GABA release from central amygdala neurotensin neurons differentially modulates ethanol consumption in male and female mice.

The central nucleus of the amygdala is known to play key roles in alcohol use and affect. Neurotensin neurons in the central nucleus of the amygdala have been shown to regulate alcohol drinking in male mice. However, little is known about which neurotransmitters released by these cells drive alcohol consumption or whether these cells drive alcohol consumption in female mice. Here we show that knockdown of GABA release from central amygdala neurotensin neurons using a Nts-cre-dependent vGAT-shRNA-based AAV strategy reduces alcohol drinking in male, but not female, mice. This manipulation did not impact avoidance behavior, except in a fasted novelty-suppressed feeding test, in which vGAT shRNA mice demonstrated increased latency to feed on a familiar high-value food reward, an effect driven by male mice. In contrast, vGAT shRNA female mice showed heightened sensitivity to thermal stimulation. These data show a role for GABA release from central amygdala neurotensin neurons in modulating consumption of rewarding substances in different motivational states.

Voltage-gated potassium channels control extended access cocaine seeking: a role for nucleus accumbens astrocytes.

Dopaminergic signaling in the nucleus accumbens shell (NAc) regulates neuronal activity relevant to reward-related learning, including cocaine-associated behaviors. Although astrocytes respond to dopamine and cocaine with structural changes, the impact of dopamine and cocaine on astrocyte functional plasticity has not been widely studied. Specifically, behavioral implications of voltage-gated channel activity in the canonically non-excitable astrocytes are not known. We characterized potassium channel function in NAc astrocytes following exposure to exogenous dopamine or cocaine self-administration training under short (2 h/day) and extended (6 h/day) access schedules. Electrophysiological, Ca2+ imaging, mRNA, and mass spectrometry tools were used for molecular characterization. Behavioral effects were examined after NAc-targeted microinjections of channel antagonists and astroglial toxins. Exogenous dopamine increased activity of currents mediated by voltage-gated (Kv7) channels in NAc astrocytes. This was associated with a ~5-fold increase in expression of Kcnq2 transcript level in homogenized NAc micropunches. Matrix-assisted laser desorption/ionization mass spectrometry revealed increased NAc dopamine levels in extended access, relative to short access, rats. Kv7 inhibition selectively increased frequency and amplitude of astrocyte intracellular Ca2+ transients in NAc of extended access rats. Inhibition of Kv7 channels in the NAc attenuated cocaine-seeking in extended access rats only, an effect that was occluded by microinjection of the astrocyte metabolic poison, fluorocitrate. These results suggest that voltage-gated K+ channel signaling in NAc astrocytes is behaviorally relevant, support Kv7-mediated regulation of astrocyte Ca2+ signals, and propose novel mechanisms of neuroglial interactions relevant to drug use.

Does WeChat use intensity influence Chinese college students' mental health through social use of WeChat, entertainment use of WeChat, and bonding social capital?

Background: Previous research notes that the usage of WeChat is significantly related to individuals' mental health, but the underlying mechanism is still not completely discovered. The present study aimed to explore the sequential mediating roles of WeChat use motivations and bonding social capital on the effects of WeChat use intensity on mental health in Chinese college students. Method: The present study adopted an online survey with a total of 487 Chinese college students. Correlation analysis and serial mediation analysis were measured by process regarding the hypothesis. Results: The study presented findings indicating that WeChat use intensity had both direct and indirect impacts on the levels of life satisfaction and loneliness experienced by college students in China. Specifically, the utilization of WeChat for social motivation and entertainment motivation was found to have a suppressive effect on the relationship between the intensity of WeChat usage and individuals' life satisfaction. The association between the intensity of WeChat usage and mental health outcomes (life satisfaction and loneliness) was found to be mediated by bonding social capital. Furthermore, the association between the intensity of WeChat usage and mental health was found to be mediated by the sequential mediation effects of using WeChat for social motivation and bonding social capital, as well as the sequential mediation effects of using WeChat for entertainment motivation and bonding social capital. Conclusion: Our findings provide implications for policymakers and social workers regarding renovating the perceptions of the relationships between WeChat use intensity and overall mental health. Specifically, practical online activities and services of SNSs are recommended to be designed for meeting social and recreational gratifications and boosting bonding social capital, which in turn promotes psychological wellbeing.

GABA Release From Central Amygdala Neurotensin Neurons Differentially Modulates Reward and Consummatory Behavior in Male and Female Mice.

The central nucleus of the amygdala is known to play key roles in alcohol use and affect. Neurotensin neurons in the central nucleus of the amygdala have been shown to regulate alcohol drinking in male mice. However, little is known about which neurotransmitters released by these cells drive alcohol consumption or whether these cells drive alcohol consumption in female mice. Here we show that knockdown of GABA release from central amygdala neurotensin neurons using a Nts-cre-dependent vGAT-shRNA-based AAV strategy reduces alcohol drinking in male, but not female, mice. This manipulation did not impact avoidance behavior, except in a fasted novelty-suppressed feeding test, in which vGAT shRNA mice demonstrated increased latency to feed on a familiar high-value food reward, an effect driven by male mice. In contrast, vGAT shRNA female mice showed heightened sensitivity to thermal stimulation. These data show a role for GABA release from central amygdala neurotensin neurons in modulating consumption of rewarding substances in different motivational states.

Study of Calcium Signaling in Astrocytes with a Novel Endoplasmic Reticulum-Targeted GCaMP Sensor.

The endoplasmic reticulum (ER), the major organelle for the storage of Ca2+ , maintains a concentration of Ca2+ much higher than in the cytosol or other subcellular organelles, such as the mitochondria. A variety of tools have been developed for measuring Ca2+ activity in neuronal and glial cells, but most of these sensors target either the plasma membrane (PM) or the cytosol. Though these sensors are important for measuring Ca2+ transients, they lack the capability to measure activity in the periphery of the ER or to measure low-amplitude events resulting from Ca2+ exchange between the ER and other organelles, such as the mitochondria. We recently developed an ER-targeted GCaMP6f anchored to the cytosolic side of the ER that can measure minute calcium exchange occurring in this region. In this article, we discuss detailed methods to characterize the ER-GCaMP6f sensor, utilize it for calcium imaging in cultured astrocytes, and assess its expression and calcium imaging in astrocytes in rodent brains. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Expression and characterization of ER-GCaMP6f Support Protocol 1: ER-GCaMP6f-expressing stable cell line generation Basic Protocol 2: In vitro calcium imaging with ER-GCaMP6f Support Protocol 2: Imaging of drug-induced calcium activity Alternate Protocol 1: Transduction of astrocytes with ER-GCaMP6f AAV Alternate Protocol 2: Calcium imaging of astrocytes with Fluo-4 AM Basic Protocol 3: In vivo ER-GCaMP6f expression and slice calcium imaging Support Protocol 3: Pharmacological studies with 2-APB in brain slices.

ER-GCaMP6f: An Endoplasmic Reticulum-Targeted Genetic Probe to Measure Calcium Activity in Astrocytic Processes.

Ca2+ is a major second messenger involved in cellular and subcellular signaling in a wide range of cells, including astrocytes, which use calcium ions to communicate with other cells in the brain. Even though a variety of genetically encoded Ca2+ indicators have been developed to study astrocyte calcium signaling, understanding the dynamics of endoplasmic reticulum calcium signaling is greatly limited by the currently available tools. To address this, we developed an endoplasmic reticulum-targeted calcium indicator, ER-GCaMP6f, which is anchored to the cytosolic side of the organelle and measures signaling that occurs in close proximity to the endoplasmic reticulum of astrocytes. Using a combination of confocal and super-resolution microscopy techniques, we demonstrate the localization of the indicator in the endoplasmic reticulum in both cell soma and processes of astrocytes. Combining ER-GCaMP6f with total internal reflection fluorescence microscopy, we show that Ca2+ fluctuations in small astrocytic processes can be detected, which are otherwise not observable with existing indicators and standard wide-field and confocal techniques. We also compared the ER-GCaMP6f indicator against currently used plasma membrane-tethered and cytosolic GCaMP6f indicators. ER-GCaMP6f identifies dynamics in calcium signaling of endoplasmic reticulum resident receptors that are missed by plasma membrane-anchored indicators. We also generated an adeno-associated virus (AAV5) and demonstrate that ER-GCaMP6f can be expressed in vivo and by measured calcium activity in brain slices. ER-GCaMP6f provides a powerful tool to study calcium signaling in close proximity to the endoplasmic reticulum in astrocyte cell soma and processes both in culture and in brain slices.

How Do Specific Social Supports (Family, Friend, and Specialist) Reduce Stress in Patients With Substance Use Disorders: A Multiple Mediation Analysis.

Perceived social support has been found to reduce the stress of individuals who suffer from substance use disorders. However, the mediating effects of resilience and affect balance in the relationships between specific social supports (family, friend, and significant others) and perceived stress are still unclear. This study focused on substance use disorders (SUD) patients, exploring the mediating roles of resilience and affect balance on the relationships between three dimensions of social supports (family, friend, and specialist) and stress. Three hundred thirty-nine participants completed questionnaires of perceived social support, resilience, affect balance, and stress. After controlling resilience and affect balance, the results suggested the effects of perceived family and specialist supports on perceived stress were fully mediated, and the association between perceived friend support and perceived stress is partially mediated. The multiple mediation analysis showed resilience is significant in mediating the relationship between specific perceived supports in all models, while affect balance is only significant in mediating the relationship between specialist support and perceived stress. Implications for enriching current theoretical research and strategies for government and practitioners were also discussed.

Cocaine experience induces functional adaptations in astrocytes: Implications for synaptic plasticity in the nucleus accumbens shell.

Astrocytes have become established as an important regulator of neuronal activity in the brain. Accumulating literature demonstrates that cocaine self-administration in rodent models induces structural changes within astrocytes that may influence their interaction with the surrounding neurons. Here, we provide evidence that cocaine impacts astrocytes at the functional level and alters neuronal sensitivity to astrocyte-derived glutamate. We report that a 14-day period of short access to cocaine (2 h/day) decreases spontaneous astrocytic Ca2+ transients and precipitates changes in astrocyte network activity in the nucleus accumbens shell. This is accompanied by increased prevalence of slow inward currents, a physiological marker of neuronal activation by astrocytic glutamate, in a subset of medium spiny neurons. Within, but not outside, of this subset, we observe an increase in duration and frequency of N-methyl-D-aspartate (NMDA) receptor-mediated synaptic events. Additionally, we find that the link between synaptic NMDA receptor plasticity and neuron sensitivity to astrocytic glutamate is maintained independent of drug exposure and is observed in both cocaine and saline control animals. Imaging analyses of neuronal Ca2+ activity show no effect of cocaine self-administration on individual cells or on neuronal network activity in brain slices. Therefore, our data indicate that cocaine self-administration promotes astrocyte-specific functional changes that can be linked to increased glutamate-mediated coupling with principal neurons in the nucleus accumbens. Such coupling may be spatially restricted as it does not result in a broad impact on network structure of local neuronal circuits.

Peroxidase-mimicking Activity of PCN-222(Fe) for Colorimetric Sensing of Acetylcholinesterase Activity and Inhibition.

A colorimetric method for detection of acetylcholinesterase (AChE) activity and inhibition was developed using metal organic frameworks (i.e., PCN-222(Fe)) with peroxidase-like activity. The blue tetramethylbenzidine oxidized by PCN-222(Fe) fades due to the reduction by acetylthiocholine chloride produced from AChE catalysis. The detection method shows a linear range of 0.05 - 10 mU mL-1 and a detection limit of 0.03 mU mL-1 AChE. Average recoveries in serum samples varied from 93 to 115% and relative standard deviation (RSD) was lower than 4.9%.

Aggregation-induced emission enhancement of gold nanoclusters in metal-organic frameworks for highly sensitive fluorescent detection of bilirubin.

A fluorescence analysis method based on gold nanocluster (AuNC) and metal-organic framework (MOF) composite materials (AuNCs@ZIF-8) was established for highly sensitive detection of bilirubin (BR). First, AuNCs@ZIF-8 was successfully obtained by co-precipitation and displayed an aggregation-induced emission enhancement by the confinement effect of the MOFs (i.e., ZIF-8). The product showed approximately 7.0 times enhancement in the quantum yield and longer fluorescence lifetime from 2.29 µs to 11.51 µs compared with AuNCs. When BR combined with the metal node Zn2+ of ZIF-8, the skeleton of the composite was destroyed, leading to a great decrease in the fluorescence intensity by the transformation of the AuNCs from the aggregated state to dispersed state. The linear range for the detection of BR was 0.1-5.0 µM, with the limit of detection (LOD) of 0.07 µM (S/N = 3). The AuNCs@ZIF-8 exhibited a selective response toward BR within 5 min and detected BR in human serum. The long-wavelength emission by AuNCs avoided the interference of the complex biomatrix background fluorescence, indicating their great application prospects for clinical diagnosis.

Ratiometric fluorescence determination of alkaline phosphatase activity based on dual emission of bovine serum albumin-stabilized gold nanoclusters and the inner filter effect.

A novel and convenient method for the ratiometric fluorescence detection of alkaline phosphatase (ALP) activity was proposed based on dual emission of bovine serum albumin-templated gold nanoclusters (BSA-AuNCs) and the mechanism of the inner filter effect between BSA-AuNCs and p-nitrophenol (PNP). First, ALP catalyzed the hydrolysis of the substrate p-nitrophenyl phosphate (PNPP) to produce PNP. PNP effectively quenched the emission peak of BSA-AuNCs at 410 nm because of the overlap in absorbance feature of PNP and the fluorescence spectrum of BSA-AuNCs, and the peak at 650 nm was almost unaffected. Thus, a sensitive ratiometric method for detection of ALP activity was developed using the fluorescence intensity of BSA-AuNCs at 650 nm as a reference signal. ALP activity versus the ratio of fluorescence intensities at 410 and 650 nm showed good linearity between 0.2 and 5 mU mL-1 (R2 = 0.9931) and high sensitivity with a detection limit of 0.03 mU mL-1 (S/N = 3). The developed sensing method was successfully applied to investigate ALP inhibitors and detect ALP in serum samples.

How Resilience Promotes Mental Health of Patients With DSM-5 Substance Use Disorder? The Mediation Roles of Positive Affect, Self-Esteem, and Perceived Social Support.

Objectives: The existing studies found that resilience is a salient trait that can significantly affect people's psychological well-being with substance use disorders (SUDs). However, few studies examined how the mechanisms are connected between resilience and mental health among patients with Diagnostic and Statistical Manual of Mental Disorders-fifth edition SUD. This study investigated the mediation effects of positive affect, perceived social support, and self-esteem on the effect of resilience on perceived stress and life satisfaction in SUD patients. Design: A total of 415 patients diagnosed with Diagnostic and Statistical Manual of Mental Disorders-fifth edition SUD from the south of China joined the research. Outcome Measures: The study applied Connor-Davidson Resilience Scale, Positive and Negative Affect Scale, Multidimensional Scale of Perceived Social Support, Rosenberg Self-Esteem Scale, and Satisfaction with Life Scale to measure patients' resilience, positive affect, self-esteem, perceived social support, perceived stress, and life satisfaction. Results: Structural equation model analysis revealed that positive affect and self-esteem partially mediate the relationship between resilience and perceived stress. In contrast, positive affect and perceived social support partially mediate the relationship between resilience and life satisfaction. Conclusion: The findings provide insights for evidence-based substance abuse intervention that positive affect, self-esteem, and perceived social support can conditional the effects of resilience on promoting the mental health of SUD patients.

How parental socioeconomic status contribute to children's sports participation in China: A cross-sectional study.

Socioeconomic status (SES) is an important factor contributing to health inequality. This study aimed to investigate factors that predict junior school students' sports participation, identify the mechanisms underlying transmission of social resources and assess the mediating effects of classmate support and parental involvement on the relationship between parental SES and children's sports participation. 4829 males and 4536 females (mean age = 13.56 years, standard deviation = 0.686 years) participated in the study. Multivariate regression was adopted to analyze the determinants of junior school students' sports participation and multiple mediation analyses were used to analyze the hypothesized model. The results indicated that parental SES is significantly and directly correlated with junior school students' sports participation. In addition, parental SES has a significant indirect effect on sports participation through classmate support and parental involvement. Furthermore, the mediation effect of classmate support on the association between parental SES and sports participation is stronger than that of parental involvement. To promote and facilitate the participation of junior school students', strategies should be developed by government and social workers to strengthen classmate support and parental involvement.

APOE genotype-dependent pharmacogenetic responses to rapamycin for preventing Alzheimer's disease.

The ε4 allele of Apolipoprotein (APOE4) is the strongest genetic risk factor for Alzheimer's disease (AD), the most common form of dementia. Cognitively normal APOE4 carriers have developed amyloid beta (Aß) plaques and cerebrovascular, metabolic and structural deficits decades before showing the cognitive impairment. Interventions that can inhibit Aß retention and restore the brain functions to normal would be critical to prevent AD for the asymptomatic APOE4 carriers. A major goal of the study was to identify the potential usefulness of rapamycin (Rapa), a pharmacological intervention for extending longevity, for preventing AD in the mice that express human APOE4 gene and overexpress Aß (the E4FAD mice). Another goal of the study was to identify the potential pharmacogenetic differences in response to rapamycin between the E4FAD and E3FAD mice, the mice with human APOE ε3 allele. We used multi-modal MRI to measure in vivo cerebral blood flow (CBF), neurotransmitter levels, white matter integrity, water content, cerebrovascular reactivity (CVR) and somatosensory response; used behavioral assessments to determine cognitive function; used biochemistry assays to determine Aß retention and blood-brain barrier (BBB) functions; and used metabolomics to identify brain metabolic changes. We found that in the E4FAD mice, rapamycin normalized bodyweight, restored CBF (especially in female), BBB activity for Aß transport, neurotransmitter levels, neuronal integrity and free fatty acid level, and reduced Aß retention, which were not observe in the E3FAD-Rapa mice. In contrast, E3FAD-Rapa mice had lower CVR responses, lower anxiety and reduced glycolysis in the brain, which were not seen in the E4FAD-Rapa mice. Further, rapamycin appeared to normalize lipid-associated metabolism in the E4FAD mice, while slowed overall glucose-associated metabolism in the E3FAD mice. Finally, rapamycin enhanced overall water content, water diffusion in white matter, and spatial memory in both E3FAD and E4FAD mice, but did not impact the somatosensory responses under hindpaw stimulation. Our findings indicated that rapamycin was able to restore brain functions and reduce AD risk for young, asymptomatic E4FAD mice, and there were pharmacogenetic differences between the E3FAD and E4FAD mice. As the multi-modal MRI methods used in the study are readily to be used in humans and rapamycin is FDA-approved, our results may pave a way for future clinical testing of the pharmacogenetic responses in humans with different APOE alleles, and potentially using rapamycin to prevent AD for asymptomatic APOE4 carriers.

Long-Term Intranasal Insulin Aspart: A Profile of Gene Expression, Memory, and Insulin Receptors in Aged F344 Rats.

Intranasal insulin is a safe and effective method for ameliorating memory deficits associated with pathological brain aging. However, the impact of different formulations and the duration of treatment on insulin's efficacy and the cellular processes targeted by the treatment remain unclear. Here, we tested whether intranasal insulin aspart, a short-acting insulin formulation, could alleviate memory decline associated with aging and whether long-term treatment affected regulation of insulin receptors and other potential targets. Outcome variables included measures of spatial learning and memory, autoradiography and immunohistochemistry of the insulin receptor, and hippocampal microarray analyses. Aged Fischer 344 rats receiving long-term (3 months) intranasal insulin did not show significant memory enhancement on the Morris water maze task. Autoradiography results showed that long-term treatment reduced insulin binding in the thalamus but not the hippocampus. Results from hippocampal immunofluorescence revealed age-related decreases in insulin immunoreactivity that were partially offset by intranasal administration. Microarray analyses highlighted numerous insulin-sensitive genes, suggesting insulin aspart was able to enter the brain and alter hippocampal RNA expression patterns including those associated with tumor suppression. Our work provides insights into potential mechanisms of intranasal insulin and insulin resistance, and highlights the importance of treatment duration and the brain regions targeted.

The Relationship Between Self-Control and Self-Efficacy Among Patients With Substance Use Disorders: Resilience and Self-Esteem as Mediators.

Background: While substance use disorder is one of the overarching health and social issues that might seriously disrupt individuals' self-control and self-efficacy, most previous studies have been conducted among university students or other groups, and little is known about how the underlying mechanisms between self-control and self-efficacy might impact patients with substance use disorders. Objectives: The purpose of this study is to investigate how resilience and self-esteem mediate the relationships between self-control and self-efficacy among patients with substance use disorders. Methods: We conducted a cross-sectional study of 298 patients with substance use disorder from Shifosi rehab in China. Diagnostic and Statistical Manual of Mental Disorders (5th Edition)-based diagnostic questionnaires were used to collect demographic information and assess addiction severity. The Dual-Modes of Self-Control Scale (DMSC-S) was implemented to measure self-control, while self-esteem was measured using the Self-esteem Scale (SES). The Connor-Davidson Resilience Scale (CD-RISC) was used to measure resilience, and self-efficacy was measured by the regulatory emotional self-efficacy scale (RESE). Results: The correlations between all the dimensions and total scores on the self-control, resilience, self-esteem, and self-efficacy were significantly positive (p < 0.01), indicating that they could predict patients' self-efficacy. Bootstrap testing indicated that resilience and self-esteem fully mediated the relationship between self-control and self-efficacy, relationships between self-control and self-esteem were partially mediated by resilience, and resilience partially mediated the relationship between self-esteem and self-efficacy. Finally, the multiple-group analysis indicated that the relationships among self-control, resilience, self-esteem, and self-efficiency did not differ with respect to gender. Conclusions: The path from self-control through resilience and self-esteem and on to self-efficacy is significant among patients with substance use disorders, suggesting that increasing self-control, resilience, and self-esteem can improve self-efficacy among patients with substance use disorders.

Adsorption of cholesterol oxidase and entrapment of horseradish peroxidase in metal-organic frameworks for the colorimetric biosensing of cholesterol.

The co-immobilization of two enzymes onto single support commonly exhibits low efficiency due to the competition against limited sites. Water-stable metal-organic frameworks (MOFs) [i.e., PCN-333(Al)] with a high surface area and ultra-large cavities were employed to efficiently adsorb cholesterol oxidase (ChOx) and encapsulate horseradish peroxidase (HRP), respectively. The prepared PCN-333/ChOx&HRP was characterized through SEM, XRD, confocal microscopy, N2 adsorption isotherms, and thermal gravity analysis (TGA). The high surface area and high concentration of mesoporous cages resulted in the high loadings of both ChOx and HRP. The absorbed ChOx and the encapsulated HRP presented excellent activities without additional chemical modification. The immobilized enzymes were stable against protease digestion, organic solvents, temperature changes, and pH variation. Thus, a colorimetric biosensor for cholesterol detection was fabricated depending on cascade catalytic reactions of the immobilized bi-enzymes. An extended linear range from 0.0 to 40.0 µM with a low detection limit of 0.6 µM was obtained using the biosensor. The co-immobilization of the enzymes onto the surface and into the mesopores of MOFs provided a new and excellent platform for the development of highly stable and sensitive colorimetric biosensors.

The relationship among social support, self-esteem, affect balance and loneliness in individuals with substance use disorders in China.

This study analyzed the potential mediating role of self-esteem and affect balance on the relationship between social support and loneliness. Respondents were 426 substabce use disorders from the Shifosi and Dalianshan rehab facilities in China who had completed the Multidimensional Scale of Perceived Social Support, Rosenberg Self-Esteem Scale, Positive and Negative Affect Scales, and UCLA Loneliness Scale. The results indicated that self-esteem and affect balance fully mediated the relationship between perceived social support and loneliness and all the paths, ranging from social support through self-esteem and affect balance to loneliness, were significant. Finally, we analyzed possible approaches to decreasing individuals with substance use disorders' loneliness.

Social Support and Resilience as Mediators Between Stress and Life Satisfaction Among People With Substance Use Disorder in China.

This study investigated the potential mediating roles of resilience and social support in the relationship between stress and life satisfaction. A total of 426 individuals, who have substance use disorder, from the Shifosi and Dalianshan rehabilitation facilities in China participated in the study. They were tested using the Perceived Stress Scale, Multidimensional Scale of Perceived Social Support, Connor-Davidson Resilience Scale, and Satisfaction with Life Scale. Results showed that the serial multiple mediation of social support and resilience in the relationship between stress and life satisfaction was significant. Furthermore, the findings corroborate the important roles of perceived social support and resilience in alleviating stress. Finally, we discussed ways to enhance the life satisfaction for individuals who have substance use disorder and analyzed the limitations of this study.