Behavioral and Neurobiological Assessments of Predator-Based Fear Conditioning and Extinction

Shock, immobilization, and exposure to predator-related stimuli have all been used to study fear conditioning in rodents, but they have never been used in conjunction in a single study. Experiment 1 compared the effects of these three reinforcers, alone and in various combinations, on the expression of long-term conditioned fear memory and extinction in adult male rats. Whereas foot shock conditioning, alone, was rapidly extinguished;the combination of immobilization and cat exposure, or all 3 stimuli together, produced a significant increase in the magnitude of fear conditioning and greater resistance to extinction, which persisted for at least 5 weeks post-training (p < 0.05). Experiment 2 assessed the role of the hippocampus in predator-based context and cued fear conditioning. Pharmacological suppression of hippocampal activity during fear conditioning produced a selective impairment of contextual, but not cued, fear memory. Experiment 3 investigated the effects of sleep deprivation prior to fear conditioning on the expression of fear memory. This experiment demonstrated that pre-training sleep deprivation blocked the expression of contextual (hippocampal-dependent), but not cued (hippocampal-independent), fear memory. Overall, this series of experiments has extended the use of predator exposure in conjunction with conventional reinforcers, such as foot shock and immobilization, to advance our understanding of the neurobiology of traumatic memory.

Effects of Abnormal Early Rearing Environments on Fear Memory in Adult Rats

In the present experiment, Pavlovian fear conditioning was adopted to study the effects of different early rearing environments on fear conditioning in adult rats. Weaned rats were reared in three manipulable environments （enriched, social and isolated conditions）. After 8 weeks, fear conditioning （characterized by percentage of freezing） was observed and analyzed, and rats＇ weight, locomoter activity and foot-shock sensitivity were operated too. The results showed that： （1） Compared with control group, the level of conditioned fear was significantly increased in enriched group, but significantly decreased in isolated group; （2） Enriched and isolated conditions influenced rat＇s weight significantly; （3） Different rearing conditions have no effect on locomoter activity and foot-shock sensitivity. These results indicated that early enriched condition could improve the tone-evoked fear conditioning response, while isolated condition impaired the response.

GLYX-13 pretreatment ameliorates long-term isoflurane exposure-induced cognitive impairment in mice

Accumulating evidence indicates that inhalation anesthetics induce or increase the risk of cognitive impairment. GLYX-13（rapastinel） acts on the glycine site of N-methyl-D-aspartate receptors（NMDARs） and has been shown to enhance hippocampus-dependent learning and memory function. However, the mechanisms by which GLYX-13 affects learning and memory function are still unclear. In this study, we investigated these mechanisms in a mouse model of long-term anesthesia exposure. Mice were intravenously administered 1 mg/kg GLYX-13 at 2 hours before isoflurane exposure（1.5% for 6 hours）. Cognitive function was assessed using the contextual fear conditioning test and the novel object recognition test. The mRNA expression and phosphorylated protein levels of NMDAR pathway components, N-methyl-D-aspartate receptor subunit 2B（NR2B）-Ca2+/calmodulin dependent protein kinase II（CaMKII）-cyclic adenosine monophosphate response element binding protein（CREB）, in the hippocampus were evaluated by quantitative RT-PCR and western blot assay. Pretreatment with GLYX-13 ameliorated isoflurane exposure-induced cognitive impairment and restored NR2B, CaMKII and CREB mRNA and phosphorylated protein levels. Intracerebroventricular injection of KN93, a selective CaMKII inhibitor, significantly diminished the effect of GLYX-13 on cognitive function and NR2B, CaMKII and CREB levels in the hippocampus. Taken together, our findings suggest that GLYX-13 pretreatment alleviates isoflurane-induced cognitive dysfunction by protecting against perturbation of the NR2B/CaMKII/CREB signaling pathway in the hippocampus. Therefore, GLYX-13 may have therapeutic potential for the treatment of anesthesia-induced cognitive dysfunction. This study was approved by the Experimental Animal Ethics Committee of Drum Tower Hospital affiliated to the Medical College of Nanjing University, China（approval No. 20171102） on November 20, 2017.

Endothelin-1-induced mini-stroke in the dorsal hippocampus or lateral amygdala results in deficits in learning and memory

Functional and structural alterations in brain connectivity associated with brain ischemia have been extensively studied. However, the mechanism whereby local ischemia in deep brain region affect brain functions is still unknown. Here, we first established a mini-stroke model by infusion of endothelin-1 （ET-1） into the dorsal hippo- campus or the lateral amygdala, and then investigated how these mini-infarcts affected brain functions associated with these regions. We found that rats with ET-1 infusion showed deficit in recall of contextual fear memory, but not in learning process and recall of tone fear memory. In novel object task, ET-1 in the hippocampus also elimi- nated object identity memory. ET-1 in the lateral amygdale affected acquisition of fear conditioning and disrupted retention of tone-conditioned fear, but did not impair retention of contextual fear. These findings suggest that ET-1- induced mini-infarct in deep brain area leads to functional deficits in learning and memory associated with these regions.

Female Carriers of the Met Allele of the BDNF Val66Met Polymorphism Develop Weaker Fear Memories in a Fear-Potentiated Startle Paradigm

The val66met polymorphism of the bdnf gene, which is associated with compromised brain-derived neurotrophic factor (BDNF) signaling, impaired synaptic plasticity, and impaired learning, may increase one’s susceptibility to stress- and anxiety-related disorders. Indeed, previous work has reported greater anxiety-related behaviors and impairments of fear conditioning and extinction in individuals who carry the met allele that results from this polymorphism. Nevertheless, findings in this area of research have been equivocal. Thus, we examined the influence of the val66met polymorphism on fear conditioning, extinction, and extinction memory testing. One hundred and twenty healthy participants completed differential fear conditioning in a fear-potentiated startle paradigm, followed by extinction and extinction memory testing 24 and 48 hr later, respectively. Participants were genotyped for the val66met polymorphism and divided into met allele carriers and non-carriers. Results revealed that, although both met-carriers and non-carriers developed conditioned fear, met-carriers exhibited significantly weaker fear acquisition than non-carriers. This difference persisted throughout extinction and extinction memory testing and, during these last two days of testing, was primarily evident in females. These results are consistent with previous work demonstrating that this polymorphism is associated with impaired amygdala-dependent fear learning and extend such findings by demonstrating that females may be more sensitive to such effects.

Contrasting the Amnesic Effects of Temporary Inactivation with Lesions of the Hippocampus on Context Memory

Lesions and temporary inactivation of the hippocampus (HPC) in rodents occasionally lead to discrepant amnesic effects. We directly compared and contrasted the retrograde amnesic effects that small HPC lesions (~50% damage), large HPC lesions (~80% damage), and combined dorsal and ventral HPC inactivation using the sodium channel blocker tetrodotoxin (TTX) had on contextual fear conditioning. Compared to control rats, large HPC lesions significantly reduced freezing during retention testing, a behaviour consistent with retrograde amnesia. In contrast, neither the small lesions nor the TTX inactivation significantly reduced freezing. The extent of damage was significantly and negatively correlated with retention performance (r(9) = -0.896, p < 0.001), suggesting that 70% or more of the HPC needed to be damaged to observe deficits. Importantly, TTX inactivation disrupted spatial memory in the Morris Water Task, confirming that our inactivation procedure did impair one form of HPC-dependent memory. To assess the extent of the TTX inactivation, immediate early gene expression was quantified in the HPC following the Morris Water Task. However, despite the behavioural impairment, we did not find a significant reduction in expression. We conclude that temporary inactivation of the HPC may fail to impair context fear memory because this technique does not sufficiently disrupt the HPC.

A Study of Transcranial magnetic stimulation For Extinction about Early Traumatic Memory Description

Individuals will experience strong traumatic memories after traumatic events such as earthquakes,car accidents,or loss of loved ones,the most common of which is fear memory.In this experiment,a multisensory compound stimulation model(electric stimulation+picture)was used as the conditional stimulus,and the skin electrical response was used as the index of fear response.The effect of retrievalextinction paradigm on the conditional fear extinction was tested.In addition,high-frequency rTMS was introduced as a treatment.To explore the therapeutic effect of TMS on conditional fear,with the aim of providing new measures for the treatment of PTSD.

Establishment of Visual Fear Conditioning in Long-evans Rats

The neural mechanisms underlying visual information transmission and coding are currently attracting the attention of neuroscience and brain-like computing scholars.The subcortical visual pathway is known to affect fear emotion regulation via the amygdala;however an experimental paradigm for visual fear cognition training remains undefined.In this study,Long-Evans(LE)rats were used to develop an experimental training paradigm for visual cognition-associated fear conditioning based on the Pavlovian conditioning reflex.Simple images were shown on a unilateral screen(conditioned stimulus)were combined with electric foot shocks(unconditioned stimulus).We designed training paradigms and set up an estimated index using the rate of successful active escape.The training results were analyzed using a two-way ANOVA,and curve fitting was used to analyze the influence of decision time between the conditioned stimulus(CS)and unconditioned stimulus(US)on choice behavior.While neither the CS nor US had a significant effect on visual fear association training in LE rats,the decision time duration(CS-US)did have an impact on training.The method described here is most effective in establishing visual fear associations in rats when the(CS-UC)=10 s.This study describes a new experimental training paradigm for fear conditioning using visual stimuli and a quantitative evaluation standard according to the training mode of visual stimulation graphics.Moreover,it is an efficient paradigm for future study on visual information-processing mechanisms in the subcortical visual pathway during fear conditioning.

Contextual Fear Learning and Extinction in the Primary Visual Cortex of Mice

Fear memory contextualization is critical for selecting adaptive behavior to survive.Contextual fear conditioning(CFC)is a classical model for elucidating related underlying neuronal circuits.The primary visual cortex(V1)is the primary cortical region for contextual visual inputs,but its role in CFC is poorly understood.Here,our experiments demonstrated that bilateral inactivation of V1 in mice impaired CFC retrieval,and both CFC learning and extinction increased the turnover rate of axonal boutons in V1.The frequency of neuronal Ca^(2+)activity decreased after CFC learning,while CFC extinction reversed the decrease and raised it to the naïve level.Contrary to control mice,the frequency of neuronal Ca^(2+)activity increased after CFC learning in microglia-depleted mice and was maintained after CFC extinction,indicating that microglial depletion alters CFC learning and the frequency response pattern of extinction-induced Ca^(2+)activity.These findings reveal a critical role of microglia in neocortical information processing in V1,and suggest potential approaches for cellular-based manipulation of acquired fear memory.

On brain activity mapping: Insights and lessons from Brain Decoding Project to map memory patterns in the hippocampus

The BRAIN project recently announced by the president Obama is the reflection of unrelenting human quest for cracking the brain code, the patterns of neuronal activity that define who we are and what we are. While the Brain Activity Mapping proposal has rightly emphasized on the need to develop new technologies for measuring every spike from every neuron, it might be helpful to consider both the theoretical and experimental aspects that would accelerate our search for the organizing principles of the brain code. Here we share several insights and lessons from the similar proposal, namely, Brain Decoding Project that we initiated since 2007. We provide a specific example in our initial mapping of real-time memory traces from one part of the memory circuit, namely, the CA1 region of the mouse hippocampus. We show how innovative behavioral tasks and appropriate mathematical analyses of large datasets can play equally, if not more, important roles in uncovering the specific-to-general feature-coding cell assembly mechanism by which episodic memory, semantic knowledge, and imagination are generated and organized. Our own experiences suggest that the bottleneck of the Brain Project is not only at merely developing additional new technologies, but also the lack of efficient avenues to disseminate cutting edge platforms and decoding expertise to neuroscience community. Therefore, we propose that in order to harness unique insights and extensive knowledge from various investigators working in diverse neuroscience subfields, ranging from perception and emotion to memory and social behaviors, the BRAIN project should create a set of International and National Brain Decoding Centers at which cutting-edge recording technologies and expertise on analyzing large datasets analyses can be made readily available to the entire community of neuroscientists who can apply and schedule to perform cutting-edge research.