Journal: Insects

Article Title: MicroRNA-281-X Modulates Self-Grooming Behavior in Honeybees by Targeting Tyrosine Decarboxylase 2 in the Octopaminergic Pathway

doi: 10.3390/insects17050522

Figure Lengend Snippet: Tissue-specific expression profile and interactions between miR-281-x and its target tdc2 in honeybees. ( A ) Relative expression levels of miR-281-x and tdc2 from eclosion to 21-day-old adult bees. The smooth curve was fitted using the LOWESS spline method to visualize the dynamic expression of miR-281-x and tdc2 ( n = 3 per group). ( B ) Distribution of miR-281-x and tdc2 gene expression in different tissues of honeybees ( n = 3 per group, Kruskal–Wallis test). Different lowercase letters (a, b, c) represent significant differences ( p < 0.05); ns denotes not significant ( p ≥ 0.05). ( C ) Dual luciferase reporter assays in wild and mutant types of tdc2 in vitro ( n = 6 per group). Statistical comparisons were performed using the Kruskal–Wallis test, followed by post hoc pairwise comparisons. Effect sizes (r) for pairwise comparisons. ** p < 0.01. ( D ) RNA immunoprecipitation (RIP) was performed with an anti-Ago-1 antibody, and fold enrichment was quantified relative to rabbit IgG control. qPCR analysis was performed to amplify tdc2 mRNA from the Ago-1 immunoprecipitates in brain tissue extracts treated with agomir-281-x compared with agomir-NC. Significant differences were determined by the Student’s t -test ( n = 3 per group, * p < 0.05; ** p < 0.01). ( E ) miR-281-x and tdc2 were co-labeled to determine the co-localization in the honeybee brain by fluorescence in situ hybridization (FISH). Where green ( tdc2 ) and red signals (miR-281-x) overlap, a yellow signal is observed, indicating the co-localization of miR-281-x and tdc2 . The images were visualized using a laser confocal microscope (Zeiss) at magnifications of ×12 (the left column) and ×40 (the right column), scale bars: 200 µm (left column) and 50 µm (right column).

Article Snippet: Briefly, digoxigenin-labeled miR-281-x and tdc2 DNA probes were designed using miRBase and NCBI BLASTN (performed by Servicebio, China) and synthesized by Servicebio.

Techniques: Expressing, Gene Expression, Luciferase, Mutagenesis, In Vitro, RNA Immunoprecipitation, Control, Labeling, Fluorescence, In Situ Hybridization, Microscopy

Journal: Insects

Article Title: MicroRNA-281-X Modulates Self-Grooming Behavior in Honeybees by Targeting Tyrosine Decarboxylase 2 in the Octopaminergic Pathway

doi: 10.3390/insects17050522

Figure Lengend Snippet: miR-281-x negatively regulates the expression of tdc2 . ( A ) Relative mRNA level ( n = 5 per group) and protein level ( n = 5 per group) of tdc2 in the brains of bees injected with agomir-281-x. ( B ) Relative mRNA level ( n = 5 per group) and protein level ( n = 5 per group) of tdc2 in the brains of bees injected with antagomir-281-x. ( C ) Relative mRNA level ( n = 6 per group) and protein level ( n = 5 per group) of tdc2 in the brains of MS and MW bees. The data are presented as the mean ± SEM; The asterisks outside the strip indicate the significant difference between controls and the treatments by Student’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001. Protein: Tdc2: 72 kDa; GAPDH: 36 kDa.

Article Snippet: Briefly, digoxigenin-labeled miR-281-x and tdc2 DNA probes were designed using miRBase and NCBI BLASTN (performed by Servicebio, China) and synthesized by Servicebio.

Techniques: Expressing, Injection, Stripping Membranes

Journal: Insects

Article Title: MicroRNA-281-X Modulates Self-Grooming Behavior in Honeybees by Targeting Tyrosine Decarboxylase 2 in the Octopaminergic Pathway

doi: 10.3390/insects17050522

Figure Lengend Snippet: miR-281-x controls octopamine signaling by targeting tdc2 modulates self-grooming behavior. ( A ) First grooming time (s) and total grooming bouts after injecting dstdc2 . Group differences among dsEGFP ( n = 88) and dstdc2 ( n = 83) bees were tested by the Mann–Whitney U test, ** p < 0.01. ( B ) Behavioral rescue assay was performed by injecting sitdc2 into honeybees pretreated with antagomir-281-x (the Mann–Whitney U test * p < 0.05, antagomir-281-x + siNC : n = 60; antagomir-281-x + sitdc2 : n = 49). ( C , D ) Concentrations of octopamine in the brains of bees ( n = 18 per group) injected with agomir-281-x and antagomir-281-x. ( E ) Concentrations of octopamine in the brains of bees ( n = 20 per group) with tdc2 RNAi knockdown. The data are presented as the mean ± SEM; The asterisks outside the strip indicate the significant difference between controls and the treatments by Student’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Briefly, digoxigenin-labeled miR-281-x and tdc2 DNA probes were designed using miRBase and NCBI BLASTN (performed by Servicebio, China) and synthesized by Servicebio.

Techniques: MANN-WHITNEY, Rescue Assay, Injection, Knockdown, Stripping Membranes

Journal: Insects

Article Title: MicroRNA-281-X Modulates Self-Grooming Behavior in Honeybees by Targeting Tyrosine Decarboxylase 2 in the Octopaminergic Pathway

doi: 10.3390/insects17050522

Figure Lengend Snippet: The miR-281-x– tdc2 –octopamine axis regulates self-grooming in A. mellifera . miR-281-x acts as a negative regulator by directly targeting tdc2 , a key enzyme for octopamine synthesis. High miR-281-x levels inhibit tdc2 translation and reduce octopamine, resulting in weak grooming. Conversely, low miR-281-x levels facilitate tdc2 expression and octopamine accumulation, driving intense self-grooming behavior.

Article Snippet: Briefly, digoxigenin-labeled miR-281-x and tdc2 DNA probes were designed using miRBase and NCBI BLASTN (performed by Servicebio, China) and synthesized by Servicebio.

Techniques: Expressing

Journal: RSC Advances

Article Title: Graphene field-effect transistor based multiplexed sensing platform for simultaneous detection of multiple Alzheimer's disease biomarkers

doi: 10.1039/d5ra07384g

Figure Lengend Snippet: The electrical sensing results of GFET biosensor. (a) (PBASE: 5 mM; aptamer: 5 µM). (b) GFET transfer curves for Aβ42 detection under identical functionalization. (c) Four regions functionalized with Aβ42 and hsa-miR-125b aptamers in 1× PBS, with Aβ42 target added for measuring the average signal response at different concentrations. (d) Four regions functionalized with Aβ42 and hsa-miR-125b aptamers in 1× PBS, with hsa-miR-125b target added for measuring the average signal response at different concentrations. (e) The sensor response as a function 125b concentration, with mean, standard deviation, median, and a linear fitting curve. (f) The sensor response as a function Aβ42 concentration, with mean, standard deviation, median, and a linear fitting curve ( n ≥ 13, n is number of sensors used for each measurement) ( n ≥ 13, n is number of sensors used for each measurement).

Article Snippet: Aβ42 and Aβ40 peptides, hsa-miR-125b RNA sequence, the hsa-miR-125b complementary DNA probe and the Aβ42 DNA aptamer were synthesized by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Concentration Assay, Standard Deviation

Journal: RSC Advances

Article Title: Graphene field-effect transistor based multiplexed sensing platform for simultaneous detection of multiple Alzheimer's disease biomarkers

doi: 10.1039/d5ra07384g

Figure Lengend Snippet: Mixed sample detection results. (a) Higher concentration of Aβ42 biomarker vs. lower concentration of hsa-miR-125b biomarker. The absolute concentrations were (Aβ42 : 125b): 10 pM : 1 fM (10 000 : 1); 50 nM : 50 pM (1000 : 1); and 100 nM : 1 nM (100 : 1). (b) Higher concentration of hsa-miR-125b biomarker vs. lower concentration of Aβ42 biomarker. The absolute concentrations were (125b : Aβ42): 100 nM : 1 fM (10 8 : 1), 100 nM : 1 pM (10 6 : 1), and 50 nM : 50 pM (10 3 : 1). (c) Fixed concentration of Aβ40 biomarker and varying concentrations of hsa-miR-125b ( n ≥ 13, n is number of sensors used for each measurement).

Article Snippet: Aβ42 and Aβ40 peptides, hsa-miR-125b RNA sequence, the hsa-miR-125b complementary DNA probe and the Aβ42 DNA aptamer were synthesized by Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Concentration Assay, Biomarker Discovery

Journal: bioRxiv

Article Title: Chromosomal variability in a clonal crop: Somaclonal change follows the emergence of triploid saffron crocus

doi: 10.64898/2026.05.04.722608

Figure Lengend Snippet: Fluorescent in situ hybridisation (FISH) of mitotic chromosomes of four different C. sativus lineages from Spain, Egypt, Germany, and Kashmir (lineage numbers are shown on the panel). DAPI-stained chromosomes are shown in gray. FISH of different C. sativus accessions (A); Close up of chromosomes 1.1-1.3 and chromosome 5.1 (SatDNA names given in each panel) showing the number of signals for each satDNA (B); FISH karyotype of C. sativus (C). Probes used are CroSat1 (blue), CroSat2 (green), CroSat3 (aqua), CroSat4 (red), 5S rRNA genes (white) and 18S-5.8S-25S rRNA genes (yellow). Arrows indicate extra signals found compared to the reference accession. Bar is 10 µm.

Article Snippet: Satellite DNA probes (CroSat1-4; ) were labeled by PCR with fluorescently labelled UTP, DY415-dUTP, 495-dUTP, 547-dUTP, and 647-dUTP (Dyomics GmbH, Jena, Germany).

Techniques: In Situ, Hybridization, Staining

Journal: Horticulture Research

Article Title: PtrbZIP12 improves drought resistance in Populus trichocarpa by directly targeting PtrDHN and PtrPOD

doi: 10.1093/hr/uhag034

Figure Lengend Snippet: PtrbZIP12 directly activates the promoters of PtrDHN and PtrPOD . (a) Relative expression levels of PtrDHN and PtrPOD in PtrbZIP12 -OE versus WT plants. Data are presented as means ± SD from three biological replicates. (b) Verification of the direct regulatory interaction between PtrbZIP12 and the PtrDHN promoter using ChIP-PCR, in which the promoter region (−800 to −1 bp) was divided into four fragments (P1–P4). ‘Input’ represents chromatin before immunoprecipitation, ‘ChIP-’ indicates immunoprecipitation without antibody, and ‘ChIP+’ denotes immunoprecipitation with anti-GFP antibody. (c) Y1H assay demonstrating PtrbZIP12 binding to ABRE motifs; controls included p53-HIS2/pGADT7-Rec2-p53 (positive) and pGADT7-Rec2- PtrbZIP12 /p53-HIS2 (negative). (d and e) Y1H assays showing PtrbZIP12 binding to the promoters of PtrDHN and PtrPOD , with P53-promoter-AUR1-C and AD-Rec-P53 as positive controls, and AD-empty prey vector with AUR1-C driven by the target gene promoter as negative controls. (f) EMSA confirming PtrbZIP12 –ABRE binding; lanes: 1, biotin-labeled probe; 2, labeled probe + PtrbZIP12 protein; 3–5, competition with 10-, 50-, and 100-fold molar excess of unlabeled probe. (g and h) Schematic representation of effector and reporter constructs utilized in the dual-LUC assay, with transient LUC/renillase (REN) coactivation experiments in N. benthamiana leaves demonstrating PtrbZIP12 -mediated activation of PtrDHN (g) and PtrPOD (h).

Article Snippet: Specific biotin-labeled DNA probes targeting the promoter region of the gene were synthesized by Sangon Biotech Co.

Techniques: Expressing, Immunoprecipitation, Y1H Assay, Binding Assay, Plasmid Preparation, Labeling, Construct, Activation Assay