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Itch is a cutaneous sensation that is critical in driving scratching behavior. The long-standing question of whether there are specific neurons for itch modulation inside the brain remains unanswered. Here, we report a subpopulation of itch-specific neurons in the ventrolateral orbital cortex (VLO) that is distinct from the pain-related neurons. Using a Tet-Off cellular labeling system, we showed that local inhibition or activation of these itch-specific neurons in the VLO significantly suppressed or enhanced itch-induced scratching, respectively, whereas the intervention did not significantly affect pain. Conversely, suppression or activation of pain-specific neurons in the VLO significantly affected pain but not itch. Moreover, fiber photometry and immunofluorescence verified that these itch- and pain-specific neurons are distinct in their functional activity and histological location. In addition, the downstream targets of itch- and pain-specific neurons were different. Together, the present study uncovers an important subpopulation of neurons in the VLO that specifically modulates itch processing.

Itch is a complex and unpleasant sensory experience. Recent studies have begun to investigate the neural mechanisms underlying the modulation of sensory and emotional components of itch in the brain. However, the key brain regions and neural mechanism involved in modulating the attentional processing of itch remain elusive. Here, we showed that the prelimbic cortex (PrL) is associated with itch processing and that the manipulation of itch-responsive neurons in the PrL significantly disrupted itch-induced scratching. Interestingly, we found that increasing attentional bias toward a distracting stimulus could disturb itch processing. We also demonstrated the existence of a population of attention-related neurons in the PrL that drive attentional bias to regulate itch processing. Importantly, itch-responsive neurons and attention-related neurons significantly overlapped in the PrL and were mutually interchangeable in the regulation of itch processing at the cellular activity level. Our results revealed that the PrL regulates itch processing by controlling attentional bias.

Itch is an unpleasant sensation that induces the desire to scratch. Except for a sketchy map focusing on neural mechanisms underlying itch processing being drawn at the peripheral and spinal level over the past decades, the brain mechanisms remain poorly understood. Several previous studies indicated that anterior cingulate cortex (ACC) and prelimbic cortex (PrL), two subregions of the medial prefrontal cortex (mPFC) play an important role in regulating itch processing. However, the knowledge about whether infralimbic cortex (IL), another subregion of mPFC, is involved in modulating itch processing remains unclear. Here, we showed that the activity of IL excitatory pyramidal neurons was significantly elevated during itch-related scratching, and pharmacogenetic inhibition of IL pyramidal neurons significantly impaired itch-related scratching. Moreover, IL-medial striatum (MS) projections were verified as a critical neural pathway for modulating itch processing. Therefore, the present study firstly presents the regulatory function of IL pyramidal neurons during itch processing and also reveals that IL-MS projections are involved in modulating the itch processing.

Although spinal cord injury is the main cause of disability and death worldwide, there is still no definite and effective treatment method for this condition. Our previous clinical trials confirmed that the increased excitability of the motor cortex was significantly related to the functional prognosis of patients with spinal cord injury. However, it remains unclear which cell types in the motor cortex lead to the later functional recovery and the neural mechanism of recovery. Herein, we used optogenetic technology to selectively activate glutamate neurons in the primary motor cortex and study whether activation of glutamate neurons in the primary motor cortex can promote functional recovery after spinal cord injury in rats and the preliminary neural mechanisms involved. Our results showed that the activation of glutamate neurons in the motor cortex can significantly improve the motor function score in rats and can effectively shorten the incubation period of motor evoked potentials and increase the amplitude of motor potentials. In addition, HE staining and nerve fiber staining at the injured site showed that accurate activation of the primary motor cortex could effectively promote tissue recovery and neurofilament growth (GAP-43, NF) at the injured site of the spinal cord, while the content of some growth-related proteins (BDNF, NGF) at the injured site increased. These results suggested that selective activation of glutamate neurons in the primary motor cortex can promote functional recovery after spinal cord injury and may be of great significance for understanding the neural cell mechanism underlying functional recovery induced by motor cortex stimulation.

Itch is an unpleasant sensation that evokes a desire to scratch. Itch processing in the peripheral and spinal cord has been studied extensively, but the mechanism of itch in the central nervous system is still unclear. Anterior cingulate cortex (ACC) and prelimbic cortex (Prl), two subregions of the prefrontal cortex closely related to emotion and motivation, have been reported to be activated during itching in a series of functional imaging studies. However, the exact role of Prl and the differences between ACC and Prl in itch modulation remains unknown. To directly test the differential roles of ACC and Prl in itch processing, we chemogeneticlly inhibited the caudal ACC and Prl, respectively. We found that inhibition of caudal ACC reduced histaminergic but not non-histaminergic itch-induced scratching behaviors. In contrast, inhibition of Prl reduced both histaminergic and non-histaminergic itch-induced scratching behaviors. Our study provided direct evidence of Prl involvement in itch modulation and revealed the differential roles of caudal ACC and Prl in regulating histaminergic and non-histaminergic itch.

Primary cerebellar agenesis (PCA), a brain disease where the cerebellum does not develop, is an extremely rare congenital disease with only eleven living cases reported thus far. Studies of the PCA case will thus provide valuable insights into the necessity of cerebellar development for controlling and modulating cognitive functions of the brain. In this follow-up study, we further investigated the performance of associative learning and time perception of a 26-year-old female complete PCA case. We assessed whether delayed eyeblink conditioning (EBC), which represents prototypical associative motor learning function of the cerebellum, could be partially compensated by the extracerebellar brain regions in complete absence of the cerebellum. We also assessed whether the cerebellum, a critical brain region for millisecond-range interval timing, is essential for perception of the second-range time interval. Twelve neurotypical age-matched individuals were used as controls. We found that although the complete PCA patient had only mild to moderate motor deficits, she was unable to perform the delayed EBC even after 1-week of extensive training. Additionally, the PCA patient also performed poorly during time reproduction experiments in which she overproduced the millisecond-range time intervals, while underproduced the second-range time intervals. The PCA patient also failed to perform the temporal eyeblink conditioning with a 5 s fixed interval as the conditioned stimulus. These results indicate that the cerebellum is indispensable for associative motor learning and involved in timing of sub-second intervals, as well as in the perception of second-range intervals.

The medial prefrontal cortex (mPFC) is closely involved in many higher-order cognitive functions, including learning to associate temporally discontiguous events (called temporal associative learning). However, direct evidence for the role of mPFC and the neural pathway underlying modulation of temporal associative motor learning is sparse. Here, we show that optogenetic inhibition of the mPFC or its axon terminals at the pontine nuclei (PN) during trace intervals or whole trial period significantly impaired the trace eyeblink conditioning (TEC), but had no significant effects on TEC during the conditioned stimulus or intertrial interval period. Our results suggest that activities associated with the mPFC-PN projection during trace intervals is crucial for trace associative motor learning. This finding is of great importance in understanding the mechanisms and the relevant neural pathways underlying mPFC modulation of temporal associative motor learning.

Diverse and powerful mechanisms have evolved to enable organisms to modulate learning and memory under a variety of survival conditions. Cumulative evidence has shown that the prefrontal cortex (PFC) is closely involved in many higher-order cognitive functions. However, when and how the medial PFC (mPFC) modulates associative motor learning remains largely unknown. Here, we show that delay eyeblink conditioning (DEC) with the weak conditioned stimulus (wCS) but not the strong CS (sCS) elicited a significant increase in the levels of c-Fos expression in caudal mPFC. Both optogenetic inhibition and activation of the bilateral caudal mPFC, or its axon terminals at the pontine nucleus (PN) contralateral to the training eye, significantly impaired the acquisition, recent and remote retrieval of DEC with the wCS but not the sCS. However, direct optogenetic activation of the contralateral PN had no significant effect on the acquisition, recent and remote retrieval of DEC. These results are of great importance in understanding the elusive role of the mPFC and its projection to PN in subserving the associative motor learning under suboptimal learning cue.

Prepulse inhibition (PPI) and habituation of the acoustic startle response (ASR) are considered to be effective neurobiological measures of sensorimotor gating and information processing. The deficit of PPI and habituation of ASR has been proposed to be candidate endophenotypes of schizophrenia spectrum disorders. However, there has been little information on PPI and ASR measures in Chinese. The present study aimed to provide more information about the characteristics of PPI and ASR in young healthy Chinese and investigate their sensitivity to experimental parameters and characteristics of population. In this study, we examined the PPI and habituation of ASR in 41 young healthy adults (21 males and 20 females), using an acoustic startle stimulus of 115 dB and a prepulse of 75 dB at a lead interval (LI) of 60 ms and 120 ms, respectively. The behavioral performance demonstrated that the PPI and habituation of ASR in all the young participants were robust. The significant difference was not observed in PPI and habituation between male and female. The block effect on PPI was significant; PPI reduces with increasing training. Latency facilitation was observed under prepulse conditions, with a significant effect of LI. Compared to previous studies in Caucasians, Chinese in this study shows a higher habituation and PPI. In conclusion, this research provides more data of behavioral characteristics of PPI and ASR in young healthy Chinese. Chinese in this study shows a higher habituation and PPI than Caucasians in previous studies.

It has been well established that the cerebellum and its associated circuitry constitute the essential neuronal system for both delay and trace classical eyeblink conditioning (DEC and TEC). However, whether the cerebellum is sufficient to independently modulate the DEC, and TEC with a shorter trace interval remained controversial. Here, we used direct optogenetic stimulation of mossy fibers in the middle cerebellar peduncle (MCP) as a conditioned stimulus (CS) replacement for the peripheral CS (eg, a tone CS or a light CS) paired with a periorbital shock unconditioned stimulus (US) to examine the ability of the cerebellum to learn the DEC and the TEC with various trace intervals. Moreover, neural inputs to the pontine nucleus (PN) were pharmacological blocked to limit the associative motor learning inside the cerebellum. We show that all rats quickly acquired the DEC, indicating that direct optogenetic stimulation of mossy fibers in the left MCP is a very effective and sufficient CS to establish DEC and to limit the motor learning process inside the cerebellum. However, only five out of seven rats acquired the TEC with a 150-ms trace interval, three out of nine rats acquired the TEC with a 350-ms trace interval, and none of the rats acquired the TEC with a 500-ms trace interval. Moreover, pharmacological blocking glutamatergic and GABAergic inputs to the PN from the extra-cerebellar and cerebellar regions has no significant effect on the DEC and TEC learning with the optogenetic CS. These results indicate that the cerebellum has the ability to independently support both the simple DEC, and the TEC with a trace interval of 150 or 350 ms, but not the TEC with a trace interval of 500 ms. The present results are of great importance in our understanding of the mechanisms and ability of the cerebellum in associative motor learning and memory.

Mobility is the urgent requisite of post spinal cord injury (SCI) patient. Since the alternative and compensatory approach is considered as the major function of mobility assistive device for post-SCI patients, the device should possess capability to acclimate to the ‘abnormal’ gait generate by the patients who usually undergo alternative and compensatory rehabilitation in their neural circuit. The functional ability of individual should be taking into account. Yet according to the requirement of neuro-protective treatment in post-SCI rehabilitation processing, locomotor-like activity is still an essential factor to patient. This study presents a novel concept and prototype of assistive technology base on foot control strategy to take an equilibrium between mobility and gait realization. To demonstrate that foot-induced over-ground locomotor assistive method is capable of achieving locomotor-like activity (dragging step), simulation analysis and prototype preliminary experiment have been conducted. Simulation analysis show that foot-induced assistance can allow more volitional activity compare to the hip-knee-induced assistive device. Yet the input and disturbance act on such kind of device may be increased. Surface electromyography (sEMG) from muscles of lower limb (right rectus femoris, right biceps femoris and right gluteus maximus) have been recorded during the preliminary experiment, and the mean of integrated EMG (iEMG) was used as evaluation of muscle activity. The result of the testing show that the mean of iEMG in the right gluteus maximus was reduced in the swing phase when the subject moving ahead with the prototype, but no significant change in the right rectus femoris. It may imply that foot-induced over-ground locomotor assistive device can reduce the muscle activity when patient complete locomotor-like movement and retain some amount of residual recruitment of lower limb, instead of substituting arbitrarily.

The aim of this paper was to develop a one-step in situ method to synthesize the TiN reinforced Al metallic matrix composite coatings on Ti6Al4V alloy. In this method, the Al powder and nitrogen gas were simultaneously fed into feeding nozzle during a laser nitriding process. The microstructure, microhardness and sliding wear resistance of TiN/Al coatings synthesized at different laser powers in laser nitriding were investigated. Results showed that the crack- and pore-free coatings can be made through the proposed method. However, the morphologies and distribution of TiN dendrites and wear resistance of coatings were strongly dependent on laser power used in nitriding. With increasing the laser power, the amount and density of massive TiN dendritic structure in the coating decreased and the elongated and narrow dendrites increased, leading to the increment of wear resistance of coating. When the laser power is high, the convectional flow pattern of the melt pool can be seen near the bottom of pool.

Establishment and transfer of classical eyeblink conditioning using electrical microstimulation of the hippocampus as the conditioned stimulus

Phencyclidine (PCP) is a potent drug of abuse that induces sustained schizophrenia-like symptoms in humans by blocking neurotransmission at N-methyl-d-aspartate (NMDA)-type glutamate receptors. Alterations in NMDA receptor function have been linked to numerous behavioral deficits and cognitive dysfunction. Classical eye-blink conditioning (EBC), including delay (dEBC) and trace (tEBC) paradigms, provides an effective means to study the neurobiology of associative motor learning in rodents, mammals and primates. To assess whether administration of low-dosage PCP for extended periods has prolonged effect to alter associative motor learning, in this study 19 adult cynomolgus monkeys were administered PCP (0.3mg/kg, intramuscularly) or saline twice a day for 14days. Twelve-fifteen months after PCP or saline injection, monkeys received dEBC, tEBC, or pseudo-paired training for 6 or 12 successive daily sessions, respectively. The results of this study show that percentage of conditioned response (CR) in dEBC increased as a function of training sessions in both PCP-treated and control monkeys and there was no significant CR% difference between the two groups. However, the CR timing in dEBC of PCP-treated monkeys was significantly impaired, as manifested by shorter CR peak latencies than those of the control group. PCP-treated animals showed significantly lower percentage of CR in tEBC compared to controls. PCP-treated animals were also more sensitive to outside stimuli in tEBC because the UR peak latency of PCP-treated group was significantly lower than the control group. These results indicated that cynomolgus monkeys manifested prolonged deficits in associative motor learning after long-term administration of phencyclidine.

The present experiment was designed to determine whether classical eyeblink conditioning (EBC) can be established by using electrical microstimulation of the hippocampus as a conditioned stimulus (CS) paired with an air-puff unconditioned stimulus (US). We intended to examine whether EBC transfer could occur when a CS was shifted between microstimulation of the hippocampus as a CS (Hip-CS) and tone as a CS (tone-CS) and to compare the difference in transfer effectiveness between delay EBC (dEBC) and trace EBC (tEBC). Eight groups of guinea pigs, including 4 experimental groups and 4 control groups, were included in the study. First, the experimental groups received either a Hip-CS or a tone-CS paired with a US; then, these groups were exposed to a shifted CS (tone-CS or Hip-CS) paired with the US. The control groups received the corresponding Hip-CS or tone-CS, which was, however, pseudo-paired with the US. The control groups were then shifted to the tone-CS (or Hip-CS) paired with the US. The results show that EBC can be successfully established when using microstimulation of the hippocampus as a CS paired with an air-puff US, and that the acquisition rates of EBC are higher in the experimental groups than in the control groups after switching from the Hip-CS to the tone-CS or vice versa, indicating the occurrence of learning transfer between EBC established with the Hip-CS and tone-CS. The present study also demonstrated that the EBC re-acquisition rates were remarkably higher in dEBC than in tEBC with both types of transfer, which suggests that the saving effect was more evident in dEBC than tEBC. These results significantly expand our knowledge of EBC transfer as well as the functional neural circuit underlying EBC transfer.

Learning with a stimulus from one sensory modality can facilitate subsequent learning with a new stimulus from a different sensory modality. To date, the characteristics and mechanism of this phenomenon named transfer effect still remain ambiguous. Our previous work showed that electrical stimulation of medial prefrontal cortex (mPFC) as a conditioned stimulus (CS) could successfully establish classical eyeblink conditioning (EBC). The present study aimed to (1) observe whether transfer of EBC learning would occur when CSs shift between central (mPFC electrical stimulation as a CS, mPFC-CS) and peripheral (tone as a CS, tone CS); (2) compare the difference in transfer effect between the two paradigms, delay EBC (DEBC) and trace EBC (TEBC). A total of 8 groups of guinea pigs were tested in the study, including 4 experimental groups and 4 control groups. Firstly, the experimental groups accepted central (or peripheral) CS paired with corneal airpuff unconditioned stimulus (US); then, CS shifted to the peripheral (or central) and paired with US. The control groups accepted corresponding central (or peripheral) CS and pseudo-paired with US, and then shifted CS from central (or peripheral) to peripheral (or central) and paired with US. The results showed that the acquisition rates of EBC were higher in experimental groups than in control groups after CS switching from central to peripheral or vice versa, and the CR acquisition rate was remarkably higher in DEBC than in TEBC in both transfer ways. The results indicate that EBC transfer can occur between learning established with mPFC-CS and tone CS. Memory of CS–US association for delay paradigm was less disturbed by the sudden switch of CS than for trace paradigm. This study provides new insight into neural mechanisms underlying conditioned reflex as well as the role of mPFC.

Effects of conventional exercise training of robot to stroke patients are not too satisfying, and efficient methods of training are unclear. To test how the non-rhythmical load stimulation affects cerebral cortex by analyzing the coherence between electroencephalographic signals (EEGs) and electromyographic signals (EMGs). Ten healthy subjects, all subjects have no history of neurological diseases (6 men and 4 women, mean age: 24.5 years, range: 22-28). Subjects lay on the experimental platform 75°with respect to the ground, feet on support plates and close to the ground. When non-rhythmical stimulation was performed randomly, one hinge was released and the respected braced force between the foot and support plate disappeared, which caused the corresponding ankle to extend suddenly without relative displacement between the foot and the support plate. Surface EMG signals from tibialis anterior (TA) muscles and EEG signals from cerebral cortex area Cz were recorded, and coherence between them were analyzed. The mean maximum EEG-EMG coherence of the non-rhythmical stimulation side of the ten subjects was consistent across all (9 of 10) within β range (13-30 Hz), and the average value of all in the stimulated side was 23.581Hz. While the mean maximum EEG-EMG coherence of the still side were consistent across all (9 of 10) within α range (8-13 Hz). Our findings suggest that non-rhythmical stimulation to lower limb can stimulate effectively the corresponding area of the cerebral cortex, and this idea could be applied in rehabilitation of central nervous system diseases like stroke.

The role of awareness in differential delay eyeblink conditioning (DEC) remains controversial. Here, we investigated the involvement of awareness in differential DEC with a soft or a loud tone as the conditioned stimulus (CS). In the experiment, 36 participants were trained in differential DEC with a soft tone (60 dB) or a loud tone (85 dB) as the CS, paired with a corneal air-puff as the unconditioned stimulus (US). After conditioning, awareness of the relationship between the CS and the US was assessed with a 17-item true/false questionnaire. Interestingly, during differential DEC with a soft-tone CS, a higher proportion of differential conditioned responses (CRs) was evident in participants who were aware than those who were unaware. In contrast, when a loud tone was used as the CS, the proportion of differential CRs of the aware participants did not differ significantly from those who were unaware over any of the blocks of 20 trials. In unaware participants, the percentage of differential CRs with a loud-tone CS was significantly higher than that with a soft-tone CS; however in participants classified as aware, the percentage of differential CRs with a loud-tone CS did not differ significantly from that with a soft-tone CS. The present findings suggest that awareness is critical for differential DEC when the delay task is rendered more difficult.

Research concerning impairment of associative learning during aging remains limited. The senescence-accelerated mice (SAM) prone/8 (P8) has been proposed as a useful model for the study of aging, and SAM resistant/1(SAMR1) is its control as a normal aging strain. Classical eyeblink conditioning has long been served as a model of associative learning. In order to explore the effects of aging on associative learning in SAM, the present study successively tested three paradigms of eyeblink conditioning in SAMP8 and SAMR1: classical single cue trace eyeblink conditioning (TEC), discriminative trace eyeblink conditioning and reversal learning of TEC. Behavioral performance indicated that SAMP8 could acquire limited single-cue trace eyeblink conditioning task and two-tone discrimination trace eyeblink conditioning with a relative lower acquisition rate compared to SAMR1. Both SAMP8 and SAMR1 failed to acquire reversal learning of discriminative TEC, and SAMP8' startle reflex to tone CS was lower than SAMR1. These results indicated that the impairments of aging on associative learning were incomplete in SAMP8.

The cerebellum plays an essential role in motor learning. Recently, orexins, the newfound lateral hypothalamic neuropeptides, have been found to excite Purkinje cells in the cerebellar cortex and neurons in the deep cerebellar nuclei (DCN). However, little is known about their roles in cerebellum-dependent motor learning. Therefore, the present study was designed to investigate the functional significance of hypothalamic orexinergic system during trace eyeblink conditioning, a tractable behavioral model system of cerebellum-dependent motor learning. It was revealed that the orexin 1 receptors (OXR1) were specifically localized on the soma of Purkinje cells and large DCN neurons. Furthermore, interfering with the endogenous orexins' effects on the cerebellum via the selective OXR1 antagonist SB-334867 disrupted the timing rather than the acquisition of trace conditioned eyeblink responses. In addition to the behavioral effects, the SB-334867 prevented the increase in peak amplitude of cerebellar theta oscillations with learning. These results suggest that the endogenous orexins may modulate motor learning via the activation of cerebellar OXR1.

In many previous studies, neuronal oscillations in humans under 1530 Hz activity have been observed, correlated with rhythmical motor output. In this present study, we observed the influence of non-rhythmical paroxysmal stimulation on cortical neurons by analyzing the coherence between electroencephalogram (EEG) and electromyogram (EMG). The non-rhythmical stimulation was performed by using this method that the braced force of the subjects one foot was released suddenly when the subject was standing still. The surface EMG signals from the tibialis anterior and the gastrocnemius muscles of 10 healthy subjects were recorded, and the cortical activities of contralateral cortical sensorimotor areas were also recorded simultaneously by EEG during the experiment. The data indicates that the range (10-25Hz) synchronization represents a state of the cortico-muscular network when the balance is broken which the braced force of subjects one foot is released suddenly.

Associative learning is thought to require coordinated activities among distributed brain regions. For example, to direct behavior appropriately, the medial prefrontal cortex (mPFC) must encode and maintain sensory information and then interact with the cerebellum during trace eyeblink conditioning (TEBC), a commonly-used associative learning model. However, the mechanisms by which these two distant areas interact remain elusive. By simultaneously recording local field potential (LFP) signals from the mPFC and the cerebellum in guinea pigs undergoing TEBC, we found that theta-frequency (5.0–12.0 Hz) oscillations in the mPFC and the cerebellum became strongly synchronized following presentation of auditory conditioned stimulus. Intriguingly, the conditioned eyeblink response (CR) with adaptive timing occurred preferentially in the trials where mPFC-cerebellum theta coherence was stronger. Moreover, both the mPFC-cerebellum theta coherence and the adaptive CR performance were impaired after the disruption of endogenous orexins in the cerebellum. Finally, association of the mPFC -cerebellum theta coherence with adaptive CR performance was time-limited occurring in the early stage of associative learning. These findings suggest that the mPFC and the cerebellum may act together to contribute to the adaptive performance of associative learning behavior by means of theta synchronization.

Carotid artery stenosis appears to be an independent risk factor for cognitive impairment. But the mechanisms of cognitive impairments occurring in patients with asymptomatic carotid stenosis are poorly understood. One important reason is the absence of a good carotid stenosis model. In the present study, after establishing different degrees of carotid stenosis models in aged rats successfully, we investigated their cognitive function. Wistar rat models with carotid artery stenosis were established by ligating the common carotid artery along with various gauges of needle and then removing the needle. The cognitive performance was assessed using Morris water maze (MWM) task. Cholinergic markers like acetylcholine (ACh) were evaluated in hippocampus. Neuronal damage was also studied through morphological changes. Severe carotid stenosis led to impairment in learning ability and memory retrieval, accompanied with a decrease in ACh level and morphological damage in hippocampus. Moderate carotid stenosis rats had a partial cognitive impairment, but there were no changes in ACh level and morphological damage in hippocampus. Meanwhile, mild carotid stenosis rats had no cognitive impairment compared to sham-operated rats. The results of this study suggest that different degrees of carotid stenosis induce different cognitive changes in aged rats.

It has been proposed that the medial prefrontal cortex (mPFC) is not necessary for delay eyeblink conditioning (DEC). Here, we investigated the involvement of the mPFC in DEC with a soft or loud tone as the conditioned stimulus (CS) by using electrolytic lesions or muscimol inactivation of guinea pig mPFC. Interestingly, when a soft tone was used as a CS, electrolytic lesions of the mPFC significantly retarded acquisition of the conditioned response (CR), and muscimol infusions into mPFC distinctly inhibited the acquisition and expression of CR, but had no significant effect on consolidation of well-learned CR. In contrast, both electrolytic lesions and muscimol inactivation of mPFC produced no significant deficits in the CR when a loud tone was used as the CS, or in the unconditioned response (UR) when a soft or loud tone was used as the CS. These results demonstrate that the mPFC is essential for the DEC with the soft tone CS but not for the DEC with the loud tone CS.

Learning-induced changes of synaptic ultrastructure have long been proposed as a mechanism that may contribute to support memory formation. Although recent studies have demonstrated that the interpositus nuclei (IN) play critical role in acquisition and retention of trace conditioned eyeblink responses (CRs), there is now limited evidence associating trace eyeblink conditioning with changes of synaptic ultrastructure in the IN. Here, we investigated this issue using a transmission electron microscope. Adult guinea pigs were randomly allocated to either a trace-paired, delay-paired, unpaired or exposure-only condition. The IN tissue was taken for morphological analysis 1 h after the completion of the tenth training session. Serial section analysis of synaptic ultrastructure revealed that trace eyeblink conditioning induced increases in the thickness of excitatory PSD. Classification of the synapses into shape subtypes indicated that the increased thickness of excitatory PSD was mainly attributable to increase in the concave- and convex-shaped synapses. On the contrary, trace eyeblink conditioning resulted in decreases in the thickness of inhibitory PSD. Specifically, these significant changes of PSD thickness were limited to occur in the animals with good behavioral performance. Further analysis of correlations between the trace CR performance and synaptic ultrastructural modifications showed that the thickness of excitatory PSD within the IN correlated with the peak amplitude of trace CRs, whereas the thickness of inhibitory PSD correlated with the onset latency. The present findings suggest that trace eyeblink conditioning induces structural plasticity in the IN, which may play a crucial role in acquiring and executing adaptive eyeblink movements.

This study sought to determine whether the dynamic feature of hippocampal neurons is related to the degree of complexity in complex auditory cognition. We examined the firing activity and complexity of hippocampal pyramidal cells on conscious guinea pigs during passive and active auditory discrimination tasks. The firing rate and the complexity in each sampled interspike interval (ISI) series were also computed. A larger value of complexity indicated a higher difficulty of the task or a less regularity of the discharge series. In spontaneous state, the complexity of the pyramidal cells was lower than that in low tone and low-high tone state. The complexity was increased in active discrimination state compared with passive state. The present results demonstrate that the difficulty in the auditory discriminatory task is marked by a progressive increase of complexity degree in the spike trains of pyramidal cells and cannot completely be described via mean firing rate. Moreover, the response of pyramidal cells to auditory information in different cognition states may be related to the nonlinear dynamics features, which can be quantitatively characterized by the degree of complexity. Taken together, we suggest that the degree of complexity provides an excellent experimental tool for the analysis of spikes in neural trains.

BACKGROUND/OBJECTIVES: Compelling evidence indicates a significant role for a population of CD4(+) T regulatory cells in suppressing immune responses and in maintaining immunological homeostasis. This study aims to investigate the potential role of CD4(+) CD25(HIGH) FOXP3(+) T regulatory cells in patients with chronic autoimmune urticaria and to define the characteristics of CD4(+) CD25(HIGH) FOXP3(+) cells in chronic urticaria. METHODS: We used flow cytometry to assess the expression of CD4(+) CD25(HIGH) FOXP3(+) cells in the peripheral blood mononuclear cells of patients with chronic autoimmune urticaria. RESULTS: In this study, we found that patients with chronic autoimmune urticaria have a significantly reduced frequency of CD4(+) CD25(HIGH) FOXP3(+) cells (1.39 ± 0.27% vs 2.09 ± 0.34%; P = 0.001) in their peripheral blood, accompanied by a decreased intensity of FOXP3 expression (50.13 ± 9.79 vs 68.19 ± 6.40; P < 0.001). Notably, although patients with chronic idiopathic urticaria had a reduced frequency of CD4(+) CD25(HIGH) FOXP3(+) cells (1.85 ± 0.46% vs 3.64 ± 0.48%; P < 0.001), their FOXP3 expression levels did not differ from those in healthy controls. CONCLUSIONS: Patients with chronic autoimmune urticaria displayed a reduced percentage of CD4(+) CD25(+) FOXP3(+) regulatory T cells. The results imply CD4(+) CD25(+) FOXP3(+) regulatory T cells may contribute to the autoimmune pathological process of chronic autoimmune urticaria.

The medial prefrontal cortex (mPFC) has been widely investigated for its roles in learning and memory. The present study investigated the time-limited involvement of the caudal anterior cingulate cortex (cACC) of the mPFC in the retrieval process for a simple associative motor learning, trace eyeblink conditioning (tEBC), using a 75 dB or 100 dB tone as the conditioned stimulus (CS). The GABAA receptor agonist muscimol was injected into the cACC of guinea pigs at 1 day or 4 weeks after tEBC acquisition. When muscimol was administered 1 day after tEBC acquisition, the conditioned response (CR) of the 75 dB group was severely impaired, whereas the CR of the 100 dB group exhibited no significant change relative to the control. When muscimol was administered 4 weeks after tEBC acquisition, the CR was impaired in both the 75 dB and 100 dB groups. This study indicate that the cACC of the mPFC is necessary for recent retrieval of tEBC with a low-intensity CS but not of tEBC with a high-intensity CS, whereas for remote retrieval of tEBC, the cACC of the mPFC is essential regardless of whether the CS intensity is high or low. These results support a conditional role for the mPFC in modulating recent retrieval of tEBC and a persistent role for its involvement in remote retrieval of tEBC.

The cerebellum has been proved to be essential for the acquisition of delay eyeblink conditioning, but its contribution to the acquisition of trace eyeblink conditioning (TEBC) has not been fully determined. In the present study, using chemically reversible inactivation techniques, we examined the relative contribution of ipsilateral cerebellum to the acquisition of TEBC using different time length of trace interval (TI) in guinea pigs. It was found that inactivations of the left intermediate cerebellum with a GABA(A) receptor agonist muscimol during training completely prevented the acquisition of TEBC using a relatively short (50 ms) TI, instead of the acquisition of TEBC using a relatively long (250 ms) TI. However, inactivations of the left intermediate cerebellum totally abolished the well-established left trace conditioned eyeblink responses (CRs) regardless of the time length of TI. These results suggested that while the ipsilateral cerebellum is essential for the expression of trace CRs, its contribution to the acquisition of trace CRs appears to mainly depend on the time length of TI.

The aim of this study was to evaluate the relative contributions of the ipsilateral and contralateral cerebellum to the acquisition of unilateral classical eyeblink conditioning (EBCC). The unilateral EBCC was achieved using a binaural tone conditioned stimulus (CS) paired with a left airpuff unconditioned stimulus (US). A high-resolution potentiometer was used to monitor eyeblink responses. Guinea pigs received one CS-US session followed by three CS-US sessions (sessions 2 to 4), during which microinjections of muscimol, a GABAA receptor agonist, were performed to reversibly inactivate the cerebellum unilaterally prior to training. To test whether any learning had occurred during these inactivation sessions, training was continued for six more CS-US sessions (sessions 5 to 10) without any inactivation. Animals with inactivation of the left cerebellum had no signs of left conditioned response (CR) during sessions 2 to 4, and their CR acquisition during sessions 5 to 10 was not distinguishable from that of control animals during sessions 2 to 7. In contrast, animals with inactivation of the right cerebellum acquired left CRs during sessions 2 to 4, although their CR acquisition was significantly retarded during session 2. In addition, microinjections of muscimol into the right cerebellum did not affect left neuro-behavioral activity. Finally, microinjections of muscimol into either the left or the right cerebellum did not affect the performance of tone-airpuff evoked unconditioned response (UR). In contrast to the essential role of the ipsilateral cerebellum, the contralateral cerebellum is potentially involved in the acquisition of unilateral EBCC during the early stage of training.

Objective To clarify the firing characteristics of the hippocampal pyramidal cells and interneurons in the auditory discrimination cognition. Methods Thirteen guinea pigs were studied by the paired (active cognition group, n = 10) or unpaired (passive cognition group, n = 3) training with 1 kHz (CS+) and 500 Hz tones (CS−) and the air puff (US) applied 250 ms after the CS+ onset. Results In active group, 32 pyramidal cells showed exciting response to the CS+ tone, 16 cells inhibited response and 4 cells revealed no response to the high frequency tone and 18 interneurons almost unchanged. In passive group, the pyramidal cells responded to the tone casually and 10 out of the 13 interneurons remained invariably. Conclusion The result suggests that the pyramidal cells play a major role in coding auditory information by the networks, and the interneuons may modulate it via forward and feedback.

Traumatic brain injury (TBI) often results in multiple neuropsychiatric sequelae, including cognitive, emotional, and behavioral problems. Among them, depression is a common psychiatric symptom, and links to poorer recovery. Amantadine, as an antiparkinsonian, increases dopamine release, and blocks dopamine reuptake, but has recently received attention for its effectiveness as an antidepressant. In the present study, we first induced a post-TBI depression rat model to probe the efficacy of amantadine therapy in reducing post-TBI depression. The DA concentration in the striatum of the injured rats, as well as the degeneration and apoptosis of dopaminergic neurons in the substantia nigra (SN), were checked along with the depression-like behavior. The results showed that amantadine therapy could significantly ameliorate the depression-like behavior, improving the DA level in the striatum and decreasing the degeneration and apoptosis of dopaminergic neurons in the SN. The results indicated that the anti-depression effect may result from the increase of extracellular DA concentration in the striatum and/or the indirect neuroprotection on the dopaminergic neurons in the SN. We conclude that DA plays a critical role in post-TBI depression, and that amantadine shows its potential value in anti-depression treatment for TBI.

Classical eyeblink conditioning (EBC), a simple form of associative learning, has long been served as a model for motor learning and modulation. The neural circuitry of EBC has been studied in detail in rabbits. However, its underlying molecular mechanisms remain unclear. The advent of mouse transgenics has generated new perspectives on the studies of the neural substrates and molecular mechanisms essential for EBC. Results about EBC in mice differ in some aspects from those obtained in other mammals. Here, we review the current studies about the neural circuitry and molecular mechanisms underlying delay and trace EBC in mice. We conclude that brainstem-cerebellar circuit plays an essential role in DEC while the amygdala modulates this process, and that the medial prefrontal cortex (mPFC) as a candidate is involved in the extra-cerebellar mechanism underlying delay eyeblink conditioning (DEC) in mice. We propose the Amygdala-Cerebellum-Prefrontal Cortex-Dynamic-Conditioning Model (ACPDC model) for DEC in mice. As to trace eyeblink conditioning (TEC), the forebrain regions may play an essential role in it, whereas cerebellar cortex seems to be out of the neural circuitry in mice. Moreover, the molecular mechanisms underlying DEC and TEC in mice differ from each other. This review provides some new information and perspectives for further research on EBC.

Aims Previous studies in our laboratory have demonstrated the increased neuronal apoptosis in the hippocampus and abnormal hippocampal morphology after severe stress, which directly correlates to the pathogenesis of post-traumatic stress disorder (PTSD). This study aims to investigate the effects of Lycium barbarum polysaccharide (LBP) on intrusive memory of posttraumatic stress in rats, and to analyze the mechanism of regeneration/apoptosis balance in the hippocampal neurons. Main methods The experimental rats received 20 inescapable electric foot shocks in an enclosed box for six times in three days. The rats were treated by intragastric administration of LBP (20 mg/kg/day) for 3 days before stress in the stress plus prophylactic group, and for 28 days after stress in the stress plus therapeutic group. The emotion, intrusive memory-related behavior (freezing, open field, pain latency, spatial cognition), hippocampus cell morphology, and relation of neurogenesis and apoptosis in dental gyrus of the hippocampus were observed. The hippocampus volume was evaluated by stereology. Meanwhile, the neurogenesis and apoptosis were analyzed with 5-bromo-2′-deoxyuridine and terminal deoxylnucleotidyl transferase mediated-dUTP nick end labeling (TUNEL) method. Key findings The treatment of LBP in pre-stress and post-stress had obvious beneficial effect on the behaviors and neurogenesis. The stressed rats showed improvement of intrusive memory related cognition defect, alleviation of the apoptosis in the hippocampus and recovery for the neurogenesis, which was related to the hippocampus volume after LBP treatment. Significance LBP treatment might effectively improve the traumatic cognition defect induced by severe stress and be useful for the intrusive memory-related cognition recovery.

Stimulus-evoked theta oscillations are observed in the medial prefrontal cortex (mPFC) when executing a variety of learning tasks. Here, we aimed to further determine whether spontaneous theta-band (5.0-10.0 Hz) oscillations in the mPFC predicted the subsequent behavioral performance in trace eyeblink conditioning (TEBC), in which the conditioned stimulus (CS) was separated from the unconditioned stimulus (US) by 500 ms trace interval. By recording local field potentials (LFP) signals in the guinea pigs performing the TEBC task, we found that, a higher mPFC relative theta ratio [theta/(delta+beta)] during the baseline (850-ms period prior to the onset of the CS) was predictive of higher magnitude and more adaptive timing rather than faster acquisition of trace conditioned eyeblink responses (CR). However, the prediction of baseline mPFC theta activity was time-limited to the well-learning stage. Additionally, the relative power of mPFC theta activity did not correlate with the CR performance if the trace interval between the CS and the US was shortened to 100 ms. These results suggest that the brain state in which the baseline mPFC theta activity is present or absent is detrimental for the subsequent performance of trace CRs especially when the asymptotic learning state is achieved.

Behavioral studies have demonstrated that both medial prefrontal cortex (mPFC) and cerebellum play critical roles in trace eyeblink conditioning. However, little is known regarding the mechanism by which the two brain regions interact. By use of electrical stimulation of the caudal mPFC as a conditioned stimulus, we show evidence that persistent outputs from the mPFC to cerebellum are necessary and sufficient for the acquisition and expression of a trace conditioned response (CR)-like response. Specifically, the persistent outputs of caudal mPFC are relayed to the cerebellum via the rostral part of lateral pontine nuclei. Moreover, interfering with persistent activity by blockade of the muscarinic Ach receptor in the caudal mPFC impairs the expression of learned trace CRs. These results suggest an important way for the caudal mPFC to interact with the cerebellum during associative motor learning.

The role of the hippocampus in delay eyeblink conditioning (DEC) remains controversial. Here, we investigated the involvement of the hippocampus in DEC with a soft tone as the conditioned stimulus (CS) by using electrolytic lesions or muscimol inactivation of guinea pig dorsal hippocampus. Interestingly, when a soft tone was used as a CS, electrolytic lesions of the hippocampus significantly retarded acquisition of the conditioned response (CR), and muscimol infusions into hippocampus distinctly inhibited the acquisition and expression of CR, but had no significant effect on consolidation of well-learned CR. In contrast, both electrolytic lesions and muscimol inactivation of hippocampus produced no significant deficits in the CR when a loud tone was used as the CS. These results demonstrate that the hippocampus is essential for the DEC when the delay task was rendered more difficult.

To determine whether electrical stimulation of caudal medial prefrontal cortex (mPFC) as conditioned stimulus (CS) paired with airpuff unconditioned stimulus (US) was sufficient for establishing eyeblink conditioning in guinea pigs, and whether it was dependent on cerebellar interpositus nucleus.Thirty adult guinea pigs were divided into 3 conditioned groups, and trained on the delay eyeblink conditioning, short-trace eyeblink conditioning, and long-trace eyeblink conditioning paradigms, respectively, in which electrical stimulation of the right caudal mPFC was used as CS and paired with corneal airpuff US. A pseudo conditioned group of another 10 adult guinea pigs was given unpaired caudal mPFC electrical stimulation and the US. Muscimol (1 μg in 1 μL saline) and saline (1 μL) were infused into the cerebellar interpositus nucleus of the animals through the infusion cannula on d 11 and 12, respectively.The 3 eyeblink conditioning paradigms have been successfully established in guinea pigs. The animals acquired the delay and short-trace conditioned responses more rapidly than long-trace conditioned responses. Muscimol infusion into the cerebellar interpositus nucleus markedly impaired the expression of the 3 eyeblink conditioned responses.Electrical stimulation of caudal mPFC is effective CS for establishing eyeblink conditioning in guinea pigs, and it is dependent on the cerebellar interpositus nucleus.

Compelling evidence indicates a significant role for a population of CD4(+) T regulatory cells in suppressing immune responses and in maintaining immunological homeostasis. This study aims to investigate the potential role of CD4(+) CD25(HIGH) FOXP3(+) T regulatory cells in patients with chronic autoimmune urticaria and to define the characteristics of CD4(+) CD25(HIGH) FOXP3(+) cells in chronic urticaria.We used flow cytometry to assess the expression of CD4(+) CD25(HIGH) FOXP3(+) cells in the peripheral blood mononuclear cells of patients with chronic autoimmune urticaria.In this study, we found that patients with chronic autoimmune urticaria have a significantly reduced frequency of CD4(+) CD25(HIGH) FOXP3(+) cells (1.39 ± 0.27% vs 2.09 ± 0.34%; P = 0.001) in their peripheral blood, accompanied by a decreased intensity of FOXP3 expression (50.13 ± 9.79 vs 68.19 ± 6.40; P0.001). Notably, although patients with chronic idiopathic urticaria had a reduced frequency of CD4(+) CD25(HIGH) FOXP3(+) cells (1.85 ± 0.46% vs 3.64 ± 0.48%; P0.001), their FOXP3 expression levels did not differ from those in healthy controls.Patients with chronic autoimmune urticaria displayed a reduced percentage of CD4(+) CD25(+) FOXP3(+) regulatory T cells. The results imply CD4(+) CD25(+) FOXP3(+) regulatory T cells may contribute to the autoimmune pathological process of chronic autoimmune urticaria.

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To develop simple but reliable intracellular labelling method for high-resolution visualization of the fine structure of single neurons in brain slice with thickness of 500 microm.Biocytin was introduced into neurons in 500 microm-thickness brain slices while blind whole cell recording. Following processed for histochemistry using the avidin-biotin-complex method, stained slices were mounted in glycerol on special glass slides. Labelled cells were digital photomicrographed every 30 microm and reconstructed with Adobe Photoshop software.After histochemistry, limited background staining was produced. The resolution was so high that fine structure, including branching, termination of individual axons and even spines of neurons could be identified in exquisite detail with optic microscope. With the help of software, the neurons of interest could be reconstructed from a stack of photomicrographs.The modified method provides an easy and reliable approach to revealing the detailed morphological properties of single neurons in 500 microm-thickness brain slice. Without requisition of special equipment, it is suited to be broadly applied.