Biomedical subjects

Alan R Kay

Publications and source records attributed to Alan R Kay.

5 recordsLinked to original sources

PubMed2006

A startling role for synaptic zinc.

It has long been known that the synaptic vesicles of certain glutamatergic terminals, as well as some inhibitory terminals, are richly supplied with zinc ions, yet the functional role of this pool of zinc in synaptic transmission has remained elusive. In this issue of Neuron, Hirzel et al. provide direct in vivo evidence that endogenous zinc is required for proper functioning of neuronal circuitry in the brainstem and spinal cord. They show that knockin mice carrying a point mutation which eliminates zinc potentiation of alpha1-containing glycine receptors develop severe sensorimotor deficits characteristic of impaired glycinergic neurotransmission.

Animals↗

PubMed2006

Imaging synaptic zinc: promises and perils.

It is well established that some excitatory nerve terminals have high concentrations of Zn(2+) in their synaptic vesicles. For some time, it has been believed that synaptic Zn(2+) is released during neurotransmission and acts as a neuromodulator. Fluorescent Zn(2+) indicators that do not penetrate membranes offer the prospect of rendering the release of Zn(2+) visible. Here, I take a critical look at fluorimetric imaging experiments devised to determine whether Zn(2+) is released and show that they are particularly susceptible to artifacts. Moreover, I will argue that recent experiments suggest that, rather than being released, Zn(2+) is presented to the extracellular space firmly coordinated to presynaptic macromolecules.

Animals↗

PubMed2005

Influence of location of a fluorescent zinc probe in brain slices on its response to synaptic activation.

The precise role of the high concentration of ionic zinc found in the synaptic vesicles of certain glutamatergic terminals is unknown. Fluorescent probes with their ability to detect ions at low concentrations provide a powerful approach to monitoring cellular Zn2+ levels. In the last few years, a number of fluorescent probes (indicators) have been synthesized that can be used to visualize Zn2+ in live cells. The interpretation of data gathered using such probes depends crucially on the location of the probe. Using acutely prepared hippocampal slices, we provide evidence that the Zn2+ probes, ZnAF-2 and ZP4, are membrane permeant and are able to pass into synaptic vesicles. In addition, we show that changes in fluorescence of the Zn2+ probes can be used to monitor presynaptic activity; however, these changes are inconsistent with Zn2+ release.

Action Potentials↗

PubMed2004

Detecting and minimizing zinc contamination in physiological solutions.

BACKGROUND: To explore the role of zinc (Zn) in cellular physiology it is important to be able to control and quantify the level of Zn contamination in experimental solutions. A technique that relies on a Zn-sensitive fluorimetric probe is introduced for measuring Zn concentrations as low as 100 pM. The method depends on the combination of the Zn-probe FluoZin-3 together with a slow Zn-chelator, Ca-EDTA, that reduces the background Zn levels and allows repeated measurements in the same solution. RESULTS: The method was used to determine which common labware items could leach Zn into solution. Contamination was predictably found to arise from stainless steel and glass. Perhaps less expectedly it was also introduced by methacrylate cuvettes, plastic tissue culture dishes and other plastic labware. The release of nickel from stainless steel electrodes was also imaged using the fluorescent probe Newport Green. CONCLUSION: Zn contamination may arise from rather unexpected sources; it is important that all aspects and components used in the course of an experiment be analyzed for the possibility of introducing contaminants.

Electrodes↗

PubMed2003

Evidence for chelatable zinc in the extracellular space of the hippocampus, but little evidence for synaptic release of Zn.

Zinc colocalizes with glutamate in the synaptic vesicles of certain glutamatergic vesicles in the mammalian brain. Here, I introduce a method for detecting Zn in the extracellular space of brain slices and another method for detecting the passage of Zn out of the slice. In both cases, the fluorimetric Zn probe FluoZin-3 is used in conjunction with a slow Zn chelator, Ca-EDTA, to reduce background fluorescence. In addition, a new Zn chelator, ethylenediiminodi-2-pentanedioic acid, with little affinity for Ca or Mg is introduced. These tools are then used to show that little Zn (approximately 2 nm) is released during the course of synaptic transmission into the extracellular space. However, when hippocampal slices are subjected to a high potassium stimulus (50 mM) combined with an increase in osmolarity, Zn is externalized in the Timm's-stained areas (approximately 6 nm). This stimulus also leads to even greater Zn elevations in area CA1 that is only weakly stained by the Timm's method. Nevertheless, even under these conditions, little if any Zn makes its way out of the slices. I present evidence for a layer of Zn in the extracellular space that maps onto the Timm's stained region of the hippocampus. This Zn veneer appears to be loosely associated with molecules in the extracellular space and may be the raison d'être for vesicular Zn.

Animals↗