Contextual Fear Learning and Extinction in the Primary Visual Cortex of Mice / 神经科学通报·英文版

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 Ca2+ 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 Ca2+ 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 Ca2+ 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.

Effect of different test indicators and observation periods in contextual fear conditioning test on detection of behavioral efficacy of 5XFAD transgenic mice / 上海交通大学学报（医学版）

Objective • To explore whether different test indicators and observation periods in contextual fear conditioning test affect the detection effectiveness of learning and memory ability of 5XFAD transgenic mice. Methods • Twelve 4-month-old female 5XFAD transgenic progeny mice and 14 4-month-old female LM progeny mice were selected, which were produced by crossing male 5XFAD transgenic mice and female C57BL/6 mice, to conduct open field test and contextual fear conditioning test in succession. Total distance and velocity in open field test and average motion index in the first 60 s of training stage in contextual fear conditioning test of the two groups of mice were used to evaluate the difference of locomotor activity. Besides, the first 180 s, 181-360 s and the first 300 s of testing stage for observation were selected to evaluate the selection effects on percent freeze and activity suppression ratio. Then further investigation was launched to explore the effects of different observation periods on the detection effectiveness of percent freeze and activity suppression ratio. Results • The differences of total distance and total velocity of the two groups of mice in open field test were not statistically significant, however the average motion index in the first 60 s of training stage in contextual fear conditioning test of 5XFAD transgenic mice was significantly higher than that of LM mice (P=0.027). The comparison of percent freeze among the three groups of observation periods of LM mice had significant difference (both P<0.05), while there was no statistical significance in activity suppression ratio. The comparisons of percent freeze and activity suppression ratio among the three groups of observation periods of 5XFAD mice had no significant difference. The differences of percent freeze between 5XFAD mice and LM mice was not statistically significant during the three observation periods. However, the activity suppression ratio of 5XFAD mice was significantly higher than that of LM mice in the first 180 s (P=0.038), in the other two observation periods the difference of activity suppression ratio between the two groups of mice was not statistically significant. Conclusion • The average motion index detected in training stage in contextual fear conditioning test is more sensitive than total distance and total velocity detected in open field test for evaluating locomotor activity of 5XFAD mice. In contextual fear conditioning test, different observation periods have effects on the value of percent freeze provided by single strain of mice, while the value of activity suppression ratio remains unaffected. Activity suppression ratio is more precise than percent freeze to reflect the cognitive deficiency of 5XFAD mice. It is more accurate to select the first 180 s of testing stage for observation.

Behavioral evaluation of male and female carioca high- and low-freezing rats / Avaliação comportamental de ratos machos e fêmeas cariocas / Evaluación comportamental em ratas machos y hembras cariocas alto y bajo congelamineto

Recent years have seen growing interest in the development of genetic animal models to investigate the bidirectional relationship between trait anxiety and defensive reactions. The present study further analyzed behavioral correlates of two novel breeding lines of rats, Carioca high-and low-conditioned freezing (CHF and CLF), based on defensive freezing responses to contextual cues previously associated with electric footshock. Male and female rats from the 10th generation were used to assess anxiety-like reactions in the elevated plus maze (EPM), depressive-like behavior in the forced swim test (FST), and aversive memory in the contextual fear conditioning paradigm. In the EPM, female rats showed lower anxiety-like behavior than males, whereas CHF rats were more anxious than CLF rats. The same pattern of results was found in the contextual fear conditioning paradigm. No differences were found between lines or sexes in the FST. Such differences in emotionality responses in the 10th generation of selected rats further indicate a possible use of this model to study correlations between trait anxiety and defensive reactions. The face validity of the model and its use to experimentally simulate generalized anxiety disorder in humans are also discussed.

Nos últimos anos, tem crescido o interesse pelo desenvolvimento de modelos animais geneticamente modificados para o estudo da relação bidirecional entre ansiedade traço e reações de defesa. O presente estudo analisou a correlação comportamental de duas novas linhagens de ratos, denominados de Carioca Alto e Baixo Congelamento (CHF e CLF), baseado na resposta de congelamento a estímulos contextuais previamente associados a choque elétrico nas patas. Foram utilizados ratos machos e fêmeas da 10ª geração para serem avaliadas as reações relacionadas à ansiedade no labirinto em cruz elevado (LCE), comportamentos relacionados à depressão no teste do nado forçado (TNF) e a memória aversiva no paradigma do medo condicionado ao contexto. No LCE, ratas fêmeas apresentaram um menor índice de comportamento indicativo de ansiedade comparadas aos machos. Mais ainda, animais da linhagem CHF apresentaram maiores índices de ansiedade em relação a animais da linhagem CLF. O mesmo padrão de resultados foi encontrado no paradigma do medo condicionado ao contexto. Não foram encontradas diferenças entre as linhagens ou entre os sexos testados no TNF. Tais diferenças nas respostas emocionais da 10ª geração de ratos geneticamente selecionados indicam uma possível utilização deste modelo no estudo de correlações entre ansiedade traço e reações de defesa. A validade de face do modelo e seu uso para simular experimentalmente o transtorno da ansiedade generalizada em humanos também são discutidos.

En los ultimos años, ha crescido el interés por el desarrollo de modelos animales geneticamente modificados para el estudio de la relación bidireccional entre ansiedad de rasgo y reaciones de defensa. El presente estudio analisó la correlación existente en el comportamiento de dos nuevas linajes de ratones, denominados de Carioca Alto y Bajo Congelamiento (CHF e CLF), basado en la respuesta de congelamiento a estimulos contextuales previamente asociados a choque elétrico en las patas. Fueron utilizados ratones machos y hembras de la 10ª generación para que fuesen evaluadas las reacciones relacionadas a la ansiedad en el laberinto en cruz elevado (LCE), comportamientos relacionados a la depresión en el teste de natación forzada (TNF) y la memória aversiva en el paradigma del miedo condicionado al contexto. En el LCE, las ratazanas presentaron un menor índice de comportamiento indicativo de ansiedad, se comparadas a los ratones machos. Animales de la linaje CHF presentaran mayores índices de ansiedad que los animales de la linaje hembras CLF. El mismo patrón de resultados fue encontrado em el paradigma del miedo condicionado al contexto. No se encontraron diferencias entre los linajes ó entre los sexos testados en el TNF. Tales diferencias en las respuestas emocionales de la 10ª generación de ratones geneticamente selecionados indican una posible utilización de ese modelo en el estudio de correlación entre ansiedade de rasgo y reacciones de defensa. La validad de contenido yaparente del modelo y sus utilización para simular experimentalmente el transtorno de la ansiedad generalizada en humanos, también son discutidos.

Sleep pattern and learning in knockdown mice with reduced cholinergic neurotransmission

Impaired cholinergic neurotransmission can affect memory formation and influence sleep-wake cycles (SWC). In the present study, we describe the SWC in mice with a deficient vesicular acetylcholine transporter (VAChT) system, previously characterized as presenting reduced acetylcholine release and cognitive and behavioral dysfunctions. Continuous, chronic ECoG and EMG recordings were used to evaluate the SWC pattern during light and dark phases in VAChT knockdown heterozygous (VAChT-KDHET, n=7) and wild-type (WT, n=7) mice. SWC were evaluated for sleep efficiency, total amount and mean duration of slow-wave, intermediate and paradoxical sleep, as well as the number of awakenings from sleep. After recording SWC, contextual fear-conditioning tests were used as an acetylcholine-dependent learning paradigm. The results showed that sleep efficiency in VAChT-KDHET animals was similar to that of WT mice, but that the SWC was more fragmented. Fragmentation was characterized by an increase in the number of awakenings, mainly during intermediate sleep. VAChT-KDHET animals performed poorly in the contextual fear-conditioning paradigm (mean freezing time: 34.4±3.1 and 44.5±3.3 s for WT and VAChT-KDHET animals, respectively), which was followed by a 45% reduction in the number of paradoxical sleep episodes after the training session. Taken together, the results show that reduced cholinergic transmission led to sleep fragmentation and learning impairment. We discuss the results on the basis of cholinergic plasticity and its relevance to sleep homeostasis. We suggest that VAChT-KDHET mice could be a useful model to test cholinergic drugs used to treat sleep dysfunction in neurodegenerative disorders.

Panic-like behaviors in Carioca High-and Low-conditioned Freezing rats

Panic disorder involves both recurrent unexpected panic attacks and persistent concern about having additional attacks. Electrical stimulation of the dorsal periaqueductal gray (dPAG) is an animal model of both panic attack and panic disorder, whereas contextual fear conditioning represents a model of anticipatory anxiety. Previous research indicated that anxiety has an inhibitory effect on panic attack-like behavior. However, still unclear is the role that anticipatory anxiety plays in panic disorder-like behaviors. This issue was investigated with two lines of animals selectively bred for high (Carioca High-Freezing) and low (Carioca Low-Freezing) freezing in response to contextual cues associated with footshock. The results suggest that although anticipatory anxiety might exert an inhibitory effect on the expression of panic attack, it might also facilitate the pathogenesis of panic disorder.

Effects of contextual fear conditioning and pentylenetetrazol on panic-like reactions induced by dorsal periaqueductal gray stimulation with N-methyl-D-aspartate

Electrical or chemical stimulation of the dorsal periaqueductal gray (DPAG) has been accepted as an animal model of panic attacks. This study investigates the influence of anticipatory anxiety in the occurrence of panic-like behavior induced by N-methyl-D-aspartate (NMDA) microinjection into the DPAG of rats. Behavioral (i.e., contextual fear conditioning) and pharmacological (i.e., pentylenetetrazol) manipulations were employed as animal models of anticipatory anxiety. In the first experiment, animals exposed to contextual cues that had been previously associated with electric footshocks through contextual fear conditioning were less likely than non-conditioned control animals to display defensive reactions such as running and jumping in response to microinjection of NMDA (0.3 µl of 15.0 µg/µl) into the DPAG. In the second experiment, rats were injected intraperitoneally with the anxiogenic drug pentylenetetrazol (PTZ, 15 mg/kg) 5 minutes before receiving intra-DPAG microinfusion with the same dose of NMDA as in Experiment 1. Panic-related behaviors were registered in an experimental arena immediately after NMDA microinfusion. As compared with saline pre-treated animals, PTZ significantly attenuated NMDA-induced panic-like reactions. These results further demonstrate the usefulness of DPAG chemical stimulation as an animal model of panic attacks and suggest that behavioral and pharmacological activation of the brain mechanisms underlying anticipatory anxiety might exert an antipanic-like effect.