Abstract 335: Vascular Histone Deacetylase Mediates Early Life Stress-Induced Endothelial Dysfunction

Chromatin remodeling is an important factor in the etiology of vascular pathologies. Also, early life stress (ELS) is linked to increased risk of vascular disease in adults. We used maternal separation with early weaning (MSEW) to study mechanisms of ELS-mediated adult vascular dysfunction in male C57BL/6J mice. Litters were subjected to maternal separation 4h/day (postnatal day (PD) 2-5) and 8h/day (PD6-16), and weaned at PD17. Control (CON) litters were undisturbed until weaning at PD21. Subsequent experiments were performed at 12 weeks old. MSEW blunted aortic ACh-mediated vasorelaxation (MSEW: 68% vs CON: 90%, p=0.01), while SNP-induced vasorelaxation was similar in CON and MSEW aortae. Apocynin (300 μM) and superoxide dismutase (100 U/mL) normalized MSEW-induced endothelial dysfunction. We hypothesize that ELS induces aortic endothelial dysfunction by increasing NADPH oxidase expression and/or decreasing nitric oxide synthase 3 (NOS3) expression. Aortic protein expression of NADPH oxidase subunit p67 was elevated in MSEW mice (45% increase from CON, n=11, p=0.02). NOS3 protein expression and NOS3 serine 1177 phosphorylation was not different between groups, indicating that NOS3 activation by phosphorylation does not contribute to ELS-induced endothelial dysfunction. We further hypothesize that chromatin modification mediates ELS-induced endothelial dysfunction. Aortic mRNA expressions of 84 chromatin modification enzymes (methyltransferases, demethylases, acetyltransferases, deacetylases) were assessed by qRT-PCR. Only histone deacetylase (HDAC) 1, 6 and 9 mRNA levels were significantly upregulated in MSEW aortae compared to CON (17%, 29% and 67% increase, respectively, p<0.05). However, only HDAC 9 protein expression was elevated in MSEW aortae (2 fold increase from CON, n=6, p=0.01). Accordingly, histone 3 lysine acetylation was slightly decreased in MSEW aortae (16% decrease from CON, n=6, p = 0.06). Pretreatment of aortae with an HDAC inhibitor, trichostatin A (TSA), normalized ACh-induced vasorelaxation in MSEW mice (MSEW: 68% vs MSEW + TSA: 88%, p=0.02), while not affecting ACh-induced vasorelaxation in CON mice. We conclude that ELS induces endothelial dysfunction, most likely, through an HDAC 9-mediated pathway.

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Abstract 57: Histone Deacetylase Inhibition Attenuates Early Life Stress-Induced Endothelial Dysfunction

Hypertension ◽

10.1161/hyp.60.suppl_1.a57 ◽

2012 ◽

Vol 60 (suppl_1) ◽

Author(s):

Dao H Ho ◽

Jennifer S Pollock

Keyword(s):

Blood Pressure ◽

Endothelial Dysfunction ◽

Histone Deacetylase ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Trichostatin A ◽

Early Life Stress ◽

Intercellular Adhesion Molecule ◽

Pressure Independent

Epidemiological studies show that early life stress (ELS) is linked to cardiovascular disease in adulthood. We used a model of maternal separation with early weaning (MSEW) to study the mechanisms of ELS-mediated adult vascular dysfunction in male C57BL/6J mice. MSEW litters were subjected to maternal separation 4h/day (postnatal day (PD) 2 to 5) and 8h/day (PD6 to 16), and weaned at PD17. Control (CON) litters were undisturbed until weaning at PD21. All subsequent experiments were performed in adult mice (12 weeks old). We hypothesized that MSEW increases vascular inflammation and endothelial dysfunction in male mice. Systolic blood pressure (tail-cuff) of MSEW mice was not different from CON mice (109.3 + 10.9 vs 116.7 + 20.8 mmHg, respectively). Circulating soluble intercellular adhesion molecule (CON: 333.5 + 19.4 vs MSEW: 406.2 + 23.1 ng/ml; p = 0.03) and macrophage colony stimulating factor (CON: 737.4 + 19.6 vs MSEW: 945.3 + 65.4 pg/ml; p = 0.01) were elevated by MSEW. Also, aortic adventitial macrophage infiltration was increased in mice exposed to MSEW (F4/80 immunostaining; CON: 2.8 + 2.3 vs MSEW: 7.0 + 2.2 cells/mm 2 ; p = 0.05). We performed wire myography on thoracic aortae to determine vasorelaxation with cumulative concentration-response curve to acetylcholine (ACh; 1 X 10 -9 M to 3 X 10 -5 M) and sodium nitroprusside (SNP; 1 X 10 -10 M to 3 X 10 -5 M). MSEW induced blunted ACh-mediated vasorelaxation (MSEW: 67.6 + 5.8 vs CON: 89.9 + 2.7 % of phenylephrine constriction (% of PE), p = 0.01), while SNP-induced vasorelaxation was similar in CON and MSEW mice. We further hypothesized that MSEW-induced endothelial dysfunction is mediated via increased histone deacetylase (HDAC) expression. Real-time quantitative PCR revealed upregulation of HDAC 1, 6 and 9 in aortae of MSEW mice (1.28 + 0.12, 1.28 + 0.18 and 1.65 + 0.05 fold change from CON, respectively, p < 0.05). Pretreatment with trichostatin A (TSA), an HDAC inhibitor, normalized ACh-induced vasorelaxation in aortae of MSEW mice (MSEW: 67.6 + 5.8 vs MSEW + TSA: 88.44 + 3.2 % of PE, p = 0.02), while not affecting ACh-induced vasorelaxation in aortae from CON mice (CON: 89.9 + 2.7 vs CON + TSA: 90.3 + 4.5 % of PE). We conclude that ELS induces blood pressure-independent endothelial dysfunction through an HDAC-mediated pathway.

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Exacerbated obesogenic response in female mice exposed to early life stress is linked to fat depot-specific upregulation of leptin protein expression

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00243.2020 ◽

2020 ◽

Vol 319 (5) ◽

pp. E852-E862

Author(s):

Jacqueline R. Leachman ◽

Mathew D. Rea ◽

Dianne M. Cohn ◽

Xiu Xu ◽

Yvonne N. Fondufe-Mittendorf ◽

...

Keyword(s):

Protein Expression ◽

Fat Mass ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Disease Risk ◽

Early Life Stress ◽

Female Mice ◽

Fat Depot ◽

Leptin Expression

Early life stress (ELS) is an independent risk factor for increased BMI and cardiometabolic disease risk later in life. We have previously shown that a mouse model of ELS, maternal separation and early weaning (MSEW), exacerbates high-fat diet (HF)-induced obesity only in adult female mice. Therefore, the aim of this study was to investigate 1) whether the short- and long-term effects of HF on leptin expression are influenced by MSEW in a sex-specific manner and 2) the potential epigenetic mechanisms underlying the MSEW-induced changes in leptin expression. After 1 wk of HF, both MSEW male and female mice displayed increased fat mass compared with controls ( P < 0.05). However, only MSEW female mice showed elevated leptin mRNA expression in gonadal white adipose tissue (gWAT; P < 0.05). After 12 wk of HF, fat mass remained increased only in female mice ( P < 0.05). Moreover, plasma leptin and both leptin mRNA and protein expression in gWAT were augmented in MSEW female mice compered to controls ( P < 0.05), but not in MSEW male mice. This association was not present in subcutaneous WAT. Furthermore, among 16 CpG sites in the leptin promoter, we identified three hypomethylated sites in tissue from HF-fed MSEW female mice compared with controls (3, 15, and 16, P < 0.05). These hypomethylated sites showed greater binding of key adipogenic factors such as PPARγ ( P < 0.05). Taken together, our study reveals that MSEW superimposed to HF increases leptin protein expression in a sex- and fat depot-specific fashion. Our data suggest that the mechanism by which MSEW increases leptin expression could be epigenetic.

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Abstract P197: Increased Serotonin In Visceral Adipose Tissue May Contribute To Stimulate Sensory Neurons Mediating Obesity Hypertension In Mice Exposed To Early Life Stress

Hypertension ◽

10.1161/hyp.78.suppl_1.p197 ◽

2021 ◽

Vol 78 (Suppl_1) ◽

Author(s):

Carolina Dalmasso ◽

Jacqueline Leachman ◽

Sundus Ghuneim ◽

Nermin Ahmed ◽

Jorge F Giani ◽

...

Keyword(s):

Adipose Tissue ◽

Protein Expression ◽

Sensory Neurons ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Tryptophan Hydroxylase ◽

Early Life Stress ◽

Trpv1 Channels ◽

Stimulation Of

Male C57BL/6J mice exposed to maternal separation and early weaning (MSEW), a mouse model of early life stress, display increased blood pressure (BP) and sympathetic activation compared to obese controls when fed a high fat diet (HF). Moreover, HF-fed MSEW males display exacerbated BP responses to the acute stimulation of the adipose afferent reflex (AAR) in epididymal white adipose tissue (eWAT). The aim of this study was to investigate the contribution of endogenous factors that could stimulate fat sensory neurons. MSEW and control (C) mice (n=8/group) were placed on a LF or HF (10% and 60% Kcal from fat, respectively) for 16 weeks. Then, serum obtained by decapitation and adipose tissue samples were collected to measure mRNA and protein expression of 15 factors and receptors known to activate sensory neurons. No differences were found across measurements on LF. Plasma AGT and AngII were decreased in HF-fed MSEW compared to C (AGT: 760±48 vs. 1267±161 ng/ml, p<0.05; AngII; 413±57 vs. 1082±340 pmol/l, p<0.07, Attoquant) and no differences were found in leptin (103±6 vs. 104±4 ng/ml, p<0.87). In eWAT, MSEW and C showed similar AGT (2.1±0.4 vs. 1.9±0.3 ng/ml per g tissue), AngII (1.7±0.2 vs. 2.3±0.5 pg AngII/mg tissue), ACE 1 activity (21.5±1.2 vs. 20.0±0.9 RFU/min/μg protein, p<0.33) and leptin (102.8±6.1 vs. 104.5±6.8 ng/mg of tissue, p<0.87). However, HF-fed MSEW showed increased eWAT mRNA expression of tryptophan hydroxylase 1 (Tph1), the rate limiting enzyme in serotonin (5-HT) synthesis (10.2±2.9 vs. 1.6±0.3 2 -ΔΔct , p<0.03). SERT-Tph1-MAO signaling pathway protein expression was activated, and fat serotonin concentration was also increased in eWAT from obese MSEW mice compared to C (16.58±1.5 vs. 8.5±2.1 ug/mg of tissue, p<0.01). Acute stimulation of eWAT with serotonin (10-6 M, 4 sites, 2 ul/site) tend to increase pressor response in MSEW mice (p<0.066, n=2-3). Unlike in female MSEW mice, our study demonstrates that MSEW does not increase circulating and tissue AGT, Ang II and leptin in male mice. Taken together, these data suggest that increased local serotonin could be endogenously sensitizing the sensory neurons in obese MSEW mice contributing to chronic AAR stimulation, directly via TRPV1 channels, or indirectly, via acid-sensing ion channels.

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Early life stress in male mice induces superoxide production and endothelial dysfunction in adulthood

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00016.2016 ◽

2016 ◽

Vol 310 (9) ◽

pp. H1267-H1274 ◽

Cited By ~ 10

Author(s):

Dao H. Ho ◽

Mariah L. Burch ◽

Benjamin Musall ◽

Jacqueline B. Musall ◽

Kelly A. Hyndman ◽

...

Keyword(s):

Nitric Oxide ◽

Cardiovascular Disease ◽

Endothelial Dysfunction ◽

Nadph Oxidase ◽

Life Stress ◽

Early Life ◽

Early Life Stress ◽

Superoxide Production ◽

Male Mice ◽

Oxide Synthase

Early life stress (ELS) is a risk for cardiovascular disease in adulthood although very little mechanistic insight is available. Because oxidative stress and endothelial dysfunction are major contributors to cardiovascular risk, we hypothesized that ELS induces endothelial dysfunction in adult male mice via increased superoxide production. Studies employed a mouse model of ELS, maternal separation with early weaning (MSEW), in which litters were separated from the dam for 4 h/day [ postnatal days (PD) 2–5] and 8 h/day (PD6-16), and weaned at PD17. Control litters remained undisturbed until weaning at PD21. When compared with control mice, thoracic aortic rings from adult male MSEW mice displayed significant endothelial dysfunction that was reversed by the superoxide scavenger, polyethylene glycol-superoxide dismutase (PEG-SOD). PEG-SOD-inhibitable superoxide production by aortae from MSEW mice was significantly greater than observed in control aortae, although unaffected by nitric oxide synthase inhibition, suggesting that uncoupled nitric oxide synthase was not responsible for the accelerated superoxide production. Aortic SOD expression, plasma SOD activity, and total antioxidant activity were similar in MSEW and control mice, indicating unaltered antioxidant capacity in MSEW mice. Increased expression of the NADPH oxidase subunits, NOX2 and NOX4, was evident in the aortae of MSEW mice. Moreover, endothelial dysfunction and superoxide production in MSEW mice was reversed with the NADPH oxidase inhibitor, apocynin, indicating increased NADPH oxidase-dependent superoxide production and endothelial dysfunction. The finding that MSEW induces superoxide production and endothelial dysfunction in adult mice may provide a mechanistic link between ELS and adult cardiovascular disease risk.

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Abstract 360: Early Life Stress Increases Hematocrit and Endothelin-1 in Mice

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.34.suppl_1.360 ◽

2014 ◽

Vol 34 (suppl_1) ◽

Author(s):

Dao H Ho ◽

Jennifer S Pollock

Keyword(s):

Endothelial Dysfunction ◽

Red Blood Cells ◽

Blood Cells ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Radioligand Binding ◽

Early Life Stress ◽

Male Mice ◽

Endothelin 1

In humans, early life stress (ELS) is an independent risk factor for adult cardiovascular disease (CVD). We have shown that mice subjected to ELS by maternal separation with early weaning (MSEW), develop vascular endothelial dysfunction in adulthood. A marker of endothelial dysfunction and CVD is high hematocrit, an abnormally elevated level of circulating red blood cells. Hematocrit is largely regulated by erythropoietin (EPO), a protein that is released predominantly from the kidney under conditions of hypoxia. We hypothesized that MSEW increases circulating EPO and hematocrit in adult male mice. We used the MSEW model in C57BL6J mice to study the mechanisms of ELS-mediated alteration in hematocrit. MSEW litters were subjected to maternal separation 4h/day (postnatal day (PD) 2-5) and 8h/day (PD6-16), and weaned at PD17. Control (CON) litters were undisturbed until weaning at PD21. At 13 weeks of age, blood was collected from CON and MSEW male mice by cardiac puncture and lung tissue was excised. Hematocrit of MSEW mice was significantly higher than CON mice (46.2 ± 0.03 vs 43.3 ± 0.03%, p = 0.004). Plasma EPO, as measured by ELISA, was elevated in MSEW mice, however not significantly (112.89 ± 51.32 vs 61.62 ± 20.73 pg/ml, p = 0.06). We further hypothesized that MSEW enhances circulating endothelin-1 (ET-1) levels, a vasoactive peptide regulated by hypoxia and EPO. We found that plasma ET-1 was significantly increased in MSEW mice compared to CON (1.55 ± 0.41 vs 1.26 ± 0.23 pg/ml, p = 0.02). Endothelin receptor type A and B density and binding in lung, as measured by radioligand binding, was not different between groups, suggesting that increased circulating ET-1 in MSEW mice was not due to decreased ET-1 clearance in the lungs and most likely is due to increased production of ET-1. Taken together, our data suggest that MSEW-induced endothelial dysfunction may be mediated by an interplay of increased circulating red blood cells and elevated ET-1 production. Further studies are necessary to determine the exact role of these factors in this phenomenon.

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Early life stress induces endothelial dysfunction in a mouse model of maternal separation

The FASEB Journal ◽

10.1096/fasebj.26.1_supplement.1101.2 ◽

2012 ◽

Vol 26 (S1) ◽

Author(s):

Dao H. Ho ◽

Megan L. Yu ◽

Catalina Bazacliu ◽

Jennifer S. Pollock

Keyword(s):

Endothelial Dysfunction ◽

Mouse Model ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Early Life Stress

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Cocaine-Induced Impulsivity is Differentially Expressed in Male and Female Mice Exposed to Maternal Separation and is Associated with Alterations in AMPA Receptors Subunits

10.1101/2020.06.05.136812 ◽

2020 ◽

Author(s):

Adriana Castro-Zavala ◽

Ana Martin-Sanchez ◽

Larisa Montalvo-Martínez ◽

Alberto Camacho-Morales ◽

Olga Valverde

Keyword(s):

Protein Expression ◽

Ampa Receptors ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Early Life Stress ◽

Male Mice ◽

Male And Female ◽

Female Mice ◽

Cocaine Seeking

ABSTRACTImpulsivity is a key trait in the diagnosis of major depressive disorder (MDD) and substance use disorder (SUD). MDD is a chronic illness characterized by sadness, insomnia, and loss of interest. SUD is a chronic and relapsing disorder characterized by the consumption of drugs despite their negative consequences. Among drugs of abuse, cocaine is the most consumed psychostimulant. Animal studies demonstrated that increased impulsivity predicts predisposition to acquire cocaine self-administration (SA) behaviour with an increased cocaine-intake. Moreover, early-life stress represents a vulnerability factor to develop depressive disorders and drug addiction. Maternal separation with early weaning (MSEW) is an animal model that allows examining the impact of early-life stress on cocaine abuse. In this study, we aimed to explore changes in MSEW-induced impulsivity to determine potential associations between depression-like and cocaine-seeking behaviours in male and female mice. We also evaluated possible alterations in the AMPA receptors (AMPArs) composition and glutamatergic neurotransmission. We exposed mice to MSEW and the behavioural tests were performed during adulthood. Moreover, GluA1, GluA2 mRNA and protein expression were evaluated in the medial Prefrontal Cortex (mPFC). Results showed higher impulsive cocaine-seeking in females, independently the MSEW, as well as an increase in GluA1 and GluA2 protein expression. Moreover, MSEW induced downregulation of Gria2 and increased the GluA1/GluA2 ratio, only in male mice. In conclusion, female mice expressed higher mPFC glutamatergic function, which potentiated their impulsivity during cocaine SA. Also, data indicated that MSEW alters glutamatergic function in mPFC of male mice, increasing the glutamatergic excitability.

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Early-Life Stress Modulates Gut Microbiota and Peripheral and Central Inflammation in a Sex-Dependent Manner

International Journal of Molecular Sciences ◽

10.3390/ijms22041899 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1899 ◽

Cited By ~ 1

Author(s):

Hae Jeong Park ◽

Sang A. Kim ◽

Won Sub Kang ◽

Jong Woo Kim

Keyword(s):

Gut Microbiota ◽

Relative Abundance ◽

Life Stress ◽

Early Life ◽

Maternal Separation ◽

Early Life Stress ◽

Female Rats ◽

Rrna Gene ◽

Dependent Manner ◽

Male And Female Rats

Recent studies have reported that changes in gut microbiota composition could induce neuropsychiatric problems. In this study, we investigated alterations in gut microbiota induced by early-life stress (ELS) in rats subjected to maternal separation (MS; 6 h a day, postnatal days (PNDs) 1–21), along with changes in inflammatory cytokines and tryptophan-kynurenine (TRP-KYN) metabolism, and assessed the differences between sexes. High-throughput sequencing of the bacterial 16S rRNA gene showed that the relative abundance of the Bacteroides genus was increased and that of the Lachnospiraceae family was decreased in the feces of MS rats of both sexes (PND 56). By comparison, MS increased the relative abundance of the Streptococcus genus and decreased that of the Staphylococcus genus only in males, whereas the abundance of the Sporobacter genus was enhanced and that of the Mucispirillum genus was reduced by MS only in females. In addition, the levels of proinflammatory cytokines were increased in the colons (IFN-γ and IL-6) and sera (IL-1β) of the male MS rats, together with the elevation of the KYN/TRP ratio in the sera, but not in females. In the hippocampus, MS elevated the level of IL-1β and the KYN/TRP ratio in both male and female rats. These results indicate that MS induces peripheral and central inflammation and TRP-KYN metabolism in a sex-dependent manner, together with sex-specific changes in gut microbes.

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