Biomedical subjects

Yannan Ouyang

Publications and source records attributed to Yannan Ouyang.

5 recordsLinked to original sources

PubMed2005

Cooling blocks rat hippocampal neurotransmission by a presynaptic mechanism: observations using 2-photon microscopy.

Over the past decade there has been great interest in the therapeutic potential of brain cooling for epilepsy, stroke, asphyxia and other neurological diseases. However, there is still no consensus regarding the neurophysiological effect(s) of brain cooling. We employed standard physiological techniques and 2-photon microscopy to directly examine the effect of temperature on evoked neurotransmitter release in rat hippocampal slices. We observed a monotonic decline in extracellular synaptic potentials and their initial slope over the temperature range 33-20 degrees C, when the slices were cooled to a new set point in less than 5 s. Imaging the fluorescent synaptic marker FM1-43 with 2-photon microscopy showed that the same cooling protocol dramatically reduced transmitter release between 33 and 20 degrees C. Cooling also reduced the terminal FM1-43 destaining that was induced by direct depolarization with elevated K+, indicating that axonal conduction block cannot account for our observations. The temperature dependence of FM1-43 destaining correlated well with the effect of temperature on field potential slope, compatible with a presynaptic explanation for our electrophysiological observations. Optical measurement of FM1-43 dissociation from cell membranes was not affected by temperature, and rapid cooling of slices loaded with FM1-43 did not increase their fluorescence. Our experiments provide visible evidence that a major neurophysiological effect of cooling in the mammalian brain is a reduction in the efficacy of neurotransmitter release. This presynaptic effect may account for some of the therapeutic benefits of cooling in epilepsy and possibly stroke.

Animals↗

PubMed2005

In vivo imaging of dendritic spines during electrographic seizures.

Epilepsy is associated with significant neurological morbidity, including learning disabilities, motor deficits, and behavioral problems. Although the causes of neurological dysfunction in epilepsy are multifactorial, accumulating evidence indicates that seizures in themselves may directly cause brain injury. Although it is clear that seizures can result in neuronal death, it is likely that under some circumstances seizures can induce more subtle functional or structural alterations in neurons. We induced focal neocortical seizures with 4-aminopyridine in transgenic mice expressing green fluorescent protein in cortical neurons and sequentially imaged individual dendrites in living animals with two-photon laser-scanning microscopy to determine whether these seizures caused acute alterations in dendritic spine morphology. No dendritic alterations were observed in anesthetized animals during electrographic seizures over a 3-hour period. Similarly, in unanesthetized mice, low-stage, clinical electrographic seizures had minimal effect on dendritic spines. More severe, high-stage seizures in unanesthetized mice were associated with a moderate loss of spines and dendritic swelling, but this effect may have been contingent on a synergistic action of phototoxicity from the imaging method itself. Overall, our results suggest that most neocortical seizures have minimal acute effects on dendrites over several hours, but may predispose to dendritic injury under extreme conditions.

4-Aminopyridine↗

PubMed2005

Transient decrease in F-actin may be necessary for translocation of proteins into dendritic spines.

It remains poorly understood as to how newly synthesized proteins that are required to act at specific synapses are translocated into only selected subsets of potentiated dendritic spines. Here, we report that F-actin, a major component of the skeletal structure of dendritic spines, may contribute to the regulation of synaptic specificity of protein translocation. We found that the stabilization of F-actin blocked the translocation of GFP-CaMKII and inhibited the diffusion of 3-kDa dextran into spines (in 2-3 weeks cultures). Neuronal activation in hippocampal slices and cultured neurons led to an increase in the activation (decrease in the phosphorylation) of the actin depolymerization factor, cofilin, and a decrease in F-actin. Furthermore, the induction of long-term potentiation by tetanic stimulation induced local transient depolymerization of F-actin both in vivo and in hippocampal slices (8-10 weeks), and this local F-actin depolymerization was blocked by APV, a N-methyl-D-aspartate (NMDA) receptor antagonist. These results suggest that F-actin may play a role in synaptic specificity by allowing protein translocation into only potentiated spines, gated through its depolymerization, which is probably triggered by the activation of NMDA receptors.

2-Amino-5-phosphonovalerate↗

PubMed2004

Activity-dependent transcription regulation of PSD-95 by neuregulin-1 and Eos.

Neuregulin-1 (Nrg-1) contains an intracellular domain (Nrg-ICD) that translocates into the nucleus, where it may regulate gene expression upon neuronal depolarization. However, the identity of its target promoters and the mechanisms by which it regulates transcription have been elusive. Here we report that, in the mouse cochlea, synaptic activity increases the level of nuclear Nrg-ICD and upregulates postsynaptic density protein-95 (PSD-95), a scaffolding protein that is enriched in post-synaptic structures. Nrg-ICD enhances the transcriptional activity of the PSD-95 promoter by binding to a zinc-finger transcription factor, Eos. The Nrg-ICD-Eos complex induces endogenous PSD-95 expression in vivo through a signaling pathway that is mostly independent of gamma-secretase regulation. This upregulation of PSD-95 expression by the Nrg-ICD-Eos complex provides a molecular basis for activity-dependent synaptic plasticity.

Acoustic Stimulation↗

PubMed2003

Optical recordings of Ca2+ signaling activities from identified inner ear cells in cochlear slices and hemicochleae.

One of the major obstacles hindering the progress of studies on mammalian cochlear physiology is the inaccessibility of inner ear cells located in a complex structure of the bony labyrinth. We describe here a technique to record cellular fluorescent signals from any identified inner ear cells in cochlear slices and hemicochleae. Cochlear slices were obtained from postnatal rats (P0-P7) before the cochlea completely ossify, and hemicochleae were cut from older animals (P7-adult). Individual inner ear cells were visually identified using infrared differential interference contrast or oblique illumination optics. Techniques were developed for either bulk-loading cells or loading selected single cells with Ca(2+) indicator dyes, and for maintaining functional viability of cochlear slices/hemicochleae for recordings. Robust and reliable responses of ligand-gated receptors were recorded from individual inner ear cells (e.g. hair cells, spiral ganglion neurons etc.) for at least 24 h after slices/hemicochleae were cut by an oscillating tissue slicer. The technique described here allowed direct observations of [Ca(2+)](i) activities from multiple cells simultaneously in situ, thus providing a feasible way to study the intercellular communication or networking activities from identified cells in the inner ear.

Action Potentials↗