Role of vascular endothelial growth factor as a critical neurotrophic factor for the survival and physiology of motoneurons

Vascular endothelial growth factor(VEGF)was discovered by its angiogenic activity.However,during evolution,it appeared earlier as a neurotrophic factor required for the development of the nervous system in invertebrates lacking a circulatory system.We aimed at reviewing recent evidence indicating that VEGF has neuroprotective effects in neurons exposed to a variety of insults.Of particular interest is the link established between VEGF and motoneurons,especially after the design of the VEGFδ/δmutant mice.These mice are characterized by low levels of VEGF and develop muscle weakness and motoneuron degeneration resembling amyotrophic lateral sclerosis.The administration of VEGF through several routes to animal models of amyotrophic lateral sclerosis delays motor impairment and motoneuron degeneration and increases life expectancy.There are new recent advances in the role of VEGF in the physiology of motoneurons.Our experimental aims use the extraocular(abducens)motoneurons lesioned by axotomy as a model for studying VEGF actions.Axotomized abducens motoneurons exhibit severe alterations in their discharge activity and a loss of synaptic boutons.The exogenous administration of VEGF to axotomized abducens motoneurons,either from the transected nerve or intraventricularly,fully restores the synaptic and discharge properties of abducens motoneurons,despite being axotomized.In addition,when an anti-VEGF neutralizing antibody is delivered from the muscle to intact,uninjured abducens motoneurons,these cells display alterations in their discharge pattern and a loss of synaptic boutons that resemble the state of axotomy.All these data indicate that VEGF is an essential neurotrophic factor for motoneurons.

Microelectrode array for bioelectrical signal stimulation and recording

A microelectrode array（MEA） is presented, which is composed of 60 independent electrodes with 59 working ones and one reference one, and they are divided into 30 pairs. Except for the reference electrode, each pair consists of one stimulating electrode and one recording electrode. Supported by the peripheral circuits, four electrode states to study the bioelectrical signal of biological tissue or slice cultured in-vitro on the surface of the electrodes can be realized through each pair of electrodes. The four electrode states are stimulation, recording, stimulation and recording simultaneously, and isolation. The state of each pair of working electrodes can be arbitrarily controlled according to actual needs. The MEAs are fabricated in printed circuit board （PCB） technology. The total area of the PCB-based MEA is 49 mm × 49 mm. The impedance measurement of MEA is carried out in 0.9% sodium chloride solution at room temperature by means of 2-point measurements with an Agilent LCR meter, and the test signal for the impedance measurement is sinusoidal （AC voltage 50 mV, sweeping frequency 20 Hz to 10 kHz）. The electrode impedance is between 200 and 3 kΩ while the frequency is between 500 and 1 000 Hz. The electrode impedance magnitude is inversely proportional to the frequency. Experiments of toad sciatic nerve in-vitro stimulation and recording and signal regeneration between isolated toad sciatic nerves are carried out on the PCB-based MEA. The results show that the MEA can be used for bioelectrical signal stimulation, recording, stimulation and recording simultaneously, and isolation of biological tissues or slices in-vitro.

A novel implantable multichannel silicon-based microelectrode

Silicon-based microelectrodes have been confirmed to be helpful in neural prostheses. The fabricated 7-channel silicon-based microelectrode was feasible to be implanted into the brain cortex. The manufacturing process by micro-electromechanical system （MEMS） technology was detailed with four photolithographic masks. The microscopic photographs and SEM images indicated that the probe shank was 3 mm long, 100μm wide and 20 μm thick with the recording sites spaced 120μm apart for good signal isolation. To facilitate the insertion and minimize the trauma, the microelectrode is narrowed down gradually near the tip with the tip taper angle of 6 degrees. Curve of the single recording site impedance versus frequency was shown by test in vitro and the impedance declined from 150.5 kΩ to 6.0 kΩ with frequency changing from 10 k to 10 MHz.

Automatic Spike Sorting Based on Robust Clustering

The collected spikes from extracellular recordings usually contain noisy data and outliers, which make it difficult to separate them. A method for spike sorting based on robust clustering is proposed to deal with the problem. The clustering method combines the advantage of fuzzy clustering and robust statistical estimators. The number of dusters is obtained by fuzzy cluster validity. In order to reduce the influence of outliers, the validity index is calculated using the weighting intra-cluster distances. The proposed method is suitable to separate neural spikes in the presence of noisy data and outfiers. The experiment on real data shows its performance.