In vitro characterisation results for O(2) reduction at Pt-based microelectrodes are presented an... more In vitro characterisation results for O(2) reduction at Pt-based microelectrodes are presented and compared with those for carbon-paste electrodes (CPEs). Cyclic voltammetry indicates a potential of -650 mV vs. SCE is required for cathodic reduction at both electrode types, and calibration experiments at this potential revealed a significantly higher sensitivity for Pt (-0.091 ± 0.006 μAmm(-2)μM(-1) vs. -0.048 ± 0.002 μAmm(-2)μM(-1) for CPEs). Since Pt electrodes are readily poisoned through contact with biological samples selected surface coated polymers (polyphenylenediamine (PPD), polymethyl methacrylate (PMMA) and Rhoplex(®)) were examined in biocompatibility studies performed in protein, lipid and brain tissue solutions. While small and comparable decreases in sensitivity were observed for bare Pt, Pt-Rhoplex and PMMA there was minimal change at the Pt-PPD modified electrode for each 24h treatment, including an extended 3 day exposure to brain tissue. The polymers themselves had no effect on the O(2) response characteristics. Further characterisation studies at the Pt-based microelectrodes confirmed interference free signals, no effect of pH and ion changes, and a comparable detection limit (0.08 ± 0.01 μM) and response time (<1 s) to CPEs. Although a significant temperature effect (ca. 3% change in signal for each 1 °C) was observed it is predicted that this will not be important for in vivo brain tissue O(2) measurements due to brain temperature homeostasis. These results suggest that amperometric Pt electrodes have the potential to be used reliably as an alternative to CPEs to monitor brain tissue O(2) over extended periods in freely-moving animals.
Amperometric sensors for oxygen and glucose allow for real time recording from the brain in freel... more Amperometric sensors for oxygen and glucose allow for real time recording from the brain in freely-moving animals. These sensors have been used to detect activity- and drug-induced changes in metabolism in a number of brain regions but little attention has been given over to the hippocampus despite its importance in cognition and disease. Sensors for oxygen and glucose were co-implanted into the hippocampus and allowed to record for several days. Baseline recordings show that basal concentrations of hippocampal oxygen and glucose are 100.26±5.76 μM and 0.60±0.06 mM respectively. Furthermore, stress-induced changes in neural activity have been shown to significantly alter concentrations of both analytes in the hippocampus. Administration of O2 gas to the animals' snouts results in significant increases in hippocampal oxygen and glucose and administration of N2 gas results in a significant decrease in hippocampal oxygen. Chloral hydrate-induced anaesthesia causes a significant increase in hippocampal oxygen whereas treatment with the carbonic anhydrase inhibitor acetazolamide significantly increases hippocampal oxygen and glucose. These findings provide real time electrochemical data for the hippocampus which has been previously impossible with traditional methods such as microdialysis or ex vivo analysis. As such, these sensors provide a window into hippocampal function which can be used in conjunction with behavioural and pharmacological interventions to further elucidate the functions and mechanisms of action of the hippocampus in normal and disease states.
The hippocampus plays a vital role in learning and memory and is susceptible to damage following ... more The hippocampus plays a vital role in learning and memory and is susceptible to damage following hypoglycaemic shock. The effect of an acute administration of insulin on hippocampal function has been described in terms of behavioural deficits but its effect on hippocampal oxygen and glucose is unclear. Glucose oxidase biosensors (detecting glucose) and carbon paste electrodes (detecting oxygen) were implanted into the hippocampus of Sprague Dawley rats. Animals were allowed to recover and real-time recordings were made in order to determine the effects of fasting, insulin administration (15U/kg; i.p.) and reintroduction of food on hippocampal oxygen and glucose. Fasting caused a significant decrease in hippocampal glucose over the course of 24h. Insulin administration produced a significant decrease in hippocampal glucose along with a significant increase in hippocampal oxygen. Finally, the reintroduction of food resulted in glucose levels significantly increasing along with a trans...
In vitro characterisation results for O(2) reduction at Pt-based microelectrodes are presented an... more In vitro characterisation results for O(2) reduction at Pt-based microelectrodes are presented and compared with those for carbon-paste electrodes (CPEs). Cyclic voltammetry indicates a potential of -650 mV vs. SCE is required for cathodic reduction at both electrode types, and calibration experiments at this potential revealed a significantly higher sensitivity for Pt (-0.091 ± 0.006 μAmm(-2)μM(-1) vs. -0.048 ± 0.002 μAmm(-2)μM(-1) for CPEs). Since Pt electrodes are readily poisoned through contact with biological samples selected surface coated polymers (polyphenylenediamine (PPD), polymethyl methacrylate (PMMA) and Rhoplex(®)) were examined in biocompatibility studies performed in protein, lipid and brain tissue solutions. While small and comparable decreases in sensitivity were observed for bare Pt, Pt-Rhoplex and PMMA there was minimal change at the Pt-PPD modified electrode for each 24h treatment, including an extended 3 day exposure to brain tissue. The polymers themselves had no effect on the O(2) response characteristics. Further characterisation studies at the Pt-based microelectrodes confirmed interference free signals, no effect of pH and ion changes, and a comparable detection limit (0.08 ± 0.01 μM) and response time (<1 s) to CPEs. Although a significant temperature effect (ca. 3% change in signal for each 1 °C) was observed it is predicted that this will not be important for in vivo brain tissue O(2) measurements due to brain temperature homeostasis. These results suggest that amperometric Pt electrodes have the potential to be used reliably as an alternative to CPEs to monitor brain tissue O(2) over extended periods in freely-moving animals.
Amperometric sensors for oxygen and glucose allow for real time recording from the brain in freel... more Amperometric sensors for oxygen and glucose allow for real time recording from the brain in freely-moving animals. These sensors have been used to detect activity- and drug-induced changes in metabolism in a number of brain regions but little attention has been given over to the hippocampus despite its importance in cognition and disease. Sensors for oxygen and glucose were co-implanted into the hippocampus and allowed to record for several days. Baseline recordings show that basal concentrations of hippocampal oxygen and glucose are 100.26±5.76 μM and 0.60±0.06 mM respectively. Furthermore, stress-induced changes in neural activity have been shown to significantly alter concentrations of both analytes in the hippocampus. Administration of O2 gas to the animals' snouts results in significant increases in hippocampal oxygen and glucose and administration of N2 gas results in a significant decrease in hippocampal oxygen. Chloral hydrate-induced anaesthesia causes a significant increase in hippocampal oxygen whereas treatment with the carbonic anhydrase inhibitor acetazolamide significantly increases hippocampal oxygen and glucose. These findings provide real time electrochemical data for the hippocampus which has been previously impossible with traditional methods such as microdialysis or ex vivo analysis. As such, these sensors provide a window into hippocampal function which can be used in conjunction with behavioural and pharmacological interventions to further elucidate the functions and mechanisms of action of the hippocampus in normal and disease states.
The hippocampus plays a vital role in learning and memory and is susceptible to damage following ... more The hippocampus plays a vital role in learning and memory and is susceptible to damage following hypoglycaemic shock. The effect of an acute administration of insulin on hippocampal function has been described in terms of behavioural deficits but its effect on hippocampal oxygen and glucose is unclear. Glucose oxidase biosensors (detecting glucose) and carbon paste electrodes (detecting oxygen) were implanted into the hippocampus of Sprague Dawley rats. Animals were allowed to recover and real-time recordings were made in order to determine the effects of fasting, insulin administration (15U/kg; i.p.) and reintroduction of food on hippocampal oxygen and glucose. Fasting caused a significant decrease in hippocampal glucose over the course of 24h. Insulin administration produced a significant decrease in hippocampal glucose along with a significant increase in hippocampal oxygen. Finally, the reintroduction of food resulted in glucose levels significantly increasing along with a trans...
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