PPy/SWCNTs-Modified Microelectrode Array for Learning and Memory Model Construction through Electrical Stimulation and Detection of In Vitro Hippocampal Neuronal Network.

The learning and memory functions of the brain remain unclear, which are in urgent need for the detection of both a single cell signal with high spatiotemporal resolution and network activities with high throughput. Here, an in vitro microelectrode array (MEA) was fabricated and further modified with polypyrrole/carboxylated single-walled carbon nanotubes (PPy/SWCNTs) nanocomposites as the interface between biological and electronic systems. The deposition of the nanocomposites significantly improved the performance of microelectrodes including low impedance (60.3 ± 28.8 k Ω), small phase delay (-32.8 ± 4.4°), and good biocompatibility. Then the modified MEA was used to apply learning training and test on hippocampal neuronal network cultured for 21 days through electrical stimulation, and multichannel electrophysiological signals were recorded simultaneously. During the process of learning training, the stimulus/response ratio of the hippocampal learning population gradually increased and the response time gradually decreased. After training, the mean spikes in burst, number of bursts, and mean burst duration increased by 53%, 191%, and 52%, respectively, and the correlation of neurons in the network was significantly enhanced from 0.45 ± 0.002 to 0.78 ± 0.002. In addition, the neuronal network basically retained these characteristics for at least 5 h. These results indicated that we have successfully constructed a learning and memory model of hippocampal neurons on the in vitro MEA, contributing to understanding learning and memory based on synaptic plasticity. The proposed PPy/SWCNTs-modified in vitro MEA will provide a promising platform for the exploration of learning and memory mechanism and their applications in vitro.

Ontogeny of the digestive tract and enzymes in rock bream Oplegnathus fasciatus (Temminck et Schlegel 1844) larvae.

Histological development of the digestive tract and specific activities of three digestive enzymes (trypsin, alkaline phosphatase, and pepsin) were studied in rock bream Oplegnathus fasciatus from hatching to 50 days after hatching (DAH). At hatching, the digestive tract appeared as an undifferentiated straight tube and differentiated into the buccopharynx, esophagus, stomach, intestine, and rectum at mouth opening by 3 DAH. The taste bud and mandibular teeth were present in the buccopharyx at 8 DAH. The goblet cells appeared in the esophagus at 8 DAH and in the buccopharyx at 9 DAH. The stomach anlage was formed at 2 DAH and developed into cardia, fundus, and pylorus at 14 DAH. The gastric glands were visible at 16 DAH, and the pepsin was firstly detected on 22 DAH. At 2 DAH, the intestinal valve appeared and divided the intestine into anterior intestine (AI) and posterior intestine (PI). The rectum was differentiated from the PI at 3 DAH. The supranuclear vacuoles were visible in the rectum by 6 DAH, and the lipid inclusions were present in the AI at 8 DAH. The alkaline phosphatase was detected at 1 DAH, and the increase in its activity indicated the maturation of the intestine after 40 DAH. The hepatocytes and pancreatic cells were differentiated from the blast cells at 2 DAH, and the acidophilic zymogen granules in the exocrine pancreas were observed simultaneously. The trypsin was detected by 1 DAH and increased to the maximum at 19 DAH, followed by a decrease as the stomach became functional.

A Neural Sensor with a Nanocomposite Interface for the Study of Spike Characteristics of Hippocampal Neurons under Learning Training.

Both the cellular- and population-level properties of involved neurons are essential for unveiling the learning and memory functions of the brain. To give equal attention to these two aspects, neural sensors based on microelectrode arrays (MEAs) have been in the limelight due to their noninvasive detection and regulation capabilities. Here, we fabricated a neural sensor using carboxylated graphene/3,4-ethylenedioxythiophene:polystyrenesulfonate (cGO/PEDOT:PSS), which is effective in sensing and monitoring neuronal electrophysiological activity in vitro for a long time. The cGO/PEDOT:PSS-modified microelectrodes exhibited a lower electrochemical impedance (7.26 ± 0.29 kΩ), higher charge storage capacity (7.53 ± 0.34 mC/cm2), and improved charge injection (3.11 ± 0.25 mC/cm2). In addition, their performance was maintained after 2 to 4 weeks of long-term cell culture and 50,000 stimulation pulses. During neural network training, the sensors were able to induce learning function in hippocampal neurons through precise electrical stimulation and simultaneously detect changes in neural activity at multiple levels. At the cellular level, not only were three kinds of transient responses to electrical stimulation sensed, but electrical stimulation was also found to affect inhibitory neurons more than excitatory neurons. As for the population level, changes in connectivity and firing synchrony were identified. The cGO/PEDOT:PSS-based neural sensor offers an excellent tool in brain function development and neurological disease treatment.

High-Throughput PEDOT:PSS/PtNPs-Modified Microelectrode Array for Simultaneous Recording and Stimulation of Hippocampal Neuronal Networks in Gradual Learning Process.

When it comes to mechanisms of brain functions such as learning and memory mediated by neural networks, existing multichannel electrophysiological detection and regulation technology at the cellular level does not suffice. To address this challenge, a 128-channel microelectrode array (MEA) was fabricated for electrical stimulation (ES) training and electrophysiological recording of the hippocampal neurons in vitro. The PEDOT:PSS/PtNPs-coated microelectrodes dramatically promote the recording and electrical stimulation performance. The MEA exhibited low impedance (10.94 ± 0.49 kohm), small phase delay (-12.54 ± 0.51°), high charge storage capacity (14.84 ± 2.72 mC/cm2), and high maximum safe injection charge density (4.37 ± 0.22 mC/cm2), meeting the specific requirements for training neural networks in vitro. A series of ESs at various frequencies was applied to the neuronal cultures in vitro, seeking the optimum training mode that enables the neuron to display the most obvious plasticity, and 1 Hz ES was determined. The network learning process, including three consecutive trainings, affected the original random spontaneous activity. Along with that, the firing pattern gradually changed to burst and the correlation and synchrony of the neuronal activity in the network have progressively improved, increasing by 314% and 240%, respectively. The neurons remembered these changes for at least 4 h. Collectively, ES activates the learning and memory functions of neurons, which is manifested in transformations in the discharge pattern and the improvement of network correlation and synchrony. This study offers a high-performance MEA revealing the underlying learning and memory functions of the brain and therefore serves as a useful tool for the development of brain functions in the future.

Highly Flexible, Stretchable, and Self-Powered Strain-Temperature Dual Sensor Based on Free-Standing PEDOT:PSS/Carbon Nanocoils-Poly(vinyl) Alcohol Films.

The wearable and self-powered sensors with multiple functions are urgently needed for energy saving devices, economical convenience, and artificial human skins. It is a meaningful idea to convert excess heat sources into power supplies for wearable sensors. In this report, we have fabricated a series of free-standing self-powered temperature-strain dual sensors based on poly(3,4-ethylenedioxythiophene) polystyrenesulfonate (PEDOT:PSS)/carbon nanocoils (CNCs)-poly(vinyl) alcohol composite films by a simple drop casting method. The Seebeck coefficients of the composite films were measured to be 19 µV/K. The sensor, with the addition of CNCs, showed a superior sensing performance to that without CNCs. PEDOT:PSS is used to provide a thermoelectric power to detect temperature changes and strain deformations. The minimum detect limit for the temperature difference was 0.3 K. Under a constant temperature gradient of 30 K, strains from 1 to 10% were detected without any external power supply. The films can be easily made into an array to detect the temperature of the fingers and motions of the wrist by attaching it to the human wrist directly. For the first time, due to the independent action of the thermoelectric material and strain sensing material, the thermoelectric voltage which is generated by a constant temperature difference is maintained under different strains. This kind of free-standing self-powered multifunctional sensors has great application prospects in the fields of healthcare and artificial intelligence in the future.

SWCNTs/PEDOT:PSS-Modified Microelectrode Arrays for Dual-Mode Detection of Electrophysiological Signals and Dopamine Concentration in the Striatum under Isoflurane Anesthesia.

Accurate detection of the degree of isoflurane anesthesia during a surgery is important to avoid the risk of overdose isoflurane anesthesia timely. To address this challenge, a four-shank implantable microelectrode array (MEA) was fabricated for the synchronous real-time detection of dual-mode signals [electrophysiological signal and dopamine (DA) concentration] in rat striatum. The SWCNTs/PEDOT:PSS nanocomposites were modified onto the MEAs, which significantly improved the electrical and electrochemical performances of the MEAs. The electrical performance of the modified MEAs with a low impedance (16.20 ± 1.68 kΩ) and a small phase delay (-27.76 ± 0.82°) enabled the MEAs to detect spike firing with a high signal-to-noise ratio (> 3). The electrochemical performance of the modified MEAs with a low oxidation potential (160 mV), a low detection limit (10 nM), high sensitivity (217 pA/µM), and a wide linear range (10 nM-72 µM) met the specific requirements for DA detection in vivo. The anesthetic effect of isoflurane was mediated by inhibiting the spike firing of D2_SPNs (spiny projection neurons expressing the D2-type DA receptor) and the broadband oscillation rhythm of the local field potential (LFP). Therefore, the spike firing rate of D2_SPNs and the power of LFP could reflect the degree of isoflurane anesthesia together. During the isoflurane anesthesia-induced death procedure, we found that electrophysiological activities and DA release were strongly inhibited, and changes in the DA concentration provided more details regarding this procedure. The dual-mode recording MEA provided a detection method for the degree of isoflurane anesthesia and a prediction method for fatal overdose isoflurane anesthesia.