Journal: Fundamental Research

Article Title: Single-cell multi-omics analysis decodes molecular characteristics of sheep oocyte fate in vivo maturation

doi: 10.1016/j.fmre.2025.10.009

Figure Lengend Snippet: Modulating sheep oocyte developmental fate through ET-1 supplement. (a) The rate of polar body extrusion after 18 h IVM in ET-1, DMSO, BQ123. (b) The fluorescence intensity of ROS signals was compared in ET-1, DMSO, and BQ123. (c) Representative images of ROS levels of mature oocytes in the ET-1, DMSO, BQ123. Scale bar, 50 µm. Representative images of GSH levels of mature oocytes in the ET-1, DMSO, BQ123. Scale bar, 100 µm. (d) The fluorescence intensity of GSH signals was compared in ET-1, DMSO, and BQ123. (e) Representative images of Mitochondrial membrane potential (ΔΨm), detected by JC-1 staining in ET-1, DMSO, BQ123. (f) The ratio of red to green fluorescence intensity indicates the mitochondrial membrane potential level in the ET-1, DMSO, BQ123. (g) Representative images of the mitochondrial distribution of mature oocytes in the ET-1, DMSO, BQ123, detected by Mito-Tracker Red. Scale bar = 50 µm. (h) The rate of abnormal mitochondrial distribution in ET-1, DMSO, BQ123. (i) Representative images of the CGs distribution of an equatorial section of mature oocytes in the ET-1, DMSO, BQ123, detected by FITC-PNA. Scale bar = 50 µm. (j) The rate of normal CGs distribution in ET-1, DMSO, BQ123. (k) Representative images of the spindle morphology and chromosome alignment of mature oocytes in the ET-1, DMSO, BQ123, detected by immunofluorescent staining. Scale bar = 5 µm. (l) The rate of normal Spindle/chromosome distribution in ET-1, DMSO, BQ123. (m) Representative images of the blastocyst after 7.5 dpi IVF of ET-1, DMSO, BQ123. (n) The rate of blastocyst formation at 7.5 dpi was compared in ET-1, DMSO, and BQ123. (o) Representative images of immunofluorescent staining of CDX2 and OCT4 in blastocysts in ET-1, DMSO, BQ123, with DAPI for DNA. Scale bar = 50 µm. (p) The ratio of Blastocysts’ ICM/TE was compared in ET-1, DMSO, and BQ123. Letters that are the same indicate no significant difference ( P > 0.05), while different letters indicate significant differences ( P ≤ 0.05).

Article Snippet: To screen for the optimal working concentration, ET-1 (Sigma, 05-23-3800) and BQ123 (MCE, HY-12378) were supplemented in the IVM medium at concentrations of 1 nM, 10 nM, and 100 nM, respectively.

Techniques: Fluorescence, Membrane, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Tumor-targeted IL2 promotes specific CD8 + T cells private clonal expansion enhancing lymphoma control

doi: 10.1186/s13046-026-03678-7

Figure Lengend Snippet: A Schematic representation of the 2-photon intravital microscopy (2P-IVM). B Plots of cell tracks with common origin showing longer tracks in L19IL2 treated CD8 + T cells in comparison to the same field of view before L19IL2 administration. Cell motility quantification using track maximum ( C ) speed, ( D ) mean speed, ( E ) length, ( F ) displacement, ( G ) speed variation, and ( H ) straightness. I CD8 + T cell tracks in untreated (up) and at 30 min post treatment (down) relative to lymphoma cells (red). J Pixel motility heatmap in untreated (up) and treated mice (down) showing hotspots with high motility (yellow). K right Equidistant regions of interest (ROIs) showing the different tumor regions from peripheral (I.) to deep areas (IV.), applied to pixel motility heatmap (left; up untreated and down L19IL2 treated). L Quantification of pixel velocity for each concentric ROI before (red) and after (green) L19IL2 administration. In ( C - H ) circles represent individual track values of all CD8. + T cells from one representative mouse. In all graphs, the p -value is indicated as * < 0.05; ** < 0.01; *** < 0.001; **** < 0.0001

Article Snippet: Image Analysis and Data Processing Cell detection, tracking and volumetric reconstruction from 4D 2P-IVM data were performed using Imaris (Oxford Instruments, v10.2.0).

Techniques: Intravital Microscopy, Comparison

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) For in vivo maturation (yellow box), 4 to 6-week-old female C57Bl/6J mice were stimulated with pregnant mare serum gonadotropin (PMSG) for 46 hours, triggered with human chorionic gonadotropin (hCG) and mature cumulus-oocyte complexes (COCs) were retrieved 14 hours post-trigger. For in vitro culture groups, 4 to 6-week-old female C57Bl/6J mice were stimulated with PMSG and ovaries collected 23 hours later to retrieve immature COCs. For the CAPA treatment group (pink box) COCs were cultured in CAPA pre-medium for 24 hours and then in vitro maturation (IVM) medium containing listed additives for 18 hours. For the standard IVM treatment group (blue box), COCs were cultured in IVM medium for 18 hours. COCs were then used intact for embryology assessments or separated into oocytes and cumulus cells for proteomics and validation experiments. B) For protein assessment in human oocytes, patients underwent a full stimulation protocol for an intracytoplasmic sperm injection (ICSI) cycle as per standard clinical procedures. Following oocyte pickup, immature germinal vesicle (GV) oocytes were collected from the clinic and transported to the lab. Oocytes were allocated into two groups, either fixed at the GV stage, or oocytes underwent rescue-IVM for 24hrs to reach the metaphase II (MII) stage. Immunostaining experiments were performed on both cohorts of oocytes.

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: In Vivo, In Vitro, Cell Culture, Biomarker Discovery, Injection, Immunostaining

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) Brightfield images of cumulus-oocyte complexes (COCs) following in vivo maturation (yellow), CAPA IVM (pink) and standard IVM (IVM; blue). Red circles mark oocytes, and scale bar is 50 µm. Diagram of in vitro fertilisation (IVF) followed by morphological assessment of embryos at day 2, 5 and 6 of in vitro embryo culture. B) Fertilisation rate (percentage of cleaved embryos per oocyte transferred to culture), D) Day 5 blastocyst rate, F) Day 6 blastocyst rate and G) Day 6 hatching blastocyst rate (percentage of hatching blastocysts per cleaved embryo) following oocyte maturation. Corresponding images of embryos at Day 2 (C), Day 5 (E) and Day 6 (H) are shown. Red arrowheads mark cleaved embryos, red circles show oocytes that failed to cleave, hatched blastocysts (red h), blastocyst (red bl) and fragmented embryos (red f). Images taken at 10x magnification and scale bar denotes 100 µm. All embryo outcome data were arcsine transformed and one-way ANOVA followed by Tukey’s post hoc tests were performed. Numbers above the x axis denote the total number of oocytes (panel B) and total number of cleaved embryos (panel D, F, G) in each oocyte group across the six replicates. Bars with no common superscripts are significantly different (P < 0.05).

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: In Vivo, In Vitro, Embryo Culture, Transformation Assay

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) The total number of proteins identified, average number of unique peptides identified/protein and the average protein coverage (%) within oocytes matured in vivo (yellow), following CAPA-IVM (pink) and following standard IVM (blue). B) The total number of proteins identified, average number of unique peptides identified/protein and the average protein coverage (%) within cumulus cells matured in vivo (light yellow), following CAPA-IVM (light pink) and following standard IVM (light blue). Principal component analysis (PCA) from each maturation method ( in vivo (yellow squares), CAPA-IVM (pink circles) and standard IVM (blue diamonds)) for both C) oocytes and D) cumulus cells. E) Venn diagram of shared and unique proteins across the three groups of oocytes. F) Venn diagram of shared and unique proteins across the three groups of cumulus cells.

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: In Vivo

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) Volcano plots showing pairwise comparisons of differential oocyte protein expression in in vivo (yellow), CAPA-IVM (pink) and standard IVM (blue) treatment groups. Coloured dots indicate differentially expressed proteins in each comparison and grey dots indicate proteins below the log2FC ± 1, p-value ≤ 0.05 cutoff. B) All proteins found to be consistently differentially expressed with log2 fold-change and −log10 p- values of CAPA-IVM matured MII oocytes and standard IVM matured MII oocytes compared with in vivo matured MII oocytes (IVO). Bar charts showing the top 20 enriched canonical pathways based on oocyte differential protein expression between C) CAPA- IVM matured MII oocytes and in vivo matured MII oocytes, and D) standard IVM matured MII oocytes and in vivo matured MII oocytes. Shades of green indicate -log10 p value generated from Ingenuity Pathway Analysis software. Scaled proteomic abundances for individual proteins related to biological pathway elevated in E) in vivo oocytes and F) in vitro matured oocytes (n = 4, abundance ratio adjusted p-values and mean ± standard deviation).

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: Expressing, In Vivo, Comparison, Generated, Software, In Vitro, Standard Deviation

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) Eukaryotic initiation factor 2A (EIF2A, magenta), B) ribosomal protein L24 (RPL24, magenta) and C) clathrin light chain B (CLTB, magenta) immunocytochemistry in MII oocytes following in vivo maturation (yellow), CAPA IVM (pink) and standard IVM (IVM; blue). Nuclei were counterstained with Hoechst (blue). Images taken at 40x magnification and scale bar denotes 25 µm. Fluorescence intensity of (D) EIF2A, (E) RPL24 and (F) CLTB was quantified in oocytes from MII COCs following in vivo maturation (yellow), CAPA IVM (pink) and standard IVM (IVM; blue). Integrated density values were taken from individual COCs and normalised to the in vivo control mean intensity of each biological replicate (dots). Data are presented as the mean of three replicates ± SEM. Statistical analyses were performed using one-way ANOVA followed by Tukey’s post hoc tests. Bars with no common superscripts are significantly different (P < 0.05). A.U, arbitrary units. G) Immunofluorescent staining for translation machinery (RPL24 and EIF2A) and endocytosis (CLTB) in human prophase I (GV) and metaphase II oocytes. Nuclei were counterstained with Hoechst (blue). Images taken at 40x magnification and scale bar denotes 50 µm.

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: Immunocytochemistry, In Vivo, Fluorescence, Control, Staining

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) Volcano plots showing pairwise comparisons of differential cumulus cell protein expression in in vivo (yellow), CAPA-IVM (pink) and IVM (blue) treatment groups. Coloured dots indicate differentially expressed proteins in each comparison and grey dots indicate proteins below the log2FC ± 1, p-value ≤ 0.05 cutoff. B) All proteins found to be consistently differentially expressed with log2 fold-change and −log10 p-values of cumulus cells from CAPA-IVM and IVM maturation groups, compared with in vivo matured cumulus cells (IVO).

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: Expressing, In Vivo, Comparison

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: Bar charts showing the top 20 enriched canonical pathways based on cumulus cell differential protein expression between A) CAPA-IVM matured MII COCs and in vivo matured MII COCs, and B) IVM matured MII COCs and in vivo matured MII COCs. Shades of green indicate -log10 p value generated from Ingenuity Pathway Analysis software. Scaled proteomic abundances for individual proteins related to biological pathway elevated in C) in vivo cumulus cells and D) in vitro matured cumulus cells (n = 4, abundance ratio adjusted p-values and mean ± standard deviation).

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: Expressing, In Vivo, Generated, Software, In Vitro, Standard Deviation

Journal: bioRxiv

Article Title: A proteomic signature of oocyte quality from models of varying oocyte developmental competence

doi: 10.64898/2026.01.20.700687

Figure Lengend Snippet: A) Ferredoxin 1 (FDX1, magenta) and C) D-3-phosphoglycerate dehydrogenase (PHGDH, magenta) immunocytochemistry colocalised with phalloidin (green) in MII COCs following in vivo maturation (yellow), CAPA IVM (pink) and standard IVM (IVM; blue). Nuclei were counterstained with Hoechst (blue). First column shows intact cumulus oocyte complexes, with the subsequent columns showing representative images of cumulus cells. Images taken at 40x magnification and scale bar denotes 50 µm. Fluorescence intensity of (B) FDX1 and (D) PHGDH was quantified in cumulus cells from MII COCs following in vivo maturation (yellow), CAPA IVM (pink) and standard IVM (IVM; blue). Integrated density values were taken from individual COCs and normalised to the in vivo control mean intensity of each biological replicate (dots). Data are presented as the mean of three replicates ± SEM. Statistical analyses were performed using one-way ANOVA followed by Tukey’s post hoc tests. Bars with no common superscripts are significantly different (P < 0.05). A.U, arbitrary units.

Article Snippet: Following CAPA pre-IVM, COCs were washed 3 times in CAPA IVM media (bicarbonate-buffered αMEM + 3mg/mL fatty-acid free BSA supplemented with 50 μg/mL gentamycin, 2.5 mIU/mL FSH, 50 ng/mL recombinant mouse amphiregulin (mAREG; catalogue #989-AR, R&D Systems) and 50 ng/mL recombinant epiregulin (mEREG; catalogue #1068-ER, R&D Systems)).

Techniques: Immunocytochemistry, In Vivo, Fluorescence, Control