Journal: iScience

Article Title: Domain-autonomous MKLP1 modules divergently regulate mitotic fidelity and polyploidy in a holocentric system

doi: 10.1016/j.isci.2026.117149

Figure Lengend Snippet:

Article Snippet: rabbit anti-β-tubulin , Proteintech, Wuhan, China , Cat#10094-1-AP;RRID:AB_2210695.

Techniques: Transfection, Cloning, Imaging, Bicinchoninic Acid Protein Assay, Western Blot, Plasmid Preparation, Recombinant, Software

Journal: Advanced Science

Article Title: Gonadotropin‐Inhibitory Hormone (GnIH) and Its Receptor GPR147 Mediates Acute Stress‐Induced Activation of the Locus Coeruleus‐SAM Axis

doi: 10.1002/advs.202518996

Figure Lengend Snippet: Primer sequences for quantitative real‐time PCR.

Article Snippet: The following primary antibodies were used: β‐tubulin (Cat. 66240‐1‐Ig, proteintech, 1:2000 dilution; immunogen: recombinant human Tubulin‐beta protein; species reactivity: mouse), TH (Cat. sc‐25269, Santa Cruz, 1:100 dilution; immunogen: human TH amino acids 1–196; species reactivity: mouse), norepinephrine transporter (NET) (Cat. 680447, Zenbio, 1:1000 dilution; immunogen: a synthetic peptide of human NET; species reactivity: mouse), synaptosomal‐associated protein, 25 kDa (SNAP25) (Cat. R27294 , Zenbio, 1:1000 dilution; immunogen: human SNAP25; species reactivity: mouse), Syntaxin 1A (Cat. R389318, Zenbio, 1:1000 dilution; immunogen: human STX1A; species reactivity: mouse) and Synaptophysin (Cat. R380890, Zenbio, 1:1000 dilution; immunogen: human Synaptophysin amino acids 10–310; species reactivity: mouse).

Techniques: Real-time Polymerase Chain Reaction

Journal: Advanced Science

Article Title: Gonadotropin‐Inhibitory Hormone (GnIH) and Its Receptor GPR147 Mediates Acute Stress‐Induced Activation of the Locus Coeruleus‐SAM Axis

doi: 10.1002/advs.202518996

Figure Lengend Snippet: Activation of the GnIH/GPR147 System in the LC‐SAM Axis across Acute Restraint Stress Paradigms. (A to D): the concentration of CORT (A), NE (B), EPI (C), and GnIH (D) in the serum of mice under restraint stress (RS). (E and F): (E) The representative immunohistochemistry images and (F) mean densitometric values of GnIH or GPR147 in the locus coeruleus in acute restraint stress paradigms. (G and H): GnIH , GPR147 , TH mRNA (G) and TH protein (H) levels in the locus coeruleus across acute restraint stress paradigms. (I and J): (I) Representative immunohistochemistry images and (J) mean densitometric values of GnIH or GPR147 in the adrenal gland in acute restraint stress paradigms. (K and L): GnIH , GPR147 , TH , PNMT mRNA (K) and TH protein (L) in the locus coeruleus across diverse acute stress paradigms, including RS. (M) Optogenetic experiments demonstrated the expression of TH and GnIH neurons in the LC after the activation of norepinephrine neurons. Scale bar, 200 µm. (N and O) the concentration of GnIH (N) and NE (O) in the serum of mice after the activation of norepinephrine neurons. CORT: corticosterone; NE: norepinephrine; EPI: epinephrine; LC: locus coeruleus; TH: tyrosine hydroxylase; PNMT: phenylethanolamine N‐methyltransferase. GAPDH and β‐tubulin was used as an internal control. Data are mean ± SEM (n = 10 biological replicates per group). Comparisons between two groups were performed using two‐sided unpaired Student's t ‐test with α = 0.05. For multiple comparisons, one‐way ANOVA followed by Tukey's post‐hoc test was used. Effect sizes (Cohen's d) were calculated for all significant comparisons. * p < 0.05, ** p < 0.01, compared to the control group.

Article Snippet: The following primary antibodies were used: β‐tubulin (Cat. 66240‐1‐Ig, proteintech, 1:2000 dilution; immunogen: recombinant human Tubulin‐beta protein; species reactivity: mouse), TH (Cat. sc‐25269, Santa Cruz, 1:100 dilution; immunogen: human TH amino acids 1–196; species reactivity: mouse), norepinephrine transporter (NET) (Cat. 680447, Zenbio, 1:1000 dilution; immunogen: a synthetic peptide of human NET; species reactivity: mouse), synaptosomal‐associated protein, 25 kDa (SNAP25) (Cat. R27294 , Zenbio, 1:1000 dilution; immunogen: human SNAP25; species reactivity: mouse), Syntaxin 1A (Cat. R389318, Zenbio, 1:1000 dilution; immunogen: human STX1A; species reactivity: mouse) and Synaptophysin (Cat. R380890, Zenbio, 1:1000 dilution; immunogen: human Synaptophysin amino acids 10–310; species reactivity: mouse).

Techniques: Activation Assay, Concentration Assay, Immunohistochemistry, Expressing, Control

Journal: Advanced Science

Article Title: Gonadotropin‐Inhibitory Hormone (GnIH) and Its Receptor GPR147 Mediates Acute Stress‐Induced Activation of the Locus Coeruleus‐SAM Axis

doi: 10.1002/advs.202518996

Figure Lengend Snippet: Effects of GnIH Knockout on Anxiety‐like Behaviors and LC‐SAM Axis Activation in Mice under Acute Restraint Stress. (A to E) Effects of GnIH knockout on anxiety‐like behaviors during the open field test following acute restraint stress: (A) Motion track and heat map; (B) Number of rearing; (C) Total locomotion; (D) Distance travelled in the zone; (E) Anxiety index. (F to M) Effects of GnIH knockout on anxiety‐like behaviors during the elevated plus‐maze test following acute restraint stress: (F) Motion track and heat map; (G) Number of rearing; (H) Number of grooming; (I) Percentage of time in closed arm; (J) Percentage of entries in closed arm; (K) Total locomotion; (L) Number of entries to the zone; (M) Anxiety index. (N to P) Effects of GnIH knockout on anxiety‐like behaviors during the light dark box following acute restraint stress: (N) Time spent in light box; (O) Time spent in dark box; (P) Percentage in the light box. (Q) Serum concentrations of CORT, NE and EPI in GnIH knockout mice following acute restraint stress. (R) TH , and VMAT2 mRNA levels in the locus coeruleus of GnIH knockout mice following acute restraint stress. (S to V) Adrenal mRNA ( TH , PNMT , NET , VMAT2 , SNAP25 , and synaptophysin ) and protein (TH, and Synaptophysin) levels of GnIH knockout mice following acute restraint stress. GAPDH and β‐tubulin was used as an internal control. Data are mean ± SEM (n = 10 biological replicates per group). Comparisons between two groups were performed using two‐sided unpaired Student's t ‐test with α = 0.05. For multiple comparisons, one‐way ANOVA followed by Tukey's post‐hoc test was used. Effect sizes (Cohen's d) were calculated for all significant comparisons. * p < 0.05, ** p < 0.01, GnIH Hetero or GnIH Homo vs. WT. # p < 0.05, ## p < 0.01, GnIH Hetero or GnIH Homo under RS vs. WT under RS. △ p < 0.05, △△ p < 0.01, WT vs. WT under RS. † p < 0.05, †† p < 0.01, GnIH Homo vs. GnIH Homo under RS.

Article Snippet: The following primary antibodies were used: β‐tubulin (Cat. 66240‐1‐Ig, proteintech, 1:2000 dilution; immunogen: recombinant human Tubulin‐beta protein; species reactivity: mouse), TH (Cat. sc‐25269, Santa Cruz, 1:100 dilution; immunogen: human TH amino acids 1–196; species reactivity: mouse), norepinephrine transporter (NET) (Cat. 680447, Zenbio, 1:1000 dilution; immunogen: a synthetic peptide of human NET; species reactivity: mouse), synaptosomal‐associated protein, 25 kDa (SNAP25) (Cat. R27294 , Zenbio, 1:1000 dilution; immunogen: human SNAP25; species reactivity: mouse), Syntaxin 1A (Cat. R389318, Zenbio, 1:1000 dilution; immunogen: human STX1A; species reactivity: mouse) and Synaptophysin (Cat. R380890, Zenbio, 1:1000 dilution; immunogen: human Synaptophysin amino acids 10–310; species reactivity: mouse).

Techniques: Knock-Out, Activation Assay, Control

Journal: Advanced Science

Article Title: Gonadotropin‐Inhibitory Hormone (GnIH) and Its Receptor GPR147 Mediates Acute Stress‐Induced Activation of the Locus Coeruleus‐SAM Axis

doi: 10.1002/advs.202518996

Figure Lengend Snippet: Effects of GPR147 Knockout on Anxiety‐like Behaviors and LC‐SAM Axis Activation in Mice under Acute Restraint Stress. (A to D) Effects of GPR147 knockout on anxiety‐like behaviors during the open field test following acute restraint stress: (A) Motion track and heat map; (B) Number of rearing; (C) Total locomotion; (D) Anxiety index. (E to I) Effects of GPR147 knockout on anxiety‐like behaviors during the elevated plus‐maze test following acute restraint stress: (E) Motion track and heat map; (F) Number of rearing; (G) Total locomotion; (H) Number of entries to the zone; (I) Anxiety index. (J to L) Effects of GPR147 knockout on anxiety‐like behaviors during the light dark box following acute restraint stress: (J) Time spent in light box; (K) Time spent in dark box; (L) Percentage in the light box. (M) Serum concentrations of CORT, NE and EPI in GPR147 knockout mice following acute restraint stress. (N and O) TH mRNA and TH protein levels in the locus coeruleus of GPR147 knockout mice following acute restraint stress. (P to R) Adrenal mRNA ( TH , PNMT , Syntaxin 1A and synaptophysin ) and protein (TH, and SNAP25) levels of GPR147 knockout mice following acute restraint stress. GAPDH and β‐tubulin was used as an internal control. Data are mean ± SEM (n = 10 biological replicates per group). Comparisons between two groups were performed using two‐sided unpaired Student's t ‐test with α = 0.05. For multiple comparisons, one‐way ANOVA followed by Tukey's post‐hoc test was used. Effect sizes (Cohen's d) were calculated for all significant comparisons. * p < 0.05, ** p < 0.01, GPR147 Hetero or GPR147 Homo vs. WT. # p < 0.05, ## p < 0.01, GPR147 Hetero or GPR147 Homo under RS vs. WT under RS. △ p < 0.05, △△ p < 0.01, WT vs. WT under RS. † p < 0.05, †† p < 0.01, GPR147 Homo vs. GPR147 Homo under RS.

Article Snippet: The following primary antibodies were used: β‐tubulin (Cat. 66240‐1‐Ig, proteintech, 1:2000 dilution; immunogen: recombinant human Tubulin‐beta protein; species reactivity: mouse), TH (Cat. sc‐25269, Santa Cruz, 1:100 dilution; immunogen: human TH amino acids 1–196; species reactivity: mouse), norepinephrine transporter (NET) (Cat. 680447, Zenbio, 1:1000 dilution; immunogen: a synthetic peptide of human NET; species reactivity: mouse), synaptosomal‐associated protein, 25 kDa (SNAP25) (Cat. R27294 , Zenbio, 1:1000 dilution; immunogen: human SNAP25; species reactivity: mouse), Syntaxin 1A (Cat. R389318, Zenbio, 1:1000 dilution; immunogen: human STX1A; species reactivity: mouse) and Synaptophysin (Cat. R380890, Zenbio, 1:1000 dilution; immunogen: human Synaptophysin amino acids 10–310; species reactivity: mouse).

Techniques: Knock-Out, Activation Assay, Control

Journal: Advanced Science

Article Title: Gonadotropin‐Inhibitory Hormone (GnIH) and Its Receptor GPR147 Mediates Acute Stress‐Induced Activation of the Locus Coeruleus‐SAM Axis

doi: 10.1002/advs.202518996

Figure Lengend Snippet: Effects of Acute Intracerebroventricular GnIH Injection on LC‐SAM Axis Activation and Norepinephrine Secretion in Mice. (A) Concentrations of CORT, NE, and EPI in serum following acute intracerebroventricular administration of GnIH. (B) Representative immunohistochemistry images and quantification of TH and ChAT immunoreactivity in the locus coeruleus after acute intracerebroventricular GnIH administration. Scale bar, 200/500 µm. (C and D): TH , NET , and VMAT2 mRNA and TH protein levels in the locus coeruleus after acute intracerebroventricular GnIH administration. (E) Representative immunohistochemistry images and quantification of TH and PNMT immunoreactivity in the adrenal gland after acute intracerebroventricular GnIH administration. Scale bar, 100/200 µm. (F to I): Adrenal mRNA ( TH , PNMT , NET , VMAT2 , SNAP25 , Syntaxin 1A, synaptophysin , and C‐FOS ) and protein (TH, SNAP25, Syntaxin 1A, Synaptophysin) levels after acute intracerebroventricular GnIH administration. (J) Concentrations of CORT, NE, and EPI in serum following acute intra‐LC injection of GnIH. (K) TH , NET , and VMAT2 mRNA levels in the LC after acute intra‐LC injection of GnIH. (L) Adrenal mRNA ( TH , PNMT , NET , VMAT2 , SNAP25 , Syntaxin 1A , and synaptophysin ) levels after acute intra‐LC injection of GnIH. LC: locus coeruleus; 4V: fourth ventricle; LH: spinal cord lateral horn. GAPDH and β‐tubulin was used as an internal control. Data are mean ± SEM (n = 10 biological replicates per group). Comparisons between two groups were performed using two‐sided unpaired Student's t ‐test with α = 0.05. For multiple comparisons, one‐way ANOVA followed by Tukey's post‐hoc test was used. Effect sizes (Cohen's d) were calculated for all significant comparisons. * p < 0.05, ** p < 0.01, compared to the control group.

Article Snippet: The following primary antibodies were used: β‐tubulin (Cat. 66240‐1‐Ig, proteintech, 1:2000 dilution; immunogen: recombinant human Tubulin‐beta protein; species reactivity: mouse), TH (Cat. sc‐25269, Santa Cruz, 1:100 dilution; immunogen: human TH amino acids 1–196; species reactivity: mouse), norepinephrine transporter (NET) (Cat. 680447, Zenbio, 1:1000 dilution; immunogen: a synthetic peptide of human NET; species reactivity: mouse), synaptosomal‐associated protein, 25 kDa (SNAP25) (Cat. R27294 , Zenbio, 1:1000 dilution; immunogen: human SNAP25; species reactivity: mouse), Syntaxin 1A (Cat. R389318, Zenbio, 1:1000 dilution; immunogen: human STX1A; species reactivity: mouse) and Synaptophysin (Cat. R380890, Zenbio, 1:1000 dilution; immunogen: human Synaptophysin amino acids 10–310; species reactivity: mouse).

Techniques: Injection, Activation Assay, Immunohistochemistry, Control

Journal: Advanced Science

Article Title: Gonadotropin‐Inhibitory Hormone (GnIH) and Its Receptor GPR147 Mediates Acute Stress‐Induced Activation of the Locus Coeruleus‐SAM Axis

doi: 10.1002/advs.202518996

Figure Lengend Snippet: Effects of Acute Intracerebroventricular GnIH Injection on LC‐SAM Axis Activation and Norepinephrine Secretion in Mice with Sympathetic Nerve Ablation (A) Serum levels of CORT, NE, and EPI following acute intracerebroventricular GnIH administration in sympathectomized mice. (B and C) TH , NET , and VMAT2 mRNA and TH protein levels in the locus coeruleus of sympathectomized mice after acute intracerebroventricular GnIH administration. (D to G) Expression of catecholamine biosynthesis enzyme mRNAs ( TH , PNMT , Dbh and Ddc ), catecholamine transporter mRNAs ( NET and VMAT2 ), synaptic vesicle protein mRNAs ( SNAP25 , Syntaxin 1A , Synaptophysin ) and protein (TH, SNAP25, Syntaxin 1A) levels in adrenal gland of sympathectomized mice after acute intracerebroventricular GnIH administration. GAPDH and β‐tubulin was used as an internal control. Data are mean ± SEM (n = 10 biological replicates per group). Comparisons between two groups were performed using two‐sided unpaired Student's t ‐test with α = 0.05. For multiple comparisons, one‐way ANOVA followed by Tukey's post‐hoc test was used. Effect sizes (Cohen's d) were calculated for all significant comparisons. * p < 0.05, ** p < 0.01, between the indicated groups.

Article Snippet: The following primary antibodies were used: β‐tubulin (Cat. 66240‐1‐Ig, proteintech, 1:2000 dilution; immunogen: recombinant human Tubulin‐beta protein; species reactivity: mouse), TH (Cat. sc‐25269, Santa Cruz, 1:100 dilution; immunogen: human TH amino acids 1–196; species reactivity: mouse), norepinephrine transporter (NET) (Cat. 680447, Zenbio, 1:1000 dilution; immunogen: a synthetic peptide of human NET; species reactivity: mouse), synaptosomal‐associated protein, 25 kDa (SNAP25) (Cat. R27294 , Zenbio, 1:1000 dilution; immunogen: human SNAP25; species reactivity: mouse), Syntaxin 1A (Cat. R389318, Zenbio, 1:1000 dilution; immunogen: human STX1A; species reactivity: mouse) and Synaptophysin (Cat. R380890, Zenbio, 1:1000 dilution; immunogen: human Synaptophysin amino acids 10–310; species reactivity: mouse).

Techniques: Injection, Activation Assay, Expressing, Control