MEA-based recording of neuronal activity in vitro

Based on the advantages of MEA-based recording, developmental changes of spontaneous activity and tetanus-induced modification of evoked activity were studied. Rat cortical neurons were cultured on MEAs and the spontaneous activity was continuously monitored for two months. The activity started a few days after plating. During the second week, the cultures generated periodic synchronized bursts, which were the characteristic properties of cortical neurons in vitro. In about one month, the cultured networks reached a steady state. Between these two, we found a critical period during which only weak activities were generated. This critical period might reflect the transition from immature networks to mature networks including precisely controlled excitatory and inhibitory synapses. We could elicit clear evoked responses with high reproducibility in mature cultures. A focal tetanic stimulation was applied to the mature cultures and how the tetanus affects 64 kinds of evoked activity was studied. The evoked responses showed bi-directional changes in their propagation patterns, potentiation and depression. These induced changes reflected the correlation properties with the tetanized activity pattern. The next step will be the combination of long-term recording and multi-site stimulation. How long does the induced change last, as well as how additional strong activity affects the previously induced changes, will be studied.     

Chronic extracellular recording of fetal medullary neuronal activity

A technique for chronic extracellular recording of neuronal activity in the medullary region of fetal sheep during different sleep states is described. Of 51 recorded neurones, 54% were associated with respiration, blood pressure changes, or muscle electromyograms, and 46% were nonspecific. During apnea (nonrapid-eye-movement sleep) both inspiratory and expiratory neurons had random intermittent activity. Thus apnea is not associated with tonic expiratory neuronal activity.     

Optimal signal filtering parameters for tetrode recording of neuronal activity

Extracellular recording of neuronal spiking is the main method of investigation of involvement of neurons in behavioral tasks. Development of multichannel electrodes made it possible to simultaneously record activity of the same group of neurons from different locations in the brain tissue. That method allows the researches to distinguish spiking of simultaneously recorded neurons by individual set of projection coefficients of amplitude parameters on axes corresponding to different channels of the multichannel electrode. We tested the possibility of effective separation of single unit spiking streams from multiunit activity recorded by tetrode and subjected to different filtering. We described the main limitations for effective spike identification and determined the optimal band of signal filtering for tetrode recording.     

A compact multiphoton 3D imaging system for recording fast neuronal activity

We constructed a simple and compact imaging system designed specifically for the recording of fast neuronal activity in a 3D volume. The system uses an Yb:KYW femtosecond laser we designed for use with acousto-optic deflection. An integrated two-axis acousto-optic deflector, driven by digitally synthesized signals, can target locations in three dimensions. Data acquisition and the control of scanning are performed by a LeCroy digital oscilloscope. The total cost of construction was one order of magnitude lower than that of a typical Ti:sapphire system. The entire imaging apparatus, including the laser, fits comfortably onto a small rig for electrophysiology. Despite the low cost and simplicity, the convergence of several new technologies allowed us to achieve the following capabilities: i) full-frame acquisition at video rates suitable for patch clamping; ii) random access in under ten microseconds with dwelling ability in the nominal focal plane; iii) three-dimensional random access with the ability to perform fast volume sweeps at kilohertz rates; and iv) fluorescence lifetime imaging. We demonstrate the ability to record action potentials with high temporal resolution using intracellularly loaded potentiometric dye di-2-ANEPEQ. Our design proffers easy integration with electrophysiology and promises a more widespread adoption of functional two-photon imaging as a tool for the study of neuronal activity. The software and firmware we developed is available for download at http://neurospy.org/ under an open source license.     

Telemetric motor activity in children. A preliminary study

A lightweight telemetric mobility sensing system was used to study the relationship between high levels of motor activity during free-play and school performance. Among the 21 normal children, there was a significant correlation between high ankle motor activity during free-play, poor school achievement, the presence of neurological soft signs, and a poor self-image. Those normals whose free-play ankle activity was above the mean, also had significantly more errors and performed at a significantly lower level on the Bender Visual Motor Gestalt Test than children whose activity was below the mean. This preliminary study suggests that the telemetric mobility sensing system can be easily applied to children to assess clinically relevant components of psychomotor activity.     

Telemetric Electrocardiography

By means of a satellite transmitter nearby the subject an electrocardiogram can be obtained by telemetry in a hospital electrocardiographic (ECG) laboratory from an area of approximately 20 miles in diameter within the city of London, Ontario. The technique employed in this preliminary trial produced excellent tracings, quite comparable to tracings obtained using the same leads with direct recording. Ninety-four trials were performed on 63 subjects, 37 of whom were patients with a suspected or known myocardial abnormality. A normal group of 26 healthy subjects, 16 of whom were strenuously exercised on a bicycle ergometer to the point of physical exhaustion, were also studied. Monitoring patients during ordinary daily activities has proved extremely useful in the detection and diagnosis of cardiac arrhythmias, intermittent conduction disturbances, latent coronary artery disease, and asymptomatic heart disease.     

Compact micromanipulator for the chronic extracellular recording of neuronal activity in restrained and freely moving animals

A design of a compact micromanipulator giving possibility to introduce the microelectrode into unlimited depth of the brain and to realize the penetration of it into the narrow area of trepanized cranial of the animal several times has been worked out and described. Small size and mass of the design allow to set up several micromanipulators on the cranial of different experimental animals and to record the neuronal activity of the different brain structures simultaneously.     

Automatic recording of flea activity

An Insect Activity Monitor was created to measure the behavioural responses of fleas (Siphonaptera). The apparatus allows for a range of visual, chemo- and mechanoreceptor cues to be presented. The jumping response is detected by counting amplified pulses produced as the fleas land on a stretched membrane held over a microphone. Horizontal movements are detected using a system of infra-red beams and phototransistors which, when broken, are counted as a measure of activity. The apparatus was tested using Ceratophyllus hirundinis (Curtis), Ceratophyllus farreni (Rothschild) and Ceratophyllus rusticus (Wagner), co-inhabiting species from the nests of the house martin (Delichon u.urbica L.). No unaided emigration or immigration has been demonstrated in these species (Clark, 1988) and much of their time is spent confined to the darkness of the nest. In addition to species contrasts, females were more active than males and fleas were more active at 25 degrees C than at 18 degrees C. The more active bird flea Ceratophyllus garei (Rothschild) from the nest of pheasant (Phasianus colchicus L), was tested for responses to light of varying intensity. Activity was stimulated by white light, but not proportional to light intensity.     

Electronic recording of mosquito activity

Spontaneous locomotor activity of mosquitoes (Aedes aegypti) was tested over twenty-four hour periods using an electronic recording device which gave a permanent time graph of activity. Single mosquitoes were placed on a wire grid with alternate strands connected to the positive and negative poles of an electric circuit. Each time the mosquito moved, the electric current changed and the event was recorded by a pen-writer. The number of peaks per time interval gave the index of activity. Variables which may affect activity include age, physiological state, sex and strain. A distinct activity cycle was evident in both virgin and mated females but not in males; peak activity came in the early evening and activity was lowest in the early afternoon.

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Zusammenfassung Die spontane lokomotorische Aktivität von Mücken (Aedes aegypti) wurde über 24stündige Perioden mit Hilfe einer elektronischen Registriereinrichtung untersucht, die eine ununterbrochene Zeitschreibung der Aktivität ergab. Einzelne Mücken wurden auf einen Gürtel feiner Drähte gesetzt, deren Stränge abwechselnd zu den positiven und negativen Polen eines elektrischen Stromkreises führten. Jedesmal wenn sich die Mücke bewegte, änderte sich der elektrische Stromfluß; dieses Ereignis wurde von einer Schreibfeder aufgezeichnet. Die Anzahl der Ausschläge pro Zeiteinheit ergab den Aktivitätsindex. Variable, welche die Aktivität beeinflussen, umfassen Alter, physiologischen Zustand, Geschlecht und Abstammung. Bei jungfräulichen wie bei begatteten Weibchen war ein bestimmter Aktivitätszyklus erkennbar, jedoch nicht bei Männchen; der Aktivitätsgipfel lag in den frühen Abendstunden und die Aktivität war am zeitigen Nachmittag am geringsten.

     

Behavior-related neuronal reactions and dynamics of neuronal activity

Functionally, behavior-related discharges of associative neurons are an efferent flow of pulses continuously generated over the course of each behavioral act of an animal. However, predominant research approaches are based on the "stimulus - reaction" principle. Analysis of the dynamics of unit activity in monkeys during performance of a multi-step behavioral complex showed that differences related to different behavioral acts consist in composition changes in the active neurons (or their recombination) rather than in a number of responsive cells or involvement of action-specific neurons. Each combination of active neurons ensures the distribution of efferent signals characteristic of the given combination. These findings suggest the addressing coding of the efferent neuronal signals.     

Telemetric electrochemical sensor

A telemetric system was designed and constructed to sense pH and ethanol variation in aqueous solutions. The measured signals were transferred by software digitally and transmitted wirelessly by the telemeter, personal digital assistant (PDA), through the General Packet Radio Service (GPRS) protocol. The pH sensing electrode was designed to measure a chemical potential induced by a proton concentration gradient on the electrode's surface which exhibits internal Donnon diffusion behavior, and a linear relationship between the electrical potential and pH was found. The result shows that the wireless sensing system allowed not only long-term usage and long-distance transmission but also with high accuracy (e.g. S.D. less than +/-2%). The telemetric system can also be modified to measure ethanol concentration in aqueous solution amperometrically. It was found that the sensitivity of that ex situ measurements matched those of in field measurements with negligible deviation, less than 4%.     

Extracellular recording in neuronal networks with substrate integrated microelectrode arrays

A photolithographically produced array of 60 substrate-integrated microelectrodes was used for extracellular recording. Neuronal electrical activity was recorded from chicken retinal ganglion cells with or without stimulation by diffuse light. The retina was removed from chicken embryos of embryonic day 14–18. Only cells recorded from day 18 retina would react to photostimulation, increasing their activity when stimulated, corresponding to the developmental time course of photoreceptor differentiation.     

Fueling brain neuronal activity

The energy requirements of the brain are amazingly high. The brain represents about 2% of the body weight, but it receives 15% of the total blood flow provided by the cardiovascular system and consumes at least 25% circulating glucose plus 20% oxygen available in the body at rest. The cornerstone feature of the brain energy metabolism is its tight coupling with neuronal activity. An abnormality in the sequence of events allowing neurons to be adequately supplied with the necessary energy could have dramatic consequences exemplified in the neurodegenerative diseases such as epilepsy and Alzheimer’s disease. In this paper, we review the current views on the main pathways of neuronal energy supply.     

Alpha-chloralose suppression of neuronal activity

Alpha-chloralose, an anesthetic agent widely used in neurophysiologic studies, caused a significant and long-lasting suppression of single neuron activity recorded from two areas of the central nervous system in decerebrate cats. A 50 mg/kg dose (an average anesthetic dose used in many neurophysiologic studies) caused suppression of spontaneous and evoked activity of neurons in the dorsal horn of the spinal cord and greater suppression of neurons in the nucleus reticularis gigantocellularis (NRGC) of the medial medullary reticular formation. Many researchers are of the opinion that alpha-chloralose causes less suppression of the central nervous system (CNS) than other commonly used anesthetic agents. The neuronal suppression recorded in this study appears similar in many ways to suppression caused by other anesthetic agents in the same two areas of the CNS. The results of the present study suggest that alpha-chloralose may be capable of producing significant suppression of neurons in the dorsal horn of the spinal cord and NRGC. Its ability to influence other areas of the CNS should not be inferred from these results, but the data do indicate the importance of evaluating the effects of anesthetics upon neurophysiologic systems under study.     

A biotelemetry system recording fish activity

A biotelemetry system is described for obtaining, transmitting and recording the electromyograms (EMGs) produced in muscle activity of free-swimming fish as quantitative indicators of overall fish activity. The radiotransmitters used come in the form of cylindrical packages having two sensing electrodes, all fully implantable in the fish body cavity. EMGs are transmitted as radio pulses with the intensity of muscular activity determining the intervals between pulses. The packages also contain temperature sensors and fish temperatures are transmitted with every 32nd pulse. Transmitted EMG pulses are detected, 'measured' and stored by a single portable receiver (Model SRX_400, Lotek). Data can be subsequently transferred to a computer (which can also be portable) for storage, processing and statistical analysis. Transmitter battery life can be in excess of 7 months, permitting laboratory or field studies of long duration. Transmitter package implantation surgery requires a mid-ventral incision and internal securing of transmitter and sensing electrodes. Surgical silk, cyanoacrylate tissue adhesives, and polydioxanone (PD), a synthetic absorbable suture, were all tried as means of incision closure. The most effective was PD alone. Trials of the system consisted of forced swims by transmitter-equipped rainbow trout Oncorhynchus mykiss Walbaum. The data obtained provided an inverse linear relation between forced swim speed and EMG pulse interval. Trials were conducted at intervals over periods up to 2 months. Fish showed neither distress, nor difficulty in swimming up to maximum speeds of 60 cm s ⁻¹ (fish lengths 41.0, 44.4 cm).