小鼠动物实验方法系列专题(五) 应用高架十字迷宫分析小鼠焦虑行为

高架十字迷宫(the elevated plus maze,EPM)是研究小鼠焦虑行为的重要实验,本文介绍了EPM的原理和实验步骤。将小鼠置于开臂闭臂接合处,面向开臂,通过录像记录分析小鼠在EPM的表现。结果发现,两种小鼠均可成功完成实验,C57小鼠在EPM内较不活跃,闭臂滞留时间占总时间百分比显著高于129Sv小鼠。应用EPM可以简单直观地分析小鼠焦虑行为。     

小鼠动物实验方法系列专题(七)旷场实验在小鼠行为分析中的应用

旷场实验是一个研究小鼠自发活动与探索行为的实验。该文以GAT1基因敲除小鼠为例介绍了旷场实验的原理和实验步骤。将小鼠置于箱型的旷场装置中,通过录像记录并分析小鼠在旷场中的活动。结果发现,GAT1基因敲除小鼠的自发活动与野生型小鼠无明显差异,而GAT1敲除小鼠的趋避性及焦虑水平较低。旷场实验简单、有效,是一项重要的行为学实验。     

Study of temporomandibular joint disorder in older patients by magnetic resonance imaging

doi: 10.1111/j.1741‐2358.2011.00552.x Study of temporomandibular joint disorder in older patients by magnetic resonance imaging (MRI) Objectives: To compare characteristics in older patients in a sample of the general population of those with temporomandibular joint disorder (TMJD). Materials and methods: A prospective study was carried out between 2001 and 2008 in patients with TMJD. The whole sample consisted of 141 patients divided in two groups: 31 patients aged over 60 (median age 67.9, ranging from 60 to 82) and the remaining 110 patients (median age 36.3, ranging from 12 to 59) who were seeking treatment. Clinical diagnostics was confirmed by MRI. Pain intensity was rated on a visual analogue scale (VAS 0‐10). Results: There was no statistical difference between average pain in older patients (6.2) and patients aged up to 59 (5.7) evaluated by VAS. There was a statistically significant difference (p = 0.002) in pain duration: older patients reported shorter duration of experienced pain (7.8 months) than patients aged up to 59 (12.2 months). Conclusion: In this study, it was found that 22% were older patients with TMJD. A higher level of anxiety was shown in both patients’ groups, regardless of shorter pain experience in the older patients.     

Hippocampal SPARC regulates depression‐related behavior

SPARC (secreted protein acidic and rich in cysteine) is a matricellular protein highly expressed during development, reorganization and tissue repair. In the central nervous system, glial cells express SPARC during development and in neurogenic regions of the adult brain. Astrocytes control the glutamate receptor levels in the developing hippocampus through SPARC secretion. To further characterize the role of SPARC in the brain, we analyzed the hippocampal‐dependent adult behavior of SPARC KO mice. We found that SPARC KO mice show increased levels of anxiety‐related behaviors and reduced levels of depression‐related behaviors. The antidepressant‐like phenotype could be rescued by adenoviral vector‐mediated expression of SPARC in the adult hippocampus, but anxiety‐related behavior persisted in these mice. To identify the cellular mechanisms underlying these behavioral alterations, we analyzed neuronal activity and neurogenesis in the dentate gyrus (DG). SPARC KO mice have increased levels of neuronal activity, evidenced as more neurons that express c‐Fos after a footshock. SPARC also affects cell proliferation in the subgranular zone of the DG, although it does not affect maturation and survival of new neurons. SPARC expression in the adult DG does not revert the proliferation phenotype in KO mice, but our results suggest a role of SPARC in limiting the survival of new neurons in the DG. This work suggests that SPARC could affect anxiety‐related behavior by modulating neuronal activity, and that depression‐related behavior is dependent upon the adult expression of SPARC, which affects adult brain function by mechanisms that need to be elucidated.     

Real-time fMRI Biofeedback Targeting the Orbitofrontal Cortex for Contamination Anxiety

We present a method for training subjects to control activity in a region of their orbitofrontal cortex associated with contamination anxiety using biofeedback of real-time functional magnetic resonance imaging (rt-fMRI) data. Increased activity of this region is seen in relationship with contamination anxiety both in control subjects¹ and in individuals with obsessive-compulsive disorder (OCD),² a relatively common and often debilitating psychiatric disorder involving contamination anxiety. Although many brain regions have been implicated in OCD, abnormality in the orbitofrontal cortex (OFC) is one of the most consistent findings.^{3, 4} Furthermore, hyperactivity in the OFC has been found to correlate with OCD symptom severity⁵ and decreases in hyperactivity in this region have been reported to correlate with decreased symptom severity.⁶ Therefore, the ability to control this brain area may translate into clinical improvements in obsessive-compulsive symptoms including contamination anxiety. Biofeedback of rt-fMRI data is a new technique in which the temporal pattern of activity in a specific region (or associated with a specific distributed pattern of brain activity) in a subject''s brain is provided as a feedback signal to the subject. Recent reports indicate that people are able to develop control over the activity of specific brain areas when provided with rt-fMRI biofeedback.^7-12 In particular, several studies using this technique to target brain areas involved in emotion processing have reported success in training subjects to control these regions.^13-18 In several cases, rt-fMRI biofeedback training has been reported to induce cognitive, emotional, or clinical changes in subjects.^{8, 9, 13, 19} Here we illustrate this technique as applied to the treatment of contamination anxiety in healthy subjects. This biofeedback intervention will be a valuable basic research tool: it allows researchers to perturb brain function, measure the resulting changes in brain dynamics and relate those to changes in contamination anxiety or other behavioral measures. In addition, the establishment of this method serves as a first step towards the investigation of fMRI-based biofeedback as a therapeutic intervention for OCD. Given that approximately a quarter of patients with OCD receive little benefit from the currently available forms of treatment,^20-22 and that those who do benefit rarely recover completely, new approaches for treating this population are urgently needed.     

Adenylyl cyclase-5 activity in the nucleus accumbens regulates anxiety-related behavior

Type 5 adenylyl cyclase (AC5) is highly concentrated in the dorsal striatum and nucleus accumbens (NAc), two brain areas which have been implicated in motor function, reward, and emotion. Here we demonstrate that mice lacking AC5 (AC5-/-) display strong reductions in anxiety-like behavior in several paradigms. This anxiolytic behavior in AC5-/- mice was reduced by the D(1) receptor antagonist SCH23390 and enhanced by the D(1) dopamine receptor agonist, dihydrexidine (DHX). DHX-stimulated c-fos induction in AC5-/- mice was blunted in the dorso-lateral striatum, but it was overactivated in the dorso-medial striatum and NAc. The siRNA-mediated inhibition of AC5 levels within the NAc was sufficient to produce an anxiolytic-like response. Microarray and RT-PCR analyses revealed an up-regulation of prodynorphin and down-regulation of cholecystokinin (CCK) in the NAc of AC5-/- mice. Administration of nor-binaltorphimine (a kappa opioid receptor antagonist) or CCK-8s (a CCK receptor agonist) reversed the anxiolytic-like behavior exhibited by AC5-/- mutants. Taken together, these results suggest an essential role of AC5 in the NAc for maintaining normal levels of anxiety.     

Artichoke leaf extract reduces mild dyspepsia in an open study

A recent post-marketing study indicated that high doses of standardised artichoke leaf extract (ALE) may reduce symptoms of dyspepsia. To substantial these findings, this study investigated the efficacy of a low-dose ALE on amelioration of dyspeptic symptoms and improvement of quality of life. The study was an open, dose-ranging postal study. Healthy patients with self-reported dyspepsia were recruited through the media. The Nepean Dyspepsia Index and the State-Trait Anxiety Inventory were completed at baseline and after 2 months of treatment with ALE, which was randomly allocated to volunteers as 320 or 640 mg daily. Of the 516 participants, 454 completed the study. In both dosage groups, compared with baseline, there was a significant reduction of all dyspeptic symptoms, with an average reduction of 40% in global dyspepsia score. However, there were no differences in the primary outcome measures between the two groups, although relief of state anxiety, a secondary outcome, was greater with the higher dosage (P = 0.03). Health-related quality of life was significantly improved in both groups compared with baseline. We conclude that ALE shows promise to ameliorate upper gastro-intestinal symptoms and improve quality of life in otherwise healthy subjects suffering from dyspepsia.     

道路交通噪声对金色中仓鼠焦虑行为及应激生理的影响

人类活动产生的噪声污染对动物和人类的影响正受到日益增多的关注。本文以雄性成年金色中仓鼠为实验动物模型,探讨了北京市主干道交通噪声对其焦虑行为及血象、应激生理的影响。分别以北京主干道噪声（80±10 dB SPL）暴露为实验组,实验室环境噪声（50±4 dB SPL）暴露为对照组,噪声处理动物1小时后进行旷场行为学测试,然后取血对比观测两组鼠血液学指标和应激响应、抗氧化酶活性等生理指标的变化。结果显示道路交通噪声没有导致仓鼠出现明显的焦虑行为;不过,实验组血小板数显著低于对照组(P = 0.044),其他血象指标两组间差异不显著;噪声对血清皮质醇,谷丙、谷草转氨酶影响不显著;实验组的血清谷胱甘肽过氧化物酶（GSH-Px）活性极显著低于对照组（P < 0.001）,但超氧化物歧化酶（SOD）活性、过氧化氢酶（CAT）活性、总抗氧化能力 (TAOC) 和丙二醛（MDA）水平两组间差异不显著;血清溶菌酶活性实验组降低较明显,接近显著水平（P = 0.0507）。我们的结果显示道路交通噪声胁迫导致了金色中仓鼠血象指标发生了变化,这提示北京市主干道交通噪声刺激对金色中仓鼠生理功能产生了一定的副作用。     

Disruption of behavior and brain metabolism in artificially reared rats

Early adverse life stress has been associated to behavioral disorders that can manifest as inappropriate or aggressive responses to social challenges. In this study, we analyzed the effects of artificial rearing on the open field and burial behavioral tests and on GFAP, c‐Fos immunoreactivity, and glucose metabolism measured in anxiety‐related brain areas. Artificial rearing of male rats was performed by supplying artificial milk through a cheek cannula and tactile stimulation, mimicking the mother's licking to rat pups from the fourth postnatal day until weaning. Tactile stimulation was applied twice a day, at morning and at night, by means of a camel brush on the rat anogenital area. As compared to mother reared rats, greater aggressiveness, and boldness, stereotyped behavior (burial conduct) was observed in artificially reared rats which occurred in parallel to a reduction of GFAP immunoreactivity in somatosensory cortex, c‐Fos immunoreactivity at the amygdala and primary somatosensory cortex, and lower metabolism in amygdala (as measured by 2‐deoxi‐2‐[¹⁸fluoro]‐d ‐glucose uptake, assessed by microPET imaging). These results could suggest that tactile and/or chemical stimuli from the mother and littermates carry relevant information for the proper development of the central nervous system, particularly in brain areas involved with emotions and social relationships of the rat. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1413–1429, 2017     

Role of the MT1 and MT2 melatonin receptors in mediating depressive‐ and anxiety‐like behaviors in C3H/HeN mice

Melatonin is a neurohormone primarily synthesized by the pineal gland following a circadian rhythm with a high level during the night and a low level during the day. Alterations in the synthesis and secretion of melatonin have been reported in various mood disorders, including major depressive disorder. However, the role of endogenous melatonin in the pathophysiology of depressive disorder is unclear. Melatonin primarily acts through two G protein‐coupled receptors, termed MT₁ and MT₂. The present study investigated the effect of genetic deletion of the MT₁ and/or MT₂ receptors on tests associated with depression‐ and anxiety‐like behaviors in C3H/HeN mice. Deletion of the MT₁ and/or MT₂ receptors caused a deficit in hedonic and social interaction behavior, and increased anxiety‐like behavior. It is likely that dysregulations of the MT₁ and/or MT₂ melatonin receptors could be involved in the pathophysiology of depression and anxiety.