Journal of Neuroscience Methods
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Shape, connectedness and dynamics in neuronal networks.
The morphology of neurons is directly related to several aspects of the nervous system, including its connectedness, health, development, evolution, dynamics and, ultimately, behavior. Such interplays of the neuronal morphology can be understood within the more general shape-function paradigm. The c...
Water-tight membranes from neuronal morphology files.
We present an algorithm to form watertight 3D surfaces consistent with the point-and-diameter based neuronal morphology descriptions widely used with spatial electrophysiology simulators. Such morphology descriptions are readily available online and may come from light-microscopy tracings or from an...
Whole brain monoamine detection and manipulation in a stalk-eyed fly.
We developed a dissection and sample preparation method sufficiently sensitive to measure monoamine concentrations from whole brain samples of small insects. This new method allows the detection of monoamines from a single fly brain using high performance liquid chromatography with electrochemical d...
FACS binding assay for analysing GDNF interactions.
We developed a FACS-based binding assay for GDNF as an alternative to current methods. We demonstrated that the FACS-based assay using TGW cells allowed readily detection of GDNF binding and displacement to endogenous receptors. The dissociation constant and half maximal inhibitory concentration obt...
A simple and fast method for tissue cryohomogenization enabling multifarious molecular extraction.
We refined the methods for cryohomogenization to allow efficient use of a single experimental sample so that it can easily be divided into fractions for extraction of different molecular components, immediately or after storage. Using western blot, nanodrop UV/V spectrometry, and a bioanalyzer, we s...
A magnetic resonance imaging-safe method for the study of human eyeblink conditioning.
Eyeblink conditioning (EBC) is a widely used translational probe of cerebellar function in both humans and non-human animals. Decades of animal research have identified the cerebellum as critical for EBC. While there is evidence for the involvement of the cerebellum in human EBC, the neural circuitr...
Delayed repair of the peripheral nerve: A novel model in the rat sciatic nerve.
Peripheral nerve reconstruction is seldom done in the acute phase of nerve injury due to concomitant injuries and the uncertainty of the extent of nerve damage. A proper model that mimics true clinical scenarios is critical but lacking. The aim of this study is to develop a standardized, delayed sci...
Simple generation of neurons from human embryonic stem cells using agarose multiwell dishes.
We describe here a protocol based on commercially available reusable silicone micromolds and two small molecule growth factor inhibitors to simply and reproducibly generate human neurons from hESC. Hundreds of neurospheres were generated with a single pipettation of hESC into agarose multiwell plate...
A simple method for isolating rat brain mitochondria with high metabolic activity: Effects of EDTA and EGTA.
Our results showed alterations in both active and resting respiration, which were dependent on both the addition of EDTA or EGTA to the isolation buffer and the chelator concentration used. However, the addition of chelator to the isolation medium does not modify the mitochondria structure as assess...
Membrane configuration optimization for a murine in vitro blood-brain barrier model.
A powerful experimental tool used to study the dynamic functions of the blood-brain barrier (BBB) is an in vitro cellular based system utilizing cell culture inserts in multi-well plates. Currently, usage of divergent model configurations without explanation of selected variable set points renders d...