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Marking the tip location of PO2 microelectrodes or glass micropipettes.

The necessity of localizing the PO2 recording sites in the carotid body of cats has led us to modify our O2 microelectrode to be able to mark the position of the tip. Gold is sputtered on the electrode, and then iron is plated on the gold. The carotid body of cats was exposed, and PO2 was recorded at different depths in the body after which iron was deposited. At the end of the experiment, the excised body was placed in a 1:1 mixture of 4% potassium ferrocyanide and 4% acetic acid for 1 h or more. After that, the body was fixed in a 1:1 mixture of 40% formaldehyde and 95% ethyl alcohol for 1-2 days. Paraffin sections cut at 10 micrometer revealed the recording sites as bright blue spots. Since the cathode size can be made as small as 1 micrometer, discrete spots (a few microns or larger depending on current and time) can be produced at the PO2 recording sites. Several locations can be marked in a few minutes without damage to the tissue or the electrode. In 21 cats all of the 62 marked spots were found. In 16 peripheral (depth 50-150 micrometers) locations the PO2 averaged 101 Torr and in 46 deep locations 73 Torr. This technique is applicable to glass micropipettes and is a unique tool in correlating the structure and function of various tissues.

Animals↗

Membrane properties of dentate gyrus granule cells: comparison of sharp microelectrode and whole-cell recordings.

1. Whole-cell and sharp electrode recordings from adult rat dentate gyrus GCs were performed in the 400-microns-thick hippocampal slice preparation maintained at 34 +/- 1 degrees C. Intrinsic membrane properties of granule cells (GCs) were evaluated with the use of a switching current-clamp amplifier. 2. With the whole-cell technique, the average resting membrane potential (RMP) of GCs was -85 mV when a potassium gluconate electrode solution was used versus -74 mV measured with potassium acetate-filled sharp microelectrodes. The membrane voltage response to injected current was linear over two membrane potential ranges, greater than 10 mV hyperpolarized from RMP and between 10 mV more negative than RMP and -62 mV. The average input resistances (RN) calculated over these ranges were 107 and 228 M omega in the whole-cell recordings versus 37 and 54 M omega in the sharp electrode recordings. There was no correlation between RMP and RN with either recording technique. The membrane time constant (tau m) determined at the RMP was 26.9 ms for whole-cell recordings and 13.9 ms for sharp electrode recordings. 3. There was no evidence of time-dependent changes in RMP, RN, and tau m in whole-cell recordings, although the slow inward rectification seen at hyperpolarized potentials decreased over 30-60 min. Addition of calcium buffers to the whole-cell recording solution did not result in a significant change in the average RMP, the average RN, or the average tau m. 4. Action potential threshold was comparable in whole-cell (-49 mV) and sharp electrode (-52 mV) recordings, but action potential amplitude was larger in whole-cell (126 mV) than in sharp electrode (106 mV) recordings. Spike frequency adaptation was present in the whole-cell recordings and could be abolished by addition of calcium buffers to the electrode solution. 5. We estimated rho, the ratio of dendritic to somatic conductance, to be 5.1 for the whole-cell records and 2.1 for sharp electrode recordings. The electrotonic length of the equivalent cylinder representing the cell processes was estimated to be 0.49 from the whole-cell data and 0.79 from the sharp electrode recordings. This implies that at rest there is only a 10% decrement in steady-state membrane voltage along the length of the dendrite due to shunting across the membrane resistance; small synaptic events occurring in the distal dendritic tree will therefore have a more substantial influence on the soma than previous analyses suggested.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

An examination of frog myelinated axons using intracellular microelectrode recording: the role of voltage-dependent and leak conductances on the steady-state electrical properties.

1. Intracellular microelectrode current-clamp technique was used to study the steady-state membrane properties of single intact large primary afferent axons (conduction velocity > 10 m/s) attached to isolated hemisected frog spinal cord. 2. Hyperpolarizing electrotonic potentials (ETPs) had a slow complex multiphasic charging. This complex charging could be approximated by two time constants: one in the range of 70-210 ms, the other of < 20 ms. 3. Two regions of outward and inward rectification hyperpolarized to the resting membrane potential were observed, in addition to the previously characterized outward rectification active at potentials depolarized to resting membrane potential. The peak and steady-state input resistance of these axons in tetrodotoxin Ringer solution was on average 65.6 +/- 21.1 and 31.1 +/- 10.8 M omega, mean +/- SE, respectively. 4. Application of external tetraethylammonium (10-20 mM) significantly depolarized the axon and decreased the outward rectification just hyperpolarized to the resting membrane potential. This outward rectification could also be blocked by external barium ions (2-10 mM). 5. Activation of an inward or anomalous rectification in these axons was observed 300-600 ms after the start of a current pulse. In addition, a depolarizing afterpotential (DAP) (1-3 mV in amplitude, 500 ms-10 s in duration) was evident after a current pulse in which inward rectification had been activated. This DAP most likely reflected the slow inactivation of the inwardly rectifying conductance. 6. Inward rectification was blocked by external application of cesium ions (1-3 mM) but it was insensitive to external application of barium ions (2-10 mM). The blockade of the voltage attenuation was accompanied by a disappearance of the DAP and an increase in the charging time constant of the axon. This blockade resulted in a single linear voltage-current (V-I) relationship. Axons now had, on average, an input resistance of 114 +/- 19.1 M omega. 7. Reducing the concentration of external potassium ions increased both the peak and steady-state slope resistance. Reducing the external sodium concentration altered the ETPs and the V-I relationship little but it consistently reduced the magnitude and length of the DAP. These results are compatible with the hypothesis that anomalous rectification is a mixed ionic conductance dependent on potassium and sodium ions in the external media. 8. Overall, the V-I relationship of these intact axons had both linear and nonlinear regions reflecting the activity of numerous slowly activating and inactivating conductances. (ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

Intraoperative microelectrode recording equipment: what features are necessary?

Intraoperative neurophysiologic methods for localizing targets deep in the brain require the use of specialized monitoring and recording equipment, including stimulators, neurophysiologic recording devices, and image manipulation tools. When using microelectrode recording devices there are some specifications that are more important than others, such as signal-to-noise ratios and amplifier impedance. As more companies develop tools to be used in the operating room, the end users have more choices. Some of the more important specifications are discussed and a comparison is made of the five major brands on the market today.

Action Potentials↗

Safety and risk of microelectrode recording in surgery for movement disorders.

There is an ongoing controversy about whether it is necessary to use microelectrode recording (MER) techniques in stereotactic surgery for Parkinson's disease and other movement disorders. This paper consists of a critical review of the published literature in order to analyze the value of MER in providing safe, efficient and accurate functional stereotactic surgery. Review of the literature revealed that MER techniques do not necessarily improve targeting accuracy or clinical results, compared to techniques using impedance monitoring and macrostimulation. In terms of safety for the patients, however, MER techniques are relatively safe, but non-Mer techniques, based on macrostimulation-guided surgery, are at least five times safer.

Globus Pallidus↗

Variability in position of the subthalamic nucleus targeted by magnetic resonance imaging and microelectrode recordings as compared to atlas co-ordinates.

BACKGROUND: Traditional methods for localisation of target nuclei for deep brain stimulation (DBS) have used brain atlas co-ordinates for initial targeting. It is now possible to visualise the subthalamic nucleus (STN) on magnetic resonance imaging (MRI) and determine the individual variability of its position. METHODS: The present study was performed in patients undergoing STN DBS for Parkinson's disease. The STN was directly targeted from axially obtained MRI and verified with microelectrode recordings. Postoperatively, the most effective contact was identified for each patient, and its position was calculated. RESULTS: Fifty electrodes were inserted in 25 patients. The target position varied considerably in relation to the mid-commissural point. The mean effective contact position lies just dorsal to the location of the STN in a standard brain atlas. CONCLUSION: The STN varies in position, and can be accurately targeted from MRI alone.

Adult↗

Long-term follow-up study of nucleus ventralis intermedius and ventrolateralis thalamotomy using a microelectrode technique in parkinsonism.

Long-term observation over 3-8 postoperative years of cases of Parkinson disease operated by stereotactic thalamotomy using a microelectrode recording technique is reported. The procedure is specifically useful in the following four groups: (1) tremor-dominant cases, (2) hemiparkinsonism, (3) cases with marked asymmetry in motor symptoms and (4) juvenile parkinsonism presenting levodopa-induced dyskinesia.

Follow-Up Studies↗

Thalamic microelectrode recording and microstimulation in central and deafferentation pain.

Microelectrode recording and microstimulation (in 1 case macrostimulation) were used to localize stereotactic targets in 8 patients with central and deafferentation pain and 1 'deafferented control' patient without pain. Off-line study revealed two features commonly proposed as potential physiological factors for such pain, but they were present in both pain and control patients--somatotopographic reorganization and the presence of bursting cells. In all patients in whom stimulation induced pain referred to deafferented parts of the contralateral body, hyperpathia or allodynia were present instead of normal paraesthetic, sensorimotor or thermal responses.

Electric Stimulation↗

Microelectrode recording for the somatotopic placement of stereotactic thalamic lesions in the treatment of parkinsonian and cerebellar intention tremor.

Patients with parkinsonian tremor and rigidity and cerebellar intention tremor can be effectively treated with thalamic lesions provided that an accurate neurophysiologic method is employed to compensate for individual spatial variability of subcortical structures. The method described employs a technique of microelectrode recording in which the homuncular organization in ventralis posterior is explored and gives information as to the ideal laterality of lateral thalamic nuclear mass lesions for the treatment of tremor. This procedure has been used in 11 patients, 9 with Parkinson's disease and 2 with cerebellar intention tremor with good results and no complications to date.

Cerebellar Diseases↗

Pacemaker activity of the pelvicalyceal border recorded by an intracellular glass microelectrode.

Applying intracellular glass microelectrodes to the smooth muscle obtained from the guinea pig pelvicalyceal border, we have succeeded in recording periodic spontaneous action potentials, the so-called slow waves, which are characteristic of a pacemaker. This experiment proved that the pacemaker exists at the pelvicalyceal border. The slow waves originate from the pelvicalyceal border, close to the papilla renalis; they show a biphasic pattern and have the characteristics of a spontaneously excitable cell. The resting potential is 42.3 +/- 1.1 mV, the spontaneous depolarizing potential is 12.1 +/- 0.7 mV, and the depolarizing period is 12/min. The rate of rise of the potential is 22.9 +/- 2.1 mV/s in the first slow depolarization phase, and 50.9 +/- 9.5 mV/s in the following rapid depolarization phase. The rate of decline of the potential is 47.3 +/- 5.4 mV/s in the repolarization phase.

Action Potentials↗

Electrochemical profile of K and Cl ions across the proximal tubule of bullfrog kidneys: a study using double-barreled ion-sensitive microelectrodes.

Micropuncture study was performed in the proximal segment of bullfrog nephrons with double-barreled ion-sensitive microelectrodes to determine the electrochemical profile of K and Cl across the individual borders of tubular epithelium. The mean peritubular potential obtained was -68.4 mV, and the K activity in the plasma and the cell interior was 2.64 and 61.6 mEq/liter, respectively. The calculated equilibrium potential for K was 79.3 mV. Therefore, the cell K must be maintained by some K uptake mechanism against an electrochemical gradient of 11 mV. Further, the K activity of the tubular fluid was 2.92 mEq/liter and the actually measured PD across the brush border membrane was -55.7 mV. Since the calculated Nernst K potential is 76.8 mV, the K entry from lumen to cell must be done against an electrochemical gradient of 21.1 mV in the net. These facts might suggest a possibility of bilateral existence of K uptake mechanism in both the peritubular and brush border membranes of proximal tubular cells. In contrast, the Cl activity was measured to be 9.30, 78.0, and 72.5 mEq/liter for the cell, tubular fluid, and plasma, respectively. The Nernst Cl potentials calculated were 53.6 and 51.7 mV for the brush border and the peritubular membrane, the data being less than the actually determined values, 55.7 and 68.4 mV, respectively. Thus, the Cl ion seemed to distribute passively throughout its reabsorptive process in the bullfrog proximal tubule.

Animals↗

The effect of ouabain on the isolated sinus node preparation of the rabbit studied with microelectrodes.

In 30 isolated, spontaneously beating right atrial preparations of the rabbit, the arrhythmogenic actions of ouabain were studied with microelectrodes. Ouabain (10(-6) M) uniformly produced the following events: (1) a gradual increase in sinus rate with periods of alternation of beta-to-beat atrial intervals, (2) a rapid rate and regular rhythm, (3) severe irregularity of rhythm, (4) total atrial arrest. Cycle length decreased from 394 msec +/- 43 (mean +/- SD) during control to 229 msec +/- 26 during the period of rapid rate and regular rhythm (P less than 0.001). Pretreatment with atropine or propranolol did not abolish this positive chronotropic action of ouabain. Maps of the spread of activation in the area of the sinus node in eight experiments revealed that the speeding up of rate is accompanied by a shift of dominant pacemaker site toward the sinoatrial border. In explanation, it is shown that sinoatrial border fibers develop strong diastolic depolarization, whereas dominant pacemaker fibers do so to a lesser extent or not at all. The atrial arrhythmias--best to be described as "digitalis-induced sinus tachycardia"--may represent the experimental counterpart of the so-called "paroxysmal atrial tachycardia with block" described as a common manifestation of digitalis intoxication in man.

Action Potentials↗

Cumulative depletions of extracellular calcium in rabbit ventricular muscle monitored with calcium-selective microelectrodes.

Transient changes of extracellular free calcium in rabbit ventricular muscle under nonsteady state conditions were measured with double-barreled calcium microelectrodes. Resumption of stimulation after a rest interval produces a cumulative decrease of extracellular free calcium often by more than 10% (with bulk extracellular free calcium = 0.2 mM). The extracellular free calcium returns to the bulk value as a new steady state is achieved. The changes of extracellular free calcium recorded presumably represent net calcium uptake and loss by cardiac muscle cells. These cumulative extracellular free calcium depletions are blocked by 0.5 mM cobalt and 1 microM nifedipine and are increased to 167 +/- 11% of control by the calcium agonist Bay k 8644 (1 microM) and to 620 +/- 150% of control by increasing stimulus frequency from 0.2-2 Hz. Caffeine (10 mM) inhibits the cumulative extracellular free calcium depletions, probably by rendering the sarcoplasmic reticulum unable to accumulate calcium. It is proposed that the extracellular free calcium depletions recorded represent, in large part, calcium which has entered the cells and has been taken up by the sarcoplasmic reticulum (which had become depleted of calcium during the rest interval). Nifedipine and cobalt inhibit these cumulative depletions presumably by preventing the calcium entry which could subsequently be accumulated by the sarcoplasmic reticulum. The net cellular calcium uptake produced by such a post-rest stimulation protocol can also be inhibited by 1-3 microM acetylstrophanthidin and reduction of extracellular sodium to 70 mM. Acetylstrophanthidin and low extracellular sodium would be expected to shift the sodium-calcium exchange in favor of increased calcium uptake, which may, in turn, prevent the loss of sarcoplasmic reticulum calcium during the rest interval. This would limit the amount of calcium which the sarcoplasmic reticulum could take up with subsequent activation. In contrast to the results with caffeine, ryanodine (1 microM) increases the magnitude and rate of calcium uptake after a rest interval, indicative of a fundamental difference in the actions of caffeine and ryanodine. When stimulation is stopped in the presence of ryanodine, extracellular free calcium increases much faster than in control. This suggests that ryanodine may enhance calcium uptake by the sarcoplasmic reticulum during repetitive stimulation and may enhance calcium efflux from the sarcoplasmic reticulum during quiescence. These experiments provide insight into transsarcolemmal calcium movements and certain aspects of cellular calcium regulation.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Asymmetry of neuronal activity during extracellular microelectrode recording from left and right human temporal lobe neocortex during rhyming and line-matching.

Recordings of neuronal activity in humans have identified few correlates of the known hemispheric asymmetries of functional lateralization. Here, we examine single-unit activity recorded from both hemispheres during two delayed match-to-sample tasks that show strong hemispheric lateralization based on lesion effects; a line-matching (LM) task related to the right hemisphere, and a rhyming (RHY) task related to the left. Nineteen neuronal populations were recorded with extracellular microelectrodes from the left temporal neocortex of 11 awake patients, and 18 from the right in 9 patients during anterior temporal lobectomy for complex partial seizures under local anesthesia. All subjects were left hemisphere dominant for language. Twelve (32%) populations exhibited statistically significant changes in activity at p <.05. Although changes in firing frequency were recorded from both hemispheres during both tasks, the RHY task elicited changes in activity several hundred milliseconds earlier on the left side than on the right. The LM task, on the other hand, induced changes earlier on the right side than on the left. Both hemispheres contained units active during verbal responses regardless of which behavior elicited the response. Our results indicate that cerebral dominance is reflected in earlier neuronal activity in the anterior temporal lobe during tasks lateralized to that hemisphere.

Adult↗

Sympathetic response to oral carbohydrate administration. Evidence from microelectrode nerve recordings.

Microneurography was used to measure sympathetic outflow in human muscle nerves (MSA) for up to 90 min after the ingestion of 100 g D-glucose, 75.8 g D-xylose, intravenous D-glucose (0.35 g/kg), and 300 ml water. 19 healthy subjects were examined using a microelectrode positioned in the right peroneal nerve. MSA increased from 21 +/- 0.9 bursts/min at rest to 36.9 +/- 4.3 bursts/min 30 min after ingestion of D-glucose and from 18.9 +/- 2.9 to 26.3 +/- 3.4 bursts/min 30 min after D-xylose. The increase in MSA was already significant by 15 min. MSA had not returned to the basal level after 90 min. Neither intravenous D-glucose nor water intake enhanced MSA. MSA increased in parallel with plasma norepinephrine, and a significant correlation (r = 0.55; P less than 0.001) was observed between the plasma insulin concentration and MSA after D-glucose ingestion. In three subjects the outflow of sympathetic nerve activity to the skin was examined after oral D-glucose and no change was observed, emphasizing the differentiated nature of the sympathetic nerve response to carbohydrate. Multiple factors such as insulin alone, hemodynamic adjustment to splanchnic vasodilation, and gastrointestinal distension are probably involved in the increased muscle nerve sympathetic outflow after carbohydrate ingestion.

Adult↗

Serum factor blocks neuromuscular transmission in myasthenia gravis: electrophysiologic study with intracellular microelectrodes.

We studied the effects of normal and myasthenic sera on the miniature endplate potential (MEPP) and resting membrane potential (RP) of rat muscle in vitro by conventional intracellular microelectrode techniques. Normal sera had little or no effect on either the amplitude or frequency of MEPP or RP. On the other hand, MEPP amplitude was reduced in each of nine muscles during exposure to myasthenic sera; five of these muscles showed a significant difference, by student's t-test, from the values in a control solution. The half decay time of diminished MEPP remained unchanged. MEPP frequency and RP were not affected by myasthenic sera. The reduced amplitude of the MEPP was almost completely restored when the muscle was washed with a control solution for more than 30 minutes. These observations indicate that myasthenic sera contain factors that bind reversibly with the acetylcholine receptor and reduce postsynaptic responses to acetylcholine.

Adult↗

Analysis of Na+, Cl-, K+, H+ and NH4+ concentration gradients adjacent to the surface of anal papillae of the mosquito Aedes aegypti: application of self-referencing ion-selective microelectrodes.

Ion concentration gradients adjacent to the surface of the anal papillae of larvae of the mosquito Aedes aegypti were measured using self-referencing ion-selective microelectrodes. The gradients were used to calculate estimates of ion fluxes into and out of the papillae. There was a net influx of Na+, Cl- and K+ from the bathing medium and a net efflux of acid and NH4+. No Ca2+ gradients were detectable. Na+ and Cl- influx occurred against a concentration gradient suggesting active transport. Although Na+, Cl- and NH4+ gradients were uniform along the length of the papillae, the proximal regions of the papillae in vivo revealed significantly higher H+ and K+ gradients compared with distal regions. The calculated ion fluxes at the papillae are sufficient for complete Na+, K+ and Cl- haemolymph replacement in approximately 4 h with external ion concentrations of 5 mmol l(-1). Ion gradients were also detected adjacent to the surface of isolated papillae; however, Na+ and H+ gradients were higher, and Cl- gradients were lower relative to papillae in vivo. The results support previous findings that the anal papillae of mosquito larvae are important structures for ion regulation, and suggest that these structures may be used for the excretion of nitrogenous waste.

Aedes↗

In situ analysis of pH gradients in mosquito larvae using non-invasive, self-referencing, pH-sensitive microelectrodes.

The alkaline environment, pH approximately 11, in the anterior midgut lumen of mosquito larvae is essential for normal nutrition and development. The mechanism of alkalization is, however, unknown. Although evidence from immunohistochemistry, electron microscopy and electrophysiology suggests that a V-ATPase is present in the basal membranes of the epithelial cells, its physiological role in the alkalization process has not been demonstrated. To investigate a possible role of the V-ATPase in lumen alkalization, pH gradients emanating from the hemolymph side of the midgut in semi-intact mosquito larvae were measured using non-invasive, self-referencing, ion-selective microelectrodes (SERIS). Large H+ concentration gradients, with highest concentrations close to the basal membrane (outward [H+] gradients), were found in the anterior midgut, whereas much smaller gradients, with concentrations lowest close to this membrane (inward [H+] gradients), were found in the gastric caeca and posterior midgut. Similar region-specific pH gradients, with consistent anterior-to-posterior profiles, were observed in individuals of two Aedes species, Aedes aegypti from semi-tropical Florida and Aedes canadensis from north-temperate Massachusetts. The gradients remained in a steady state for up to 6 h, the maximum duration of the recordings. Bafilomycin A1 (10(-5), 10(-7 )mol x l(-1)) on the hemolymph side greatly diminished the [H+] gradients in the anterior midgut but had no effect on the gradients in the gastric caecum and posterior midgut. These physiological data are consistent with the previous findings noted above. Together, they support the hypothesis that a basal, electrogenic H+ V-ATPase energizes luminal alkalization in the anterior midgut of larval mosquitoes.

Aedes↗