: The levels of several neurochemicals, i.e., uric acid (UA), dopamine (DA), dihydroxyphenylaceti... more : The levels of several neurochemicals, i.e., uric acid (UA), dopamine (DA), dihydroxyphenylacetic acid, and 5‐hydroxyindoleacetic acid, collected daily from the rat striatum with either fixed or removable microdialysis probes for 7 days after surgery were compared. The implantation of the fixed cannula was followed by a 10‐fold increase in the UA content in the dialysates collected from the first day after surgery onward and by a steady decrease in dihydroxyphenylacetic acid levels, whereas those of DA remained fairly stable. With the removable cannula system, only a smaller, transient increase in UA during the first 3 days after surgery was observed, with no change in DA or monoamine metabolites. The glial reaction around the cannula tracks was assessed by both quantitative histological techniques and measuring the glutamine levels in the dialysates collected at the time of surgery and 7 days later. Both the glial cell number and nuclear size, as well as the glutamine outflow, were considerably larger in the animals implanted with the fixed probes. It is, therefore, likely that the UA levels in the dialysate reflect the glial reaction to the probe. The suitability of the removable probe system for behavioral experiments involving repeated microdialysis sampling was illustrated in an experiment showing that the DA release in the nucleus accumbens of male rats assessed daily at postsurgery days 5–10 was virtually identical in three alternating sessions of sexual behavior as was the smaller release of this neurotransmitter detected during intervening nonsexual social interactions.
A wealth of pharmacological studies suggest that nitric oxide (NO) generated in the corpus cavern... more A wealth of pharmacological studies suggest that nitric oxide (NO) generated in the corpus cavernosum is a main molecular mediator of penile erection. However, the physiological levels of NO in the corpora and their possible changes during penile erection have remained unknown for want of suitable methodologies. We have adapted a voltammetric procedure, derived from Malinski's method, for assessing NO levels in the penis in vivo. Differential normal pulse voltammetry with carbon fiber electrodes (30 microm) coated with a polymeric porphyrin and Nafion was used to measure the NO oxidation current in the corpora cavernosa of urethane-anesthetized rats. The intracavernous pressure was monitored simultaneously. A NO oxidation peak was consistently detected at approximately 650 mV both in NO solutions and in the corpora in vivo. The changes in the NO signals observed in vitro were consistent with the concentration values measured by chemiluminescence. The NO signal recorded in vivo increased following cavernous nerve stimulation and was greatly decreased by intracavernous injections of several inhibitors of the neuronal and endothelial NO synthase isoenzymes. Such results agree with our previous studies using this methodology and substantiate further its validity for monitoring the physiological changes in NO levels in the penis.
A microcomputer-assisted curve-fitting procedure was developed for the quantitative estimation of... more A microcomputer-assisted curve-fitting procedure was developed for the quantitative estimation of the components of the mixed "catechol peak" recorded with differential normal pulse voltammetry (DNPV) at electrochemically pretreated carbon fiber microelectrodes in the living brain. The contribution of each of the relevant electroactive species is fitted by a normal probability function, the parameters of which are previously determined in vitro for each electrode and substance. The voltammogram is thus modeled as a mixture of normal curves corresponding to the individual oxidizable substances plus a low order polynomial approximating the baseline. In a former approach the function was solved by linear least squares techniques. As a further improvement, we now propose a non-linear model of the voltammogram and a Gauss-Newton iterative algorithm with stepwise regression for parameter estimation. This report shows the application of the method for the resolution of the dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) components of the DNPV signal recorded from the striatum of freely moving animals in response to amphetamine and pargyline. The method was validated by the chemical assay of contralateral microdialysates collected simultaneously. The changes detected by both methodologies were closely parallel, with highly significant correlation coefficients (0.87 and 0.99 for DA and DOPAC, respectively, P less than 0.001). This study further illustrates that the in vivo voltammetry methodology can be improved substantially by incorporating a suitable mathematical treatment of the electrochemical signals.
Clinical intervention in neurological disorders is almost invariably achieved using chemical agen... more Clinical intervention in neurological disorders is almost invariably achieved using chemical agents that act on neuromediator-related sites, suggesting that intercellular chemical signaling plays a major role in determining the properties of neural networks. A variety of microvoltammetric sensors and techniques have been developed over the last 25 years to study neuromediators in intact brain in vivo, and in isolated tissues, for animal models of behavior and disease. This review, with over 600 citations, considers the advantages and limitations of the different approaches, including progress in biosensor design, illustrated by studies on the neurochemical bases of a wide variety of behaviors.
: The levels of several neurochemicals, i.e., uric acid (UA), dopamine (DA), dihydroxyphenylaceti... more : The levels of several neurochemicals, i.e., uric acid (UA), dopamine (DA), dihydroxyphenylacetic acid, and 5‐hydroxyindoleacetic acid, collected daily from the rat striatum with either fixed or removable microdialysis probes for 7 days after surgery were compared. The implantation of the fixed cannula was followed by a 10‐fold increase in the UA content in the dialysates collected from the first day after surgery onward and by a steady decrease in dihydroxyphenylacetic acid levels, whereas those of DA remained fairly stable. With the removable cannula system, only a smaller, transient increase in UA during the first 3 days after surgery was observed, with no change in DA or monoamine metabolites. The glial reaction around the cannula tracks was assessed by both quantitative histological techniques and measuring the glutamine levels in the dialysates collected at the time of surgery and 7 days later. Both the glial cell number and nuclear size, as well as the glutamine outflow, were considerably larger in the animals implanted with the fixed probes. It is, therefore, likely that the UA levels in the dialysate reflect the glial reaction to the probe. The suitability of the removable probe system for behavioral experiments involving repeated microdialysis sampling was illustrated in an experiment showing that the DA release in the nucleus accumbens of male rats assessed daily at postsurgery days 5–10 was virtually identical in three alternating sessions of sexual behavior as was the smaller release of this neurotransmitter detected during intervening nonsexual social interactions.
A wealth of pharmacological studies suggest that nitric oxide (NO) generated in the corpus cavern... more A wealth of pharmacological studies suggest that nitric oxide (NO) generated in the corpus cavernosum is a main molecular mediator of penile erection. However, the physiological levels of NO in the corpora and their possible changes during penile erection have remained unknown for want of suitable methodologies. We have adapted a voltammetric procedure, derived from Malinski's method, for assessing NO levels in the penis in vivo. Differential normal pulse voltammetry with carbon fiber electrodes (30 microm) coated with a polymeric porphyrin and Nafion was used to measure the NO oxidation current in the corpora cavernosa of urethane-anesthetized rats. The intracavernous pressure was monitored simultaneously. A NO oxidation peak was consistently detected at approximately 650 mV both in NO solutions and in the corpora in vivo. The changes in the NO signals observed in vitro were consistent with the concentration values measured by chemiluminescence. The NO signal recorded in vivo increased following cavernous nerve stimulation and was greatly decreased by intracavernous injections of several inhibitors of the neuronal and endothelial NO synthase isoenzymes. Such results agree with our previous studies using this methodology and substantiate further its validity for monitoring the physiological changes in NO levels in the penis.
A microcomputer-assisted curve-fitting procedure was developed for the quantitative estimation of... more A microcomputer-assisted curve-fitting procedure was developed for the quantitative estimation of the components of the mixed "catechol peak" recorded with differential normal pulse voltammetry (DNPV) at electrochemically pretreated carbon fiber microelectrodes in the living brain. The contribution of each of the relevant electroactive species is fitted by a normal probability function, the parameters of which are previously determined in vitro for each electrode and substance. The voltammogram is thus modeled as a mixture of normal curves corresponding to the individual oxidizable substances plus a low order polynomial approximating the baseline. In a former approach the function was solved by linear least squares techniques. As a further improvement, we now propose a non-linear model of the voltammogram and a Gauss-Newton iterative algorithm with stepwise regression for parameter estimation. This report shows the application of the method for the resolution of the dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) components of the DNPV signal recorded from the striatum of freely moving animals in response to amphetamine and pargyline. The method was validated by the chemical assay of contralateral microdialysates collected simultaneously. The changes detected by both methodologies were closely parallel, with highly significant correlation coefficients (0.87 and 0.99 for DA and DOPAC, respectively, P less than 0.001). This study further illustrates that the in vivo voltammetry methodology can be improved substantially by incorporating a suitable mathematical treatment of the electrochemical signals.
Clinical intervention in neurological disorders is almost invariably achieved using chemical agen... more Clinical intervention in neurological disorders is almost invariably achieved using chemical agents that act on neuromediator-related sites, suggesting that intercellular chemical signaling plays a major role in determining the properties of neural networks. A variety of microvoltammetric sensors and techniques have been developed over the last 25 years to study neuromediators in intact brain in vivo, and in isolated tissues, for animal models of behavior and disease. This review, with over 600 citations, considers the advantages and limitations of the different approaches, including progress in biosensor design, illustrated by studies on the neurochemical bases of a wide variety of behaviors.
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Papers by Manuel Mas