Anatomical characterization of vagal nodose afferent innervation and ending morphologies at the murine heart using a transgenic approach.

Heart is an extensively innervated organ and its function is strictly coordinated by autonomic neural circuits. After pathological events such as myocardial infarction (MI), cardiac nerves undergo a structural and functional remodeling contributing to cardiac dysfunction. Although the efferent component of the cardiac nerves has been well described, sensory innervation of the heart has not been defined in detail. Considering its importance, comprehensive description of vagal afferent innervation on the whole heart would enable a better description of autonomic imbalances manifesting as sympathoexcitation and vagal withdrawal in post-ischemic states. To address this issue, we globally mapped the vagal nodose afferent fibers innervating the whole murine heart with unprecedented resolution. By using the Phox2b-Cre::tdTomato transgenic mouse line, we described the detailed distribution and distinct vagal sensory ending morphologies at both the dorsal and ventral sides of the mouse heart. By neural tracing analysis, we quantitated the distribution and prevalence of vagal afferent nerve fibers with varying diameters across dorsal and ventral surfaces of the heart. Moreover, we demonstrated that vagal afferents formed flower spray and end-net-like endings within the atria and ventricles. As distinct from the atria, vagal afferents formed intramuscular array-like endings within the ventricles. Furthermore, we showed that vagal afferents undergo structural remodeling by forming axonal sprouts around the infarct area in post-MI hearts. These findings improve our understanding of the potential effect of vagal afferent remodeling on autonomic imbalance and generation of cardiac arrhythmias and could prospectively contribute to the development of more effective neuromodulatory therapies.

Defining optimal enzyme and matrix combination for replating of human induced pluripotent stem cell-derived cardiomyocytes at different levels of maturity.

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) create an unlimited cell source for basic and translational research. Depending on the maturity of cardiac cultures and the intended applications, obtaining hiPSC-CMs as a single-cell, monolayer or three-dimensional clusters can be challenging. Here, we defined strategies to replate hiPSC-CMs on early days (D15-30) or later more mature (D60-150) differentiation cultures. After generation of hiPSCs and derivation of cardiomyocytes, four dissociation reagents Collagenase A/B, Collagenase II, TrypLE, EDTA and five different extracellular matrix materials Laminin, iMatrix-511, Fibronectin, Matrigel, and Geltrex were comparatively evaluated by imaging, cell viability, and contraction analysis. For early cardiac differentiation cultures mimicking mostly the embryonic stage, the highest adhesion, cell viability, and beating frequencies were achieved by treatment with the TrypLE enzyme. Video-based contraction analysis demonstrated higher beating rates after replating compared to before treatment. For later differentiation days of more mature cardiac cultures, dissociation with EDTA and replating cells on Geltrex or Laminin-derivatives yielded better recovery. Cardiac clusters at various sizes were detected in several groups treated with collagenases. Collectively, our findings revealed the selection criteria of the dissociation approach and coating matrix for replating iPSC-CMs based on the maturity and the requirements of further downstream applications.

Event-related EEG oscillatory responses elicited by dynamic facial expression.

BACKGROUND: Recognition of facial expressions (FEs) plays a crucial role in social interactions. Most studies on FE recognition use static (image) stimuli, even though real-life FEs are dynamic. FE processing is complex and multifaceted, and its neural correlates remain unclear. Transitioning from static to dynamic FE stimuli might help disentangle the neural oscillatory mechanisms underlying face processing and recognition of emotion expression. To our knowledge, we here present the first time-frequency exploration of oscillatory brain mechanisms underlying the processing of dynamic FEs. RESULTS: Videos of joyful, fearful, and neutral dynamic facial expressions were presented to 18 included healthy young adults. We analyzed event-related activity in electroencephalography (EEG) data, focusing on the delta, theta, and alpha-band oscillations. Since the videos involved a transition from neutral to emotional expressions (onset around 500 ms), we identified time windows that might correspond to face perception initially (time window 1; first TW), and emotion expression recognition subsequently (around 1000 ms; second TW). First TW showed increased power and phase-locking values for all frequency bands. In the first TW, power and phase-locking values were higher in the delta and theta bands for emotional FEs as compared to neutral FEs, thus potentially serving as a marker for emotion recognition in dynamic face processing. CONCLUSIONS: Our time-frequency exploration revealed consistent oscillatory responses to complex, dynamic, ecologically meaningful FE stimuli. We conclude that while dynamic FE processing involves complex network dynamics, dynamic FEs were successfully used to reveal temporally separate oscillation responses related to face processing and subsequently emotion expression recognition.