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Exocytosis is an essential cellular trafficking process integral to the proper distribution and function of a plethora of molecules, including transporters, receptors, and enzymes. Moreover, incorrect protein targeting can lead to pathological conditions. Recently, the application of evanescent wave microscopy has allowed us to image the final steps of exocytosis. However, spatio-temporal analysis of fusion of constitutive vesicular traffic with the plasma membrane has not been systematically performed. Also, the spatial sites and times of vesicle fusion have not yet been analyzed together. In addition, more formal tests are required in testing biological hypotheses, rather than visual inspection combined with statistical descriptives. Ripley K-functions are used to examine the joint and marginal behavior of locations and fusion times. Semiautomatic detection and mapping of constitutive fusion sites reveals spatial and temporal clustering, but no dependency between the locations and times of fusion events. Our novel approach could be translated to other studies of membrane trafficking in health and diseases such as diabetes.

The lateral nucleus of the amygdala (LA) is a site of convergence for auditory (conditioned stimulus) and footshock (unconditioned stimulus) inputs during fear conditioning. The auditory pathways to LA are well characterized, but less is known about the pathways through which footshock is transmitted. Anatomical tracing and physiological recording studies suggest that the posterior intralaminar thalamic nucleus, which projects to LA, receives both auditory and somatosensory inputs. In the present study we examined the expression of the immediate-early gene c-fos in the LA in rats in response to footshock stimulation. We then determined the effects of posterior intralaminar thalamic lesions on footshock-induced c-Fos expression in the LA. Footshock stimulation led to an increase in the density of c-Fos-positive cells in all LA subnuclei in comparison to controls exposed to the conditioning box but not shocked. However, some differences among the dorsolateral, ventrolateral and ventromedial subnuclei were observed. The ventrolateral subnucleus showed a homogeneous activation throughout its antero-posterior extension. In contrast, only the rostral aspect of the ventromedial subnucleus and the central aspect of the dorsolateral subnucleus showed a significant increment in c-Fos expression. The density of c-Fos-labeled cells in all LA subnuclei was also increased in animals placed in the box in comparison to untreated animals. Unilateral electrolytic lesions of the posterior intralaminar thalamic nucleus and the medial division of the medial geniculate body reduced footshock-induced c-Fos activation in the LA ipsilateral to the lesion. The number of c-Fos labeled cells on the lesioned side was reduced to the levels observed in the animals exposed only to the box. These results indicate that the LA is involved in processing information about the footshock unconditioned stimulus and receives this kind of somatosensory information from the posterior intralaminar thalamic nucleus and the medial division of the medial geniculate body. KeywordsEmotional learning; fear conditioning; pain; freezing; electrolytic lesions; somatosensory thalamus Address NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptIn recent years classical fear conditioning has become a leading model for studying the neural mechanisms of learning and memory in mammals (LeDoux, 2000;Maren, 2005). In this behavioral paradigm, a neutral (innocuous) conditioned stimulus (CS), such as a tone, is presented in association with an aversive unconditioned stimulus (US), such as a mild footshock. After a few such paired presentations, the animal begins to respond defensively to the neutral stimulus. Considerable evidence suggests the lateral nucleus of the amygdala (LA) is a site of auditory CS and somatosensory US convergence (Romanski et al., 1993;Bordi and LeDoux, 1994b) and is crucial for the formation of the association between the CS and US (Blair et al, 2001;Fanselow and LeDoux, 1999;LeDoux, 2000;Maren, 2001Maren, , 2005; but...

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Emotional response in aging is typically studied using the dimensional or the discrete models of emotion. Moreover, it is typically studied using subjective or physiological variables but not using both perspectives simultaneously. Additionally, tenderness is neglected in emotion induction procedures with older adults, with the present work being the first to include the study of physiological tenderness using film clips. This study integrated two separate approaches to emotion research, comparing 68 younger and 39 older adults and using a popular set of film clips to induce tenderness, amusement, anger, fear, sadness and disgust emotions. The direction of subjective emotional patterns was evaluated with self-reports and that of physiological emotional patterns was evaluated with a wearable emotion detection system. The findings suggest a dual-process framework between subjective and physiological responses, manifested differently in young and older adults. In terms of arousal, the older adults exhibited higher levels of subjective arousal in negative emotions and tenderness while young adults showed higher levels of physiological arousal in these emotions. These findings yield information on the multidirectionality of positive and negative emotions, corroborating that emotional changes in the adult lifespan appear to be subject to the relevance of the emotion elicitor to each age group. It is widely recognized that aging is associated with changes in the direction of emotional patterns 1 , but the knowledge of how the processes involved in emotions change as we age is still incomplete 2. In recent decades, much work has been done to establish different theories to explain emotional aging. Emotional theories such as the Socioemotional Selectivity Theory (SST 3 ; have underlined a positivity effect, whereby older adults tend to give primacy to achieving emotional gratification 1 , and avoid or mitigate exposure to negative situations 4,5. This model has been empirically studied in psychological and neuroscientific research 2,6,7 and is currently one of the most widely accepted models. However, Schweizer et al. 8 found that older adults exhibited decline in the capacity to down-regulate negative affect with film clips, which would suggest an absence of the positivity effect. Other authors have suggested that differences in the emotional experience depend on the stimuli used in the Mood Induction Procedure (MIP) 9. For example, different studies have found that older adults present greater emotional responses when the eliciting stimuli involve irrevocable personal loss or deal with social injustice 10,11. Compared with the literature on subjective emotional patterns, that on physiological patterns in aging is limited. A meta-analytical review of 31 studies using different MIPs conducted in laboratory settings found that the young and older adults' physiological patterns were similar, although the amplitude of this response was

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For the sake of establishing the neural correlates of phrase quadrature perception in harmonic rhythm, a musical experiment has been designed to induce music-evoked stimuli related to one important aspect of harmonic rhythm, namely the phrase quadrature. Brain activity is translated to action through electroencephalography (EEG) by using a brain-computer interface. The power spectral value of each EEG channel is estimated to obtain how power variance distributes as a function of frequency. The results of processing the acquired signals are in line with previous studies that use different musical parameters to induce emotions. Indeed, our experiment shows statistical differences in theta and alpha bands between the fulfillment and break of phrase quadrature, an important cue of harmonic rhythm, in two classical sonatas.