo quality and embryo development speed were measured in terms of clinical pregnancy and live birth rates in each group. RESULTS: Pregnancy outcomes showed a worsening trend from groups 1 to 4; the proportion of embryos with better cleavage-stage quality and faster development speed decreased. In group 1, only the blastocyst expansion degree 3 was a negative factor in the clinical pregnancy rate (odds ratio (OR) [95% confidence interval (CI)]: 0.233 [0.091-0.595]) and live birth rate (0.280 [0.093-0.884]). In the other groups (BB, BC, and CB), blastocysts frozen on day 5 had significantly better clinical pregnancy outcomes than those frozen on day 6: 1.373 [1.095-1.722] for group 2, 1.523 [1.055-2.197] for group 3, and 3.627 [1.715-7.671] for group 4. The live birth rate was 1.342 [1.060-1.700] for group 2, 1.544 [1.058-2.253] in group 3, and 3.202 [1.509-6.795] in group 4, all Ps < 0.05). The degree of blastocoel expansion three for clinical pregnancy rate in group 2 (0.350 [0.135-0.906], P < 0.05) and day 3 blastomere number (>7) for live birth rate in group 4 (2.455 [1.190-5.063], P < 0.05) were two important factors. CONCLUSION: We should consider choosing BB/BC/CB grade blastocysts frozen on day 5, CB grade blastocysts with day 3 blastomere numbers (>7), and AA/AB/BA grade blastocysts with degrees of expansion (≥4) to obtain better pregnancy outcomes.

9.
Elife ; 92020 07 09.
Article in English | MEDLINE | ID: mdl-32644040

ABSTRACT

The gonadotropin-releasing hormone (GnRH) neurons exhibit pulse and surge modes of activity to control fertility. They also exhibit an unusual bipolar morphology comprised of a classical soma-proximal dendritic zone and an elongated secretory process that can operate as both a dendrite and an axon, termed a 'dendron'. We show using expansion microscopy that the highest density of synaptic inputs to a GnRH neuron exists at its distal dendron. In vivo, selective chemogenetic inhibition of the GnRH neuron distal dendron abolishes the luteinizing hormone (LH) surge and markedly dampens LH pulses. In contrast, inhibitory chemogenetic and optogenetic strategies targeting the GnRH neuron soma-proximal dendritic zone abolish the LH surge but have no effect upon LH pulsatility. These observations indicate that electrical activity at the soma-proximal dendrites of the GnRH neuron is only essential for the LH surge while the distal dendron represents an autonomous zone where synaptic integration drives pulsatile GnRH secretion.


Subject(s)
Dendrites/physiology , Gonadotropin-Releasing Hormone/biosynthesis , Luteinizing Hormone/antagonists & inhibitors , Animals , Dendrites/drug effects , Female , Gonadotropin-Releasing Hormone/antagonists & inhibitors , Mice , Neurons/drug effects , Neurons/physiology
10.
Arch Gynecol Obstet ; 302(2): 519-527, 2020 08.
Article in English | MEDLINE | ID: mdl-32495015

ABSTRACT

PURPOSE: To explore the trends of oocyte and pregnancy outcomes over the ovulation trigger-OPU (oocyte pickup) time interval in four mainly used COH protocols. METHODS: This retrospective study was conducted between January 2013 and July 2018. The IVF/ICSI cycles of the patients with normal ovarian reserve were included. The number of total patients was 4673, which consisted of long agonist protocol (n = 819), short agonist protocol (n = 1703), mild stimulation protocol (n = 1627), and GnRH antagonist protocol (n = 524). The primary outcome was mature oocyte rate. RESULTS: The ovulation trigger-OPU time interval and COH protocol were related to cycles with > 80% MII oocytes. Four protocols showed apparently different trends of retrieved oocyte rate and mature oocyte rate over the ovulation trigger-OPU time interval, and the long agonist protocol had the most delayed time interval than other three COH protocols in retrieving more than 60% oocytes (35.4-39.6 h vs. 34.6-38.6 h vs. 32.5-37.5 h vs. 33.8-37.7 h) and getting more than 80% mature oocytes (35.0-39.7 h vs. 36.0-37.7 h vs. 34.1-35.5 h vs. 34.5-36.3 h). And the adjusted odds ratio (OR) of the cumulative live birth rate (CLBR) (OR 1.360, 95% Confidence Interval (CI) 1.156-1.549, P < 0.05) significantly increased with the trigger-OPU time interval in the long agonist protocol. CONCLUSIONS: For getting more and mature oocytes, the ovulation trigger-OPU time intervals should be gradually prolonged from the mild stimulation protocol, the GnRH antagonist protocol, and the short protocol to the long agonist protocol. And the prolonged ovulation trigger-OPU time interval in the long agonist protocol brings higher live birth rate (LBR) and CLBR.


Subject(s)
Ovulation Induction/methods , Female , Humans , Ovary , Pregnancy , Pregnancy Outcome , Retrospective Studies
11.
Biochim Biophys Acta Mol Basis Dis ; 1866(5): 165724, 2020 05 01.
Article in English | MEDLINE | ID: mdl-32061775

ABSTRACT

Removal of nuclei in lens fiber cells is required for organelle-free zone (OFZ) formation during lens development. Defect in degradation of nuclear DNA leads to cataract formation. DNase2ß degrades nuclear DNA of lens fiber cells during lens differentiation in mouse. Hsf4 is the principal heat shock transcription factor in lens and facilitates the lens differentiation. Knockout of Hsf4 in mouse and zebrafish resulted in lens developmental defect that was characterized by retaining of nuclei in lens fiber cells. In previous in vitro studies, we found that Hsf4 promoted DNase2ß expression in human and mouse lens epithelial cells. In this study, it was found that, instead of DNase2ß, DNase1l1l is uniquely expressed in zebrafish lens and was absent in Hsf4-/- zebrafish lens. Using CRISPR-Cas9 technology, a DNase1l1l knockout zebrafish line was constructed, which developed cataract. Deletion of DNase1l1l totally abrogated lens primary and secondary fiber cell denucleation process, whereas had little effect on the clearance of other organelles. The transcriptional regulation of DNase1l1l was dramatically impaired in Hsf4-/- zebrafish lens. Rescue of DNase1l1l mRNA into Hsf4-/- zebrafish embryos alleviated its defect in lens fiber cell denucleation. Our results in vivo demonstrated that DNase1l1l is the primary DNase responsible for nuclear DNA degradation in lens fiber cells, and Hsf4 can transcriptionally activate DNase1l1l expression in zebrafish.


Subject(s)
Cataract/genetics , Deoxyribonucleases/genetics , Gene Expression Regulation, Developmental , Heat Shock Transcription Factors/metabolism , Lens, Crystalline/embryology , Zebrafish Proteins/genetics , Animals , Animals, Genetically Modified , CRISPR-Cas Systems/genetics , Cataract/pathology , Cell Nucleus/metabolism , Deoxyribonucleases/metabolism , Disease Models, Animal , Embryo, Nonmammalian , Epithelial Cells/cytology , Epithelial Cells/metabolism , Female , Gene Knockout Techniques , Heat Shock Transcription Factors/genetics , Humans , Lens, Crystalline/cytology , Lens, Crystalline/metabolism , Lens, Crystalline/pathology , Male , Zebrafish , Zebrafish Proteins/metabolism
12.
Article in English | MEDLINE | ID: mdl-31681169

ABSTRACT

Background: To investigate the optimal ovulation trigger-oocyte pickup (OPU) interval of a progestin-primed ovarian stimulation (PPOS) protocol. Method: Patients with normal ovarian reserve in their first PPOS OPU cycle were enrolled in this retrospective cohort study between July 2013 and April 2018. This retrospective cohort study included two parts. In part I, we studied the regression trend of mature oocyte rate, implantation rate, and live birth rate within the whole ovulation trigger-OPU interval of 7,258 patients. To homogenize some clinical characters that were key regulators of OPU time, in part II, we used propensity score matching to auto-select patients among trigger-OPU interval group 1 (35.6-36.4 h), group 2 (36.4-37.1 h), and group 3 (37.1-37.8 h) and analyzed clinical outcomes. Results: Study part I showed that the whole ovulation trigger-OPU interval (33-39.5 h) of PPOS protocol had a trend of a high mature oocyte rate (>80%), increasing implantation rate, and high live birth rate. Propensity score matching of patients with homogeneous clinical characteristics further indicated that the trigger-OPU interval within groups 2 and 3 (36.4-37.8 h) had significantly higher mature oocyte rates (84.54% vs. 84.60% vs. 82.34%, P = 0.002) and implantation rates (34.17% vs. 34.37% vs. 29.61%, P < 0.05) than group 1. The same tend was observed in the live birth rate. Conclusions: The ovulation trigger-OPU interval of 36.4-37.8 h is optimal for most patients using a PPOS protocol.

13.
Gene ; 707: 86-92, 2019 Jul 30.
Article in English | MEDLINE | ID: mdl-31071385

ABSTRACT

Retinitis pigmentosa (RP) is the most common form of inherited retinal degenerative diseases. X-linked RP accounts for nearly 15% of all RP cases. In this study, we identified a novel RP2 missense mutation Q158P in a Chinese XLRP family. The RP2 Q158P mutation located in the RP2 TBCC domain and obviously destabilized RP2 protein in ARPE-19 cells. The proteasome inhibitor MG132 could restore the RP2 Q158P protein levels. Meanwhile, lower doses of bortezomib and carfilzomib, another two proteasome inhibitors that have been approved in multiple myeloma clinical therapy, also could rescue the RP2 Q158P protein levels. The ubiquitination of RP2 Q158P protein obviously increased when compared with wild type RP2 protein. Our findings broadened the spectrum of RP2 mutations and may contribute a better understanding of the molecular mechanism of XLRP.


Subject(s)
Eye Proteins/chemistry , Eye Proteins/genetics , Genetic Diseases, X-Linked/genetics , Intracellular Signaling Peptides and Proteins/chemistry , Intracellular Signaling Peptides and Proteins/genetics , Membrane Proteins/chemistry , Membrane Proteins/genetics , Mutation, Missense , Retinitis Pigmentosa/genetics , Cell Line , China , DNA Mutational Analysis , Female , GTP-Binding Proteins , Humans , Male , Models, Molecular , Pedigree , Protein Domains , Protein Stability , Sequence Analysis, DNA
14.
Yi Chuan ; 39(1): 56-65, 2017 01 20.
Article in English | MEDLINE | ID: mdl-28115306

ABSTRACT

LEAFY/FLORICAULA (LFY/FLO) is a family of plant-specific transcription factors, which plays an important role(s) in the regulation of floral organ formation and development. So far, LFY regulation on floral development in wild soybean has not been reported in the literature. In this study, the LFY gene, GsLFY, has been isolated from Glycine soja, and characterized with molecular and transgenic techniques. The cDNA for GsLFY gene is 1224 bp in length and contains an open reading frame encoding a polypeptide of 407 amino acids. Quantitative RT-PCR analysis shows that GsLFY is prominently expressed in various tissues, including roots, flowers and seeds. Among the four floral organs, GsLFY is expressed highly in sepals and stamens while weakly in the petals and carpels. Yeast two-hybrid experiments show that GsLFY possesses transactivation activity while transient expression analysis with Arabidopsis thaliana protoplasts shows that GsLFY protein is localized in the nucleus, supporting the notion that GsLFY is a transcription factor. The GsLFY transgenic tobacco plants flower about 29 days earlier than the wild-type plants, thereby providing a potential rationale for developing new soybean varieties with altered flowering time.


Subject(s)
Fabaceae/genetics , Flowers/genetics , Genes, Plant/genetics , Plant Proteins/genetics , Amino Acid Sequence , Sequence Alignment
15.
Mol Plant Pathol ; 17(3): 412-26, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26123657

ABSTRACT

Several plant lipid transfer proteins (LTPs) act positively in plant disease resistance. Here, we show that LTP3 (At5g59320), a pathogen and abscisic acid (ABA)-induced gene, negatively regulates plant immunity in Arabidopsis. The overexpression of LTP3 (LTP3-OX) led to an enhanced susceptibility to virulent bacteria and compromised resistance to avirulent bacteria. On infection of LTP3-OX plants with Pseudomonas syringae pv. tomato, genes involved in ABA biosynthesis, NCED3 and AAO3, were highly induced, whereas salicylic acid (SA)-related genes, ICS1 and PR1, were down-regulated. Accordingly, in LTP3-OX plants, we observed increased ABA levels and decreased SA levels relative to the wild-type. We also showed that the LTP3 overexpression-mediated enhanced susceptibility was partially dependent on AAO3. Interestingly, loss of function of LTP3 (ltp3-1) did not affect ABA pathways, but resulted in PR1 gene induction and elevated SA levels, suggesting that LTP3 can negatively regulate SA in an ABA-independent manner. However, a double mutant consisting of ltp3-1 and silent LTP4 (ltp3/ltp4) showed reduced susceptibility to Pseudomonas and down-regulation of ABA biosynthesis genes, suggesting that LTP3 acts in a redundant manner with its closest homologue LTP4 by modulating the ABA pathway. Taken together, our data show that LTP3 is a novel negative regulator of plant immunity which acts through the manipulation of the ABA-SA balance.


Subject(s)
Abscisic Acid/biosynthesis , Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Arabidopsis/microbiology , Aldehyde Oxidase/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Disease Resistance , Disease Susceptibility , Gene Expression Regulation, Plant , Genes, Plant , Germination/genetics , Mutation/genetics , Plant Diseases/microbiology , Pseudomonas syringae/pathogenicity , Seeds/genetics , Signal Transduction , Virulence
16.
Can J Physiol Pharmacol ; 92(8): 655-60, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24988216

ABSTRACT

This study aimed to investigate the effect of baclofen, a γ-aminobutyric acid B (GABAB) receptor agonist, on the expression of p-CREB and NR2B in the spinal dorsal horn of rats with diabetic neuropathic pain (DNP). The DNP rats, which were successfully induced with streptozocin, were distributed among 3 groups that were treated with saline (D1 group), baclofen (D2 group), or CGP55845 + baclofen (D3 group) continuously for 4 days. The rats induced with saline and subsequently treated with saline were used as controls (C group). The times for the paw withdrawal threshold and thermal withdrawal latency of the D1 group were lower than those for the C group, and were significantly increased after baclofen treatment, but not when GABA receptor was pre-blocked with CGP55845 (D3 group). Increased protein expression levels of NR2B and p-CREB and mRNA levels of NR2B were found in the D1 group when compared with the controls. Baclofen treatment significantly suppressed their expression, bringing it close to the levels of controls. However, in the D3 group, the expression of p-CREB and NR2B were still significantly higher than that of the controls. Activation of GABAB receptor by baclofen attenuates diabetic neuropathic pain, which may partly be accomplished via down-regulating the expression of p-CREB and NR2B.


Subject(s)
Baclofen/pharmacology , CREB-Binding Protein/metabolism , Diabetic Neuropathies/metabolism , GABA-B Receptor Agonists/pharmacology , Neuralgia/drug therapy , Receptors, N-Methyl-D-Aspartate/metabolism , Spinal Cord Dorsal Horn/drug effects , Animals , Baclofen/therapeutic use , CREB-Binding Protein/genetics , Diabetic Neuropathies/physiopathology , GABA-B Receptor Agonists/therapeutic use , GABA-B Receptor Antagonists/pharmacology , Gene Expression , Injections, Spinal , Male , Neuralgia/metabolism , Neuralgia/physiopathology , Pain Measurement , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/genetics , Spinal Cord Dorsal Horn/metabolism
17.
J Neurol Sci ; 341(1-2): 68-72, 2014 Jun 15.
Article in English | MEDLINE | ID: mdl-24787504

ABSTRACT

N-methyl-D-aspartate receptor (NMDAR) activity is increased, while GABAB receptor is downregulated in the spinal cord dorsal horn in diabetic neuropathy. In this study, we determined the interaction of NMDARs and GABAB receptors in streptozotocin (STZ)-induced diabetic neuropathy. The paw withdrawal threshold (PWT) was significantly lower in STZ-treated rats than in vehicle-treated rats. Intrathecal injection of baclofen, a GABAB receptor agonist, significantly increased the PWT in STZ-treated rats, an effect that was abolished by pre-administration of the GABAB receptor specific antagonist CGP55845. Spinal NR2B, an NMDA receptor subunit, protein and mRNA expression levels were significantly higher in STZ-treated rats than in vehicle-treated rats. Intrathecal baclofen significantly reduced the NR2B protein and mRNA expression levels in STZ-treated rats. Intrathecal administration of CGP55845 eliminated baclofen-induced reduction of NR2B protein and mRNA levels in STZ-treated rats. In addition, the phosphorylated cAMP response element-binding (CREB) protein level was significantly higher in the spinal cord dorsal horn in STZ-treated rats compared with vehicle-treated rats. Intrathecal injection of baclofen significantly decreased phosphorylated CREB protein level in STZ-treated rats; an effect was blocked by CGP55845. These data suggest that activation of GABAB receptors in the spinal cord dorsal horn normalizes NMDAR expression level in diabetic neuropathic pain.


Subject(s)
Diabetic Neuropathies/pathology , Receptors, GABA-B/metabolism , Receptors, N-Methyl-D-Aspartate/metabolism , Spinal Cord/metabolism , Analysis of Variance , Animals , Antibiotics, Antineoplastic/toxicity , Baclofen/pharmacology , CREB-Binding Protein/genetics , CREB-Binding Protein/metabolism , Diabetic Neuropathies/chemically induced , Disease Models, Animal , GABA Antagonists/pharmacology , GABA-B Receptor Agonists/pharmacology , Gene Expression Regulation/drug effects , Male , Pain Measurement/drug effects , Phosphinic Acids/pharmacology , Propanolamines/pharmacology , Rats , Rats, Sprague-Dawley , Receptors, GABA-B/genetics , Receptors, N-Methyl-D-Aspartate/genetics , Spinal Cord/drug effects , Streptozocin/toxicity , Time Factors
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