Journal: bioRxiv

Article Title: Hippocampus oxytocin signaling promotes prosocial eating in rats

doi: 10.1101/2024.01.03.574101

Figure Lengend Snippet: (A) Diagram depicting viral vector-mediated OXTR shRNA for chronic knockdown of OXTR expression in the HPCd. (B-C) Following the infusion of scrambled sequence control (scrmb) or OXTR shRNA AAVs, dentate gyrus target sites and viral-induced GFP expression were confirmed using immunohistochemistry (representative photomicrographs from each group depicted). (D) Quantification of relative OXTR mRNA expression showed a significant reduction in knockdown animals of approximately 70-80% relative to controls (control n=7; KD n=7). (E-G) During the training phase of the social eating procedure, OXTR KD animals consumed significantly less food within the hour in the social arena compared to controls, with a trend towards a reduction in first meal size (p=0.057) but no effect on meal frequency. (H-J ) Control, but not OXTR KD animals consumed more food within the hour-long test in the presence of a familiar vs. an unfamiliar conspecific; an effect driven by an increased 1 st meal size with no change in meal frequency. (K-M) Knockdown of dorsal hippocampal oxytocin receptors did not yield long-term changes in daily caloric intake or body weight under isolated conditions in the home cage. (Between-subjects design for group; control n=6; OXTR KD n=9; Data are means ± SEM; *p<0.05; Abbreviations, DGmo = dentate gyrus molecular layer, DGsg = dentate gyrus granule layer, DGpo = dentate gyrus polymorph layer).

Article Snippet: OXTR mRNA levels were quantified using Taqman gene expression kits (OXTR: Rn00563503_m1; GapDH: Rn01775763_g1; Life Technologies) and PCR reagents (Applied Biosystems). qPCR was conducted using an Eppendorf Mastercycler ep realplex2 and the comparative threshold cycle method was used to quantify relative mRNA expression.

Techniques: Plasmid Preparation, shRNA, Knockdown, Expressing, Sequencing, Control, Immunohistochemistry, Isolation

Journal: bioRxiv

Article Title: Hippocampus oxytocin signaling promotes prosocial eating in rats

doi: 10.1101/2024.01.03.574101

Figure Lengend Snippet: (A-C) During the training phase of the social eating procedure, OXTR KD animals saw a reduction in average meal size during the hour in the social arena compared to controls while there was no significant difference in either 1 st meal duration or average meal duration. (D-I ) Control, but not OXTR KD animals had a larger average meal size during the hour-long test in the presence of a familiar vs. an unfamiliar conspecific, while showing no effect of 1 st meal duration, average meal duration, bout frequency, average bout size or average bout duration. (J-P) Knockdown of dorsal hippocampal oxytocin receptors did not yield long-term changes in daily meal parameters or body weight under isolated conditions in the home cage. (Between subjects control n=6; OXTR KD n=9; Data are means ± SEM; *p<0.05).

Article Snippet: OXTR mRNA levels were quantified using Taqman gene expression kits (OXTR: Rn00563503_m1; GapDH: Rn01775763_g1; Life Technologies) and PCR reagents (Applied Biosystems). qPCR was conducted using an Eppendorf Mastercycler ep realplex2 and the comparative threshold cycle method was used to quantify relative mRNA expression.

Techniques: Control, Knockdown, Isolation

Journal: bioRxiv

Article Title: Hippocampus oxytocin signaling promotes prosocial eating in rats

doi: 10.1101/2024.01.03.574101

Figure Lengend Snippet: (A) Diagram depicting STFP procedure in which an experimental or observer rat is exposed to a novel food flavor from the breath of a demonstrator that has recently consumed the flavored chow in a separate room. 24 hours after exposure to demonstrator rats, observer rats are tested in the two-choice preference consumption test. (B) There was no difference between Control and OXTR KD rats in the time spent investigating the demonstrator during social interaction. (C) Control animals successfully demonstrated a significant preference for the flavor their demonstrator had consumed, whereas OXTR KD rats showed no flavor preference. (D) The total amount of food consumed during the preference test did not differ by group. (Between-subjects design for group; control n=6; OXTR KD n=9; Data are means ± SEM; *p<0.05, ****p<0.0001).

Article Snippet: OXTR mRNA levels were quantified using Taqman gene expression kits (OXTR: Rn00563503_m1; GapDH: Rn01775763_g1; Life Technologies) and PCR reagents (Applied Biosystems). qPCR was conducted using an Eppendorf Mastercycler ep realplex2 and the comparative threshold cycle method was used to quantify relative mRNA expression.

Techniques: Control

Journal: bioRxiv

Article Title: Hippocampus oxytocin signaling promotes prosocial eating in rats

doi: 10.1101/2024.01.03.574101

Figure Lengend Snippet: (A) Sociability is assessed by placing experimental animals into an arena for 5 min with an empty enclosure and another containing a stimulus animal. The time spent investigating each enclosure is measured. (B-C) Both control and HPCd OXTR KD animals spent significantly more time investigating the stimulus animal over the empty enclosure, indicating normal sociability in both groups. (D) Following an interval of 30 min, animals are placed back into the arena to assess social recognition memory, now with the previously experienced “familiar” stimulus animal or a new “novel” animal. (E-F) Control animals spent more time investigating the novel stimulus animal while the HPCd OXTR KD animals did not. (G) Animals were also tested in novel object recognition task to assess non-social recognition memory. (H-I) Control and OXTR KD groups performed similarly in the novel object recognition test. (Between-subjects for group; control n=7; OXTR KD n=7; Data are means ± SEM; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

Article Snippet: OXTR mRNA levels were quantified using Taqman gene expression kits (OXTR: Rn00563503_m1; GapDH: Rn01775763_g1; Life Technologies) and PCR reagents (Applied Biosystems). qPCR was conducted using an Eppendorf Mastercycler ep realplex2 and the comparative threshold cycle method was used to quantify relative mRNA expression.

Techniques: Control

Journal: Immunobiology

Article Title: Clostridium butyricum attenuates LPS-induced myocardial injury in septic mice by modulating CD4 + CD25 + FOXP3 + Treg.

doi: 10.1016/j.imbio.2024.152857

Figure Lengend Snippet: Fig. 2. Inflammation in septic myocardial injury mice with marked changes in markers of myocardial injury. (A-C) After the successful construction of a mouse model of sepsis induced myocardial injury, plasma was diluted three-fold and ELISA was used to detect changes in myocardial injury markers and inflammatory markers in the LPS group and control group, respectively (n = 5 per group). The main biomarkers for myocardial injury are cTnI and LDH, while inflammatory biomarkers are IL-1β, IL-6, and TNF-α. A: Changes in cTnI levels between two groups. B: Changes in LDH levels between two groups. C: Changes in IL-1β, IL-6, and TNF-α levels between two groups. D: qPCR assay for mouse heart tissue mRNA levels of inflammatory factors IL-1β, IL-6, and TNF-α between two groups. n represents number of animals, *P represents P < 0.05; **P represents P < 0.01; ***P represents P < 0.001; ****P represents P < 0.0001;

Article Snippet: The standard preparation and related experimental steps were performed according to the instruction manual of the ELISA kits for IL-6 (Sigma-Aldrich #RAB0308), IL-1β (Sigma-Aldrich #RAB0274), TNF-α (Sigma-Aldrich #RAB0477), cTnI (MM #0791 M1), and LDH (Cusabio #CSB-E11723m).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control

Journal: Immunobiology

Article Title: Clostridium butyricum attenuates LPS-induced myocardial injury in septic mice by modulating CD4 + CD25 + FOXP3 + Treg.

doi: 10.1016/j.imbio.2024.152857

Figure Lengend Snippet: Fig. 5. Clostridium butyricum significantly reduced serum inflammation and myocardial markers in mice with septic myocardial injury. (A-C) After continuous gavage of Clostridium butyricum for 28 days and successful construction of a mouse model of sepsis induced myocardial injury, plasma was diluted three-fold and ELISA was used to detect changes in myocardial injury markers and inflammatory markers in the LPS group and control group, respectively (n = 5 per group). The main biomarkers for myocardial injury are cTnI and LDH, while inflammatory biomarkers are IL-1β, IL-6, and TNF-α. A: Changes in cTnI levels between two groups. B: Changes in LDH levels between two groups. C: Changes in IL-1β, IL-6, and TNF-α levels between two groups. D: qPCR assay for mouse heart tissue mRNA levels of inflammatory factors IL-1β, IL-6, and TNF-α between two groups. n represents number of animals, *P represents P < 0.05; **P represents P < 0.01; ***P represents P < 0.001; ****P represents P < 0.0001;

Article Snippet: The standard preparation and related experimental steps were performed according to the instruction manual of the ELISA kits for IL-6 (Sigma-Aldrich #RAB0308), IL-1β (Sigma-Aldrich #RAB0274), TNF-α (Sigma-Aldrich #RAB0477), cTnI (MM #0791 M1), and LDH (Cusabio #CSB-E11723m).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control

Journal: Thyroid

Article Title: Decreased Expression of Hepatic Low-Density Lipoprotein Receptor–Related Protein 1 in Hypothyroidism: A Novel Mechanism of Atherogenic Dyslipidemia in Hypothyroidism

doi: 10.1089/thy.2012.0457

Figure Lengend Snippet: Change in hepatic LRP1 expression in a hypothyroidism animal model. C57BL/6 mice were fed a normal diet (control group, n=6) or a low-iodine diet supplemented with 0.15% propylthiouracil (PTU/LI group, n=4) for 4 weeks. (A, B) Western blot analysis of LRP1 (β-chain) in liver samples. (C) RT-PCR quantification of LRP1 mRNA in liver samples. Results are normalized with β-actin protein or mRNA levels and are expressed as ratios relative to the control group. Data are mean±SE. *p<0.05 vs. control group. RT-PCR, real-time polymerase chain reaction; LRP1, low-density lipoprotein receptor–related protein 1; SE, standard error.

Article Snippet: Quantitative real-time polymerase chain reaction Quantitative real-time polymerase chain reaction analysis was performed using a TaqMan assay kit for LRP1 (Hs00233856_m1, Mm00464608_m1) and an ABI 7500 instrument (Applied Biosystems, Foster City, CA). β-Actin (Hs99999903_m1, Mm00607939_s1) was used as an invariant control.

Techniques: Expressing, Animal Model, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: Thyroid

Article Title: Decreased Expression of Hepatic Low-Density Lipoprotein Receptor–Related Protein 1 in Hypothyroidism: A Novel Mechanism of Atherogenic Dyslipidemia in Hypothyroidism

doi: 10.1089/thy.2012.0457

Figure Lengend Snippet: Immunohistochemistry of LRP1 in the liver samples. L C57BL/6 mice were fed a normal diet (control group, n=6) or a low-iodine diet supplemented with 0.15% propylthiouracil (PTU/LI group, n=4) for 4 weeks. Three samples of each group are presented and LRP1 expression is shown by brown color. Bars indicate 100 μm. Color images are available online at www.liebertpub.com/thy

Article Snippet: Quantitative real-time polymerase chain reaction Quantitative real-time polymerase chain reaction analysis was performed using a TaqMan assay kit for LRP1 (Hs00233856_m1, Mm00464608_m1) and an ABI 7500 instrument (Applied Biosystems, Foster City, CA). β-Actin (Hs99999903_m1, Mm00607939_s1) was used as an invariant control.

Techniques: Immunohistochemistry, Control, Expressing

Journal: Thyroid

Article Title: Decreased Expression of Hepatic Low-Density Lipoprotein Receptor–Related Protein 1 in Hypothyroidism: A Novel Mechanism of Atherogenic Dyslipidemia in Hypothyroidism

doi: 10.1089/thy.2012.0457

Figure Lengend Snippet: Change in hepatic LRP1 expression after T3 treatment in animals. C57BL/6 mice were fed a low-iodine diet supplemented with 0.15% propylthiouracil (n=11). Various doses of T3 (0, 30, and 150 μg/kg of body weight) were administered daily to the mice through intraperitoneal injection for 7 days. (A, B) Western blot analysis of LRP1 (β-chain) in liver samples. (C) RT-PCR quantification of LRP1 mRNA in liver samples. Results are normalized with β-actin protein or mRNA levels and are expressed as ratios relative to 0 μg/kg. Data are mean±SE. *p<0.05 vs. 0 hour. T3, triiodothyronine.

Article Snippet: Quantitative real-time polymerase chain reaction Quantitative real-time polymerase chain reaction analysis was performed using a TaqMan assay kit for LRP1 (Hs00233856_m1, Mm00464608_m1) and an ABI 7500 instrument (Applied Biosystems, Foster City, CA). β-Actin (Hs99999903_m1, Mm00607939_s1) was used as an invariant control.

Techniques: Expressing, Injection, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Thyroid

Article Title: Decreased Expression of Hepatic Low-Density Lipoprotein Receptor–Related Protein 1 in Hypothyroidism: A Novel Mechanism of Atherogenic Dyslipidemia in Hypothyroidism

doi: 10.1089/thy.2012.0457

Figure Lengend Snippet: The effect of T3 on LRP1 expression in HepG2 cells. HepG2 cells incubated in CSS or unstripped FBS for 24 hours were treated with the indicated concentrations of T3 for 48 hours. (A) Western blot analysis of LRP1 (β-chain) in HepG2 cells (n=5 in each group). (B) Western blot analysis of LRP1 (β-chain) in HepG2 cells incubated in CSS (n=5 in each group). (C) RT-PCR quantification of LRP1 mRNA in HepG2 cells incubated in CSS (n=5 in each group). Results are normalized with β-actin protein or mRNA levels and are expressed as ratios relative to T3 0 nM in CSS. Data are mean±SE. *p<0.05 vs. T3 0 nM in CSS; †p<0.05 vs. T3 0.5 nM in CSS. CSS, charcoal-stripped fetal bovine serum; FBS, fetal bovine serum.

Article Snippet: Quantitative real-time polymerase chain reaction Quantitative real-time polymerase chain reaction analysis was performed using a TaqMan assay kit for LRP1 (Hs00233856_m1, Mm00464608_m1) and an ABI 7500 instrument (Applied Biosystems, Foster City, CA). β-Actin (Hs99999903_m1, Mm00607939_s1) was used as an invariant control.

Techniques: Expressing, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Thyroid

Article Title: Decreased Expression of Hepatic Low-Density Lipoprotein Receptor–Related Protein 1 in Hypothyroidism: A Novel Mechanism of Atherogenic Dyslipidemia in Hypothyroidism

doi: 10.1089/thy.2012.0457

Figure Lengend Snippet: Uptake of lipid-conjugated ApoE3 by T3 in HepG2 cells. HepG2 cells incubated in CSS for 24 hours were treated with the indicated concentrations of T3 for 48 hours. Human recombinant ApoE3 conjugated with lipid was added to the culture medium and cells were incubated for 1 hour. (A) Western blot analysis of ApoE3 in HepG2 cells incubated with or without added ApoE3. (B) Western blot analysis of ApoE3 in HepG2 cells transfected with nontargeting negative siRNA (siCTRL) and siRNA targeting LRP1 (siLRP1). (C) Western blot analysis of ApoE3 in HepG2 cells transfected with siCTRL and siRNA targeting the LDL receptor (siLDLR). The human recombinant receptor–associated protein was used as a functional blocker of LRP1 and LDLR. The human recombinant receptor–associated protein was added to the culture medium before adding ApoE3. Three independent experiments were performed for the representative figures. siRNA, small interfering RNA; LDL, low-density lipoprotein; LDLR, LDL receptor.

Article Snippet: Quantitative real-time polymerase chain reaction Quantitative real-time polymerase chain reaction analysis was performed using a TaqMan assay kit for LRP1 (Hs00233856_m1, Mm00464608_m1) and an ABI 7500 instrument (Applied Biosystems, Foster City, CA). β-Actin (Hs99999903_m1, Mm00607939_s1) was used as an invariant control.

Techniques: Incubation, Recombinant, Western Blot, Transfection, Functional Assay, Small Interfering RNA

Journal: PLoS ONE

Article Title: Regulation of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Protein Expression by MiR-34a in Sporadic Somatotropinomas

doi: 10.1371/journal.pone.0117107

Figure Lengend Snippet: TaqMan assays used in real-time qPCR quantification.

Article Snippet: RT-qPCR was performed with the TaqMan system using ready made AIP-probe primer kits (Hs_00610222_m1, Rn_00597273-m1, Life Technologies).

Techniques:

Journal: PLoS ONE

Article Title: Regulation of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Protein Expression by MiR-34a in Sporadic Somatotropinomas

doi: 10.1371/journal.pone.0117107

Figure Lengend Snippet: Demographic, radiological, biochemical and pathological characteristics of the patients with acromegaly.

Article Snippet: RT-qPCR was performed with the TaqMan system using ready made AIP-probe primer kits (Hs_00610222_m1, Rn_00597273-m1, Life Technologies).

Techniques: Biomarker Discovery, Control, Expressing

Journal: PLoS ONE

Article Title: Regulation of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Protein Expression by MiR-34a in Sporadic Somatotropinomas

doi: 10.1371/journal.pone.0117107

Figure Lengend Snippet: A and B—Examples of low AIP expression; C and D: Examples of high AIP expression; E—Normal human pituitary staining with omitting primary antibody (negative control); F—Normal human pituitary staining with AIP (positive control); Scale bar = 1000 μm.

Article Snippet: RT-qPCR was performed with the TaqMan system using ready made AIP-probe primer kits (Hs_00610222_m1, Rn_00597273-m1, Life Technologies).

Techniques: Expressing, Staining, Negative Control, Positive Control

Journal: PLoS ONE

Article Title: Regulation of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Protein Expression by MiR-34a in Sporadic Somatotropinomas

doi: 10.1371/journal.pone.0117107

Figure Lengend Snippet: The 11 miRNAs selected to analyse the expression level in patients with low or high AIP protein expression and the criteria for selection.

Article Snippet: RT-qPCR was performed with the TaqMan system using ready made AIP-probe primer kits (Hs_00610222_m1, Rn_00597273-m1, Life Technologies).

Techniques: Expressing, Selection

Journal: PLoS ONE

Article Title: Regulation of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Protein Expression by MiR-34a in Sporadic Somatotropinomas

doi: 10.1371/journal.pone.0117107

Figure Lengend Snippet: GH3 cells were transfected with plasmids containing pGL3- AIP -3’UTR WT and constructs with mutation in SITE A, SITE B, SITE C and SITE A+C and co-transfected with miR-34a or scrambled control. GH3 cells transfected with empty pGL3 vector and miR-34a or its scrambled control. Data are shown as Firefly/Renilla activity ratios compared to that of the scrambled control transfected cells. Mean ±SEM, *, P <0.05, ***, P <0.001.

Article Snippet: RT-qPCR was performed with the TaqMan system using ready made AIP-probe primer kits (Hs_00610222_m1, Rn_00597273-m1, Life Technologies).

Techniques: Transfection, Construct, Mutagenesis, Control, Plasmid Preparation, Activity Assay

Journal: PLoS ONE

Article Title: Regulation of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Protein Expression by MiR-34a in Sporadic Somatotropinomas

doi: 10.1371/journal.pone.0117107

Figure Lengend Snippet: Effect of miR-34a on endogenous AIP mRNA expression in HEK293 (D) and in GH3 (E) cells 48 hours after transfection measured by RT-qPCR. Data are shown as mean±SEM, **, P <0.01 ***, P <0.001.

Article Snippet: RT-qPCR was performed with the TaqMan system using ready made AIP-probe primer kits (Hs_00610222_m1, Rn_00597273-m1, Life Technologies).

Techniques: Expressing, Transfection, Quantitative RT-PCR