SCIENTIA HORTICULTURAE
·
2023
·
IF 4.3
Yuan Zhong,Ming-jie Du,Run-ze Ji,Fang-yun Cheng
Polyploid breeding is one of the most important approaches to improve the traits of cultivated plants, but there has been no feasible method to get polyploid tree peony. In this study, we revealed a sexual triploidization approach from a diploid intersubsectional hybrid tree peony, Paeonia × lemoinei ‘High Noon’, to an allotriploid one, P. × lemoinei ‘Oukan’, by accurate karyotyping based on genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH). The ancestor genomes from P. suffruticosa and P. delavayi , which were defined as A-genome and B-genome respectively, were clearly distinguished by GISH in ‘High Noon’ and ‘Oukan’. Nonhomologous chromosomes were successfully recognized by means of 5S and 45S rDNA FISH. Karyotype comparation based on GISH and FISH indicated that the B-genome and one of the two A-genomes in ‘Oukan’ (2 n =3 x =15, AAB) were highly consistent to the corresponding genomes in ‘High Noon’ (2 n =2 x =10, AB), implying the triploid origination via 2 n gametes produced by first division restitution (FDR). In addition, it was demonstrated for the first time that the dual in series sites of 5S rDNA located on the short arm of chromosome 3 in both A and B-genomes. Besides the familiar strong 45S rDNA signals observed at terminal region of short arms, single weak 45S rDNA signal was newly detected at terminal region in the short arm of certain chromosome 1 in A-genome, and at interstitial-distal region in the long arm of chromosome 5 in both genomes. These findings provide a proof and theoretical support for tree peony polyploid breeding, and an accurate karyotyping method for genetic research of plants in Paeonia .
研究领域:
植物学
·
遗传学
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DOI
高影响因子
JOURNAL OF COMPARATIVE NEUROLOGY
·
2022
·Vol. 531
·期 2
·294-313
·
IF 3.028
Xulin Li,Gaoyang Zhao,Hongren Huang,Jialin Ye,Junfeng Xu,Yuan Zhou,Xiaojuan Zhu,Liping Wang,Feng Wang
Many brain functions that underlie behavior, cognition, and emotions vary with age, as does susceptibility to neuropsychological disorders. The expression of specific genes that are involved in these functions, such as the genes encoding for oxytocin, its receptors, and apolipoprotein D, varies with age across different brain regions. The cannabinoid 1 receptor (CB 1 R) is one of the most widely spread G-protein coupled receptors in the central nervous system and is increasingly recognized for its important contribution to various brain functions. Although changes in CB 1 R expression with age have been reported in the male mouse brain, they have not been well investigated in the female brain. Here, we used fluorescence in situ hybridization to target CB 1 R mRNA in the whole brains of female C57BL/6J mice aged 4, 6, 12, 52 (12 months) and 86 weeks (20 months), and quantified CB 1 R-positive cells in 36 brain regions across the whole brain. The results showed that CB 1 R-positive cells number changed with age. Specifically, CB 1 R expression increased with age in some subregions of the cortex, decreased with age in the lateral septal area, and reached its lowest level at 52 weeks in the thalamus, hypothalamus, and hindbrain subregions. Cluster analysis revealed that some brain regions shared similar temporal characteristics in CB 1 R-positive cell number across the lifespan. Our results provide evidence that investigation of the neural basis of age-related characteristics of female brain functions is not only warranted but required.
研究领域:
神经科学
·
分子生物学
·
衰老
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Hui Yang,Yang Gao,Siqi Wang,You Qin,Lu Xu,Dan Jin,Fan Yang,Guo-Jun Zhang
In this study, we report an enzyme-free and conjugation-free electrochemical genosensor enabling an ultrasensitive readout of BRCA-1, a breast cancer susceptibility gene. The sensor employs a target-responsive hybridization chain reaction (HCR) to significantly amplify the detectable current signals. By means of a functional auxiliary probe pair and a versatile initiator sequence, a linear DNA concatemer structure can be formed via spontaneous and continuous polymerization of DNA oligomers in the presence of target sequence. Such a DNA nanoassembly endows the genosensor an ultrahigh sensitivity up to 1 aM, which is higher than that of the nanomaterials-based or enzyme mediated amplification approaches by several orders of magnitude. More importantly, the sensor’s responsive peak current exhibits a favorable linear correlation to the logarithm of the concentrations of target sequence ranging from 1 aM to 10 pM. In addition, the sensor is highly selective, and can discriminate a single mismatched sequence. This HCR-based genosensor is also capable of probing low-abundance BRCA-1 gene sequence directly in complex matrices , such as 50% human serum, with minimal interference. These advantages will make our tailor-engineered HCR-based electrochemical genosensor appealing to genetic analysis and clinical diagnostics.
研究领域:
遗传学
·
生物医学工程
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Protocols
·
2024
·
IF 13.1
Chen Feng,Li Xinyin,Bai Min,Zhao Yongxi
Epigenetic modifications and spatial proximities of nucleic acids and proteins play important roles in regulating physiological processes and disease progression. Currently available cell imaging methods, such as fluorescence in situ hybridization (FISH) and immunofluorescence, struggle to detect low-abundance modifications and their spatial proximities. Here we describe a step-by-step protocol for three DNA-encoded amplifying FISH-based imaging strategies to overcome these challenges for varying applications: base-encoded amplifying FISH (BEA-FISH), pairwise proximity-differentiated amplifying FISH (PPDA-FISH) and cellular macromolecules-tethered DNA walking indexing (Cell-TALKING). They all use the similar core principle of DNA-encoded amplification, which transforms different nonsequence molecular features into unique DNA barcodes for in situ rolling circle amplification and FISH analysis. This involves three key reactions in fixed cell samples: target labeling, DNA encoding and rolling circle amplification imaging. Using this protocol, these three imaging strategies achieve in situ counting of low-abundance modifications alone, the pairwise proximity-differentiated visualization of two modifications and the exploration of multiple modifications around one protein (one-to-many proximity), respectively. Low-abundance modifications, including 5-hydroxymethylcytosine, 5-formylcytosine, 5-hydroxymethyluracil and 5-formyluracil, are clearly visualized in single cells. Various combinatorial patterns of nucleic acid modifications and/or histone modifications are found. The whole protocol takes ~2–4 d to complete, depending on different imaging applications.
研究领域:
分子生物学
·
细胞生物学
·
基因组学
·
表观遗传学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF FISH BIOLOGY
·
2016
·Vol. 88
·期 5
·1693-1707
·
IF 1.246
W. T. Zhang,K. Liu,J. S. Xiang,L. Y. Zhang,W. J. Liu,Z. D. Dong,Y. Z. Li,H. L. Li,S. L. Chen,N. Wang
To elucidate the effect of thyroid hormone receptor α A ( thraa ) on metamorphosis, the full length cDNA of half-smooth tongue sole Cynoglossus semilaevis was cloned. The relative gene transcript level of thraa at different development stages was quantified using real-time PCR. Transcription of thraa increased and declined rapidly during metamorphosis. Hyperthyroidism was induced in juveniles and larvae with exposure to T3 and T4, and hypothyroidism with thiourea (TU), 2-mercapto-1-methylimidazole (MMI). thraa mRNA was higher in fish treated for 6 days with MMI than in untreated controls, although inhibited larvae did not complete metamorphosis. The addition of exogenous T4 reversed this effect in the MMI-treated group, but not in the TU-treated group. In situ hybridization revealed progressive tail end of body growth and change during developmental stages, with corresponding changes in thraa expression. This process may be induced by thyroid hormones with thraa as a major mediator. The morphological changes of tip of the tail may be associated with the development of lateral swimming.
研究领域:
分子生物学
·
内分泌学
·
发育生物学
·
水产养殖
✓ 已验证
DOI
PubMed
高影响因子
EUPHYTICA
·
2024
·Vol. 220
·期 4
·1-13
·
IF 1.6
Zhong Yuan,Du Ming-jie,Ji Run-ze,Rong Qi,Guan Zi-heng,Cheng Fang-yun
Cross-incompatibility is the major challenge in inter-sectional distant hybridization between tree peonies ( Paeonia sect. Moutan ) and herbaceous peonies ( Paeonia sect. Paeonia ). Most of the intersectional cross combinations rarely produce hybrid progenies or even seeds. However, a combination with Paeonia × lemoinei Rehd. ( P. delavayi × P. suffruticosa , tree type) as the pollen parent and P. lactiflora (herbaceous) as the seed parent resulted in many progenies. This research focuses on cytogenetics of two highly compatible intersectional combinations, P. lactiflora ‘Martha W.’ × P. × lemoinei ‘Golden Era’ (‘MW’ × ‘GE’) and P. lactiflora ‘Fen Yun Fei He’ × P. × lemoinei ‘L’Esperance’ (‘FYFH’ × ‘LE’). Karyotype analysis combined with genomic in situ hybridization (GISH) showed that the seed parents, ‘MW’ and ‘FYFH’, are both diploids (2 n = 2 x = 10, CC), while the pollen parents, ‘GE’ and ‘LE’, are tetraploid (2 n = 4 x = 20, AABB) and diploid (2 n = 2 x = 10, AB) respectively. Four progenies from ‘MW’ × ‘GE’, and another four progenies from ‘FYFH’ × ‘LE’, are all triploids (2 n = 3 x = 15, ABC). P. × lemoinei contributes two sets of chromosomes (AB) showing distinct signals in all tested triploids. The essential of a successful intersectional hybridization is to obtain 2 x gametes from tree peony. P. × lemoinei has better compatibility in intersectional hybridization than other tree peony species, probably because of its higher frequency of 2 x gamete (unreduced in diploid, or reduced in tetraploid) generation. Tetraploid tree peonies, such as ‘GE’, might produce abundant 2 x gametes through normal meiosis, which is the possible reason for their better fertility and superior intersectional compatibility than diploid tree peonies. The mechanism of compatibility in intersectional hybridization is supposed to be a rebalance between the distant divergent parents, tree peonies and herbaceous peonies, triggered by ploidy increase of gametes from tree peonies. The discovery of tetraploid tree peony also provides valuable materials to study the mechanism of polyploidization in sect. Moutan , and to promote the distant hybridization breeding in Paeonia.
研究领域:
植物学
·
遗传学
·
植物育种
✓ 已验证
DOI
高影响因子
Nanoscale
·
2018
·Vol. 10
·期 23
·11079-11090
·
IF 7.233
Jingpu Zhang,Yanlei Liu,Xiao Zhi,Chunlei Zhang,Tie Fu Liu,Daxiang Cui
Dysregulation of microRNAs (miRNAs) is correlated with cancer progression. In vitro detection methods using extracts from cell lysis cannot provide information about the spatial distribution of miRNAs. Due to the development of miRNA fluorescence in situ hybridization (FISH), increasing amounts of intracellular expression information are being obtained. However, miRNA FISH suffers from weak signals and complex steps and thus remains very challenging. Herein, a strategy based on DNA-templated silver nanoclusters (AgNCs/DNAs) and their G-rich fluorescence enhancement effect was developed for FISH detection of miRNAs in gastric cancer cells. The method combines hybridization and signal amplification into one step, which allows imaging of intracellular miRNAs immediately after hybridization. Most importantly, using the method based on our design, miR-101-3p, miR-16-5p and miR-19b-3p were found to be located in the nuclei of MGC803 cells with granulated shapes, indicating an unanticipated distribution pattern. In addition, before the final miRNA FISH, we performed an optimization of AgNCs/DNAs and their G-rich fluorescence enhancement effect; we found that the effect occurred at shorter wavelengths emitting green fluorescence, with weakened red fluorescence at longer wavelengths. However, the components involved in the FISH process impacted the fluorescence properties so greatly that the probes finally exhibited slightly strengthened red fluorescence signals. Our method enables facile visualization of miRNAs at the subcellular level, which may benefit the precise localization of miRNAs in single cells in the future.
研究领域:
癌症生物学
·
分子生物学
·
纳米技术
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
NEUROPHARMACOLOGY
·
2022
·
IF 5.273
Jingli Chen,Chang Liu,Mu Xu,Jiaxi Zhu,Zhongyuan Xia
Chronic stress is associated with impairment of synapse plasticity in hippocampus and cognitive dysfunction in rodent and human. Notably, corticosterone (CORT) is believed to take responsibility for dendritic atrophy and reduction of spine number induced by chronic stress in hippocampus. But little is known about the molecular mechanisms underlying CORT induced abnormal synapse plasticity and cognitive dysfunction. Drebrin is an F-actin binding protein that modulates memory formation and maintenance by controlling the genesis and morphology of dendritic spines . In addition, miRNAs have been reported to participate in the negative regulation of protein-coding genes. In this study, five miRNAs capable of targeting Drebrin were selected by searching miRNA databases. One of these miRNAs, miR-19b-3p, was found to be upregulated in the hippocampal neurons of mice with chronic restraint stress (CRS). Luciferase reporter assay and Fluorescence in situ hybridization (FISH) were employed to identify the interaction between miR-19b-3p and Drebrin. In addition, silencing miR-19b-3p expression in vivo using an antagomir or in vitro using an inhibitor increased Drebrin expression , ameliorated the abnormal dendritic structure and upregulated the spine density in hippocampal CA1 pyramidal neurons of CRS mice and primary hippocampal neurons cultured under CORT stimulation, respectively. Electrophysiological analysis revealed that inhibition of miR-19b-3p rescued the limited synaptic transmission and synaptic plasticity in hippocampal neurons. Moreover, blocking miR-19b-3p drastically protected against cognitive deficits in CRS mice. These in vivo and in vitro findings indicate that the upregulation of miR-19b-3p exacerbates CRS-induced abnormal synaptic plasticity and cognitive impairment by targeting Drebrin.
研究领域:
神经科学
·
分子生物学
·
认知科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMOSPHERE
·
2022
·
IF 8.943
Xiaoqi Dong,Zidan Huang,Xingxing Peng,Xiaoshan Jia
A novel coupled yeast-sludge system (CYSS) was constructed by the yeast Candida sp. PNY integrated with activated sludge to treat non-sterile mainstream wastewater. After 240-day cultivation, compared with single activated sludge , simultaneous removal efficiency of total organic carbon (TOC), nitrogen and phosphorus increased by 19.5% (176.34 mg TOC g −1 d −1 ), 21.3% (11.25 mg TN g −1 d −1 ) and 15.0% (6.95 mg TP g −1 d −1 ), respectively, while the amount of sludge reduced by 50%. Amplicon sequencing analysis showed that the abundance of Nitrosomonas , Nitrospira , Zoogloea , Dechloromonas , and Candidatus Accumulibacter significantly decreased to 0% on Day 200. Abundance of nirS and nirK for denitrification significantly decreased in CYSS by quantitative PCR (qPCR), and the copies of nirS and nirK were 3.37-fold and 1.71-fold decrease from Day 0 to Day 240, respectively. The results of Fluorescence in situ hybridization and co-occurrence network showed that Candida sp. PNY predominated its distribution in CYSS, and strongly connected with environmental variables based on network analysis. Furthermore, this study reconstructed the carbon, nitrogen and phosphorus metabolic pathways of the CYSS based on metagenomics .
研究领域:
环境工程
·
微生物学
·
废水处理
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2025
·Vol. 15
·期 1
·1-11
·
IF 3.9
Yu Xiang-Fang,Gurung Chetali,Yu Zhongjia,Xiao Wei,Kerem Goher,Teng Bin,Zhang Jian V.,Su Zhe,Zhou Yongjin,Ren Pei-Gen
Awareness of the intestinal microflora involved in insulin resistance and type 2 diabetes (T2DM) has now become more evident. However, direct mechanical insight is required to illustrate the contribution of intestinal microflora in the disease progression. In this study, we aimed to precisely assess the changes in intestinal bacteria translocation (IBT) from gut to the pancreas using combinations of FISH, 16S rRNA amplicon sequencing, and deep learning-assisted methods to track IBT in diet-induced obese (DIO) and antibiotic-induced microbiota disruption (AIMD)-DIO mouse models. Our analysis showed deep learning-assisted quantification enhanced the accuracy and objectivity of bacterial tracking. The DIO mice exhibited increased IBT, likely due to excessive intestinal lipid accumulation and compromised intestinal barrier integrity. Elevated bacterial loads in the pancreas were associated with worsened pancreatic function, indicated by higher fasting blood glucose, impaired glucose tolerance, and dysfunctional insulin secretion. In contrast, the AIMD-DIO group lowered the IBT, maintained the islet structure and improved glucose homeostasis. Comparative study between DIO and AIMD-DIO models revealed a strong correlation between number of translocated bacteria and T2DM severity. These findings provide objective evidence of bacterial migration from the intestine to the pancreas and establishes its pathological relationship with pancreatic impairment and dysfunction and underscores to utilize AI techniques more successfully in the future for evaluating IBT.
研究领域:
微生物学
·
糖尿病研究
·
分子生物学
·
人工智能
·
免疫学
·
代谢性疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
International Journal of Nanomedicine
·
2024
·
IF 6.6
Yi He,Lejia Zhang,Siyuan Huang,Yuquan Tang,Yiming Li,Hongyu Li,Guanhui Chen,Xun Chen,Xiliu Zhang,Wei Zhao,Feilong Deng,Dongsheng Yu
Background The ability of nanomaterials to induce osteogenic differentiation is limited, which seriously imped the repair of craniomaxillofacial bone defect. Magnetic graphene oxide (MGO) nanocomposites with the excellent physicochemical properties have great potential in bone tissue engineering. In this study, we aim to explore the craniomaxillofacial bone defect repairment effect of MGO nanocomposites and its underlying mechanism.Methods The biocompatibility of MGO nanocomposites was verified by CCK8, live/dead staining and cytoskeleton staining. The function of MGO nanocomposites induced osteogenic differentiation of BMSCs was investigated by ALP activity detection, mineralized nodules staining, detection of osteogenic genes and proteins, and immune-histochemical staining. BMSCs with or without MGO osteogenic differentiation induction were collected and subjected to high-throughput circular ribonucleic acids (circRNAs) sequencing, and then crucial circRNA circAars was screened and identified. Bioinformatics analysis, Dual-luciferase reporter assay, RNA binding protein immunoprecipitation (RIP), fluorescence in situ hybridization (FISH) and osteogenic-related examinations were used to further explore the ability of circAars to participate in MGO nanocomposites regulation of osteogenic differentiation of BMSCs and its potential mechanism. Furthermore, critical-sized calvarial defects were constructed and were performed to verify the osteogenic differentiation induction effects and its potential mechanism induced by MGO nanocomposites.Results We verify the good biocompatibility and osteogenic differentiation improvement effects of BMSCs mediated by MGO nanocomposites. Furthermore, a new circRNA-circAars, we find and identify, is obviously upregulated in BMSCs mediated by MGO nanocomposites. Silencing circAars could significantly decrease the osteogenic ability of MGO nanocomposites. The underlying mechanism involved circAars sponging miR-128-3p to regulate the expression of SMAD5, which played an important role in the repair craniomaxillofacial bone defects mediated by MGO nanocomposites.Conclusion We found that MGO nanocomposites regulated osteogenic differentiation of BMSCs via the circAars/miR-128-3p/SMAD5 pathway, which provided a feasible and effective strategy for the treatment of craniomaxillofacial bone defects.
研究领域:
生物材料
·
组织工程
·
骨再生
·
分子生物学
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
Frontiers in Genetics
·
2021
·
IF 4.599
Li Jia,Shen Jie,Qin Lan,Lu Dongyan,Ding Enci
Background: Thyroid cancer is a frequent endocrine tumor in women. It is of great significance to investigate the molecular mechanism of progression of thyroid cancer.Methods: Gene expression data set and clinical data were downloaded from The Cancer Genome Atlas database for differential expression analysis. The triplet of downstream transcription factors (TFs) and modulatory genes of target lncRNA in thyroid cancer was predicted by the lncMAP database. mRNA and protein expression of lncRNA LBX2-AS1, RARα, and FSTL3 were detected by qRT-PCR and western blot. The localization of lncRNA LBX2-AS1 in cells was tested by Fluorescence in situ hybridization assay. The RNA immunoprecipitation assay was applied to verify the binding relationship between lncRNA LBX2-AS1 and FSTL3. ChIP and dual-luciferase assays were used to prove the binding relationship between RARα and FSTL3. Cell function experiments were used to test cell proliferation, migration and invasion in each treatment group. The role of lncRNA LBX2-AS1 in thyroid cancer progression was also confirmed in nude mice.Results: Bioinformatics analysis indicated that lncRNA LBX2-AS1, RARα, FSTL3 were remarkably fostered in thyroid cancer tissue, and LBX2-AS1 was evidently correlated with clinical features. The LncMAP triplet prediction showed that LBX2-AS1 recruited TF RARα to modulate FSTL3. RIP assay confirmed that LBX2-AS1 was prominently enriched on RARα. ChIP and dual-luciferase report assays unveiled that RARα bound to the promoter region of FSTL3 and functioned as a TF. Cell function experiments uncovered that LBX2-AS1 boosted the progression of thyroid cancer. The rescue experiments showed that LBX2-AS1 recruited the TF RARα to hasten the transcription activity of FSTL3 and thus promoted the development of thyroid cancer.Conclusion: The integrative results demonstrated that LBX2-AS1 activated FSTL3 by binding to TF RARα to hasten proliferation, migration and invasion of thyroid cancer.
研究领域:
肿瘤学
·
分子生物学
·
癌症遗传学
·
生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）