RESUMO
The mechanisms of action of the rapid antidepressant effects of ketamine, an N-methyl-D-aspartate glutamate receptor antagonist, have not been fully elucidated. This study examined the effects of ketamine on ligand binding to a metabotropic glutamatergic receptor (mGluR5) in individuals with major depressive disorder (MDD) and healthy controls. Thirteen healthy and 13 MDD nonsmokers participated in two [11C]ABP688 positron emission tomography (PET) scans on the same day-before and during intravenous ketamine administration-and a third scan 1 day later. At baseline, significantly lower [11C]ABP688 binding was detected in the MDD as compared with the control group. We observed a significant ketamine-induced reduction in mGluR5 availability (that is, [11C]ABP688 binding) in both MDD and control subjects (average of 14±9% and 19±22%, respectively; P<0.01 for both), which persisted 24 h later. There were no differences in ketamine-induced changes between MDD and control groups at either time point (P=0.8). A significant reduction in depressive symptoms was observed following ketamine administration in the MDD group (P<0.001), which was associated with the change in binding (P<0.04) immediately after ketamine. We hypothesize that glutamate released after ketamine administration moderates mGluR5 availability; this change appears to be related to antidepressant efficacy. The sustained decrease in binding may reflect prolonged mGluR5 internalization in response to the glutamate surge.
Assuntos
Depressão/diagnóstico por imagem , Ketamina/metabolismo , Receptor de Glutamato Metabotrópico 5/efeitos dos fármacos , Adulto , Antidepressivos/farmacologia , Encéfalo/metabolismo , Radioisótopos de Carbono , Estudos de Casos e Controles , Transtorno Depressivo Maior/diagnóstico por imagem , Transtorno Depressivo Maior/metabolismo , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Ácido Glutâmico/metabolismo , Humanos , Ketamina/farmacologia , Masculino , Tomografia por Emissão de Pósitrons/métodos , Receptor de Glutamato Metabotrópico 5/metabolismoRESUMO
Sprague Dawley rats (10/sex/group) were given a single intravenous (iv) dose of CUMI-101 to determine acute toxicity of CUMI-101 and radiation dosimetry estimations were conducted in baboons with [(11)C]CUMI-101. Intravenous administration of CUMI-101 did not produce overt biologically or toxicologically significant adverse effects except transient hypoactivity immediately after dose in the mid- and high-dose groups, which is not considered to be a dose-limiting toxic effect. No adverse effects were observed in the low-dose group. The no observed adverse effect level (NOAEL) is considered to be 44.05 µg/kg for a single iv dose administration in rats. The maximum tolerated dose (MTD) was estimated to be 881 µg/kg for a single iv dose administration. The Medical Internal Radiation Dose (MIRDOSE) estimates indicate the maximum permissible single-study dosage of [(11)C]CUMI-101 in humans is 52 mCi with testes and urinary bladder as the critical organ for males and females, respectively.
Assuntos
Piperazinas/farmacocinética , Agonistas do Receptor 5-HT1 de Serotonina/farmacocinética , Triazinas/farmacocinética , Animais , Radioisótopos de Carbono , Feminino , Ligantes , Masculino , Papio , Piperazinas/toxicidade , Tomografia por Emissão de Pósitrons , Radiometria , Ratos , Ratos Sprague-Dawley , Agonistas do Receptor 5-HT1 de Serotonina/toxicidade , Distribuição Tecidual , Triazinas/toxicidadeRESUMO
We have studied the effect of ascorbic acid on voltage-dependent calcium channels in pancreatic beta cells. Using the whole-cell and perforated-patch variants of the patch clamp technique to record calcium tail currents, we have shown that the slowly deactivating (SD) calcium channel, which is similar to the T-type channel in other cells, is inhibited in a voltage-dependent manner by ascorbic acid (AA). The other channels that carry inward current in beta cells, FD calcium channels and sodium channels, are unaffected by AA. Ascorbic acid causes a voltage-dependent decrease in the magnitude of the SD channel conductance which can be explained by the hypothesis that approximately 50-60% of the channels have their voltage dependence shifted by approximately 62 mV in the depolarizing direction. Thus, ascorbate appears to modify only a fraction of the SD channels. The activation kinetics of the ascorbate-modified channels are slower than control channels in a manner that is consistent with this hypothesis. Deactivation and inactivation kinetics are unaffected by ascorbate. These effects of ascorbate require metal ions, and it appears that some of the activity of ascorbate is due to a product of its metal catalyzed oxidation, perhaps dehydroascorbate.
Assuntos
Ácido Ascórbico/farmacologia , Canais de Cálcio/metabolismo , Ilhotas Pancreáticas/metabolismo , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio/efeitos dos fármacos , Células Cultivadas , Eletrodos , Eletrofisiologia , Insulina/metabolismo , Ilhotas Pancreáticas/efeitos dos fármacos , Cinética , Masculino , Metais/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
The metabotropic glutamate receptor subtype 5 (mGluR5) has been implicated in the pathophysiology of mood and anxiety disorders and is a potential treatment target in major depressive disorder (MDD). This study compared brain mGluR5 binding in elderly patients suffering from MDD with that in elderly healthy volunteers using positron emission tomography (PET) and [(11)C]ABP688. Twenty elderly (mean age: 63.0 ± 6.3) subjects with MDD and twenty-two healthy volunteers in the same age range (mean age: 66.4 ± 7.3) were examined with PET after a single bolus injection of [(11)C]ABP688, with many receiving arterial sampling. PET images were analyzed on a region of interest and a voxel level to compare mGluR5 binding in the brain between the two groups. Differences in [(11)C]ABP688 binding between patients with early- and late-onset depression were also assessed. In contrast to a previously published report in a younger cohort, no significant difference in [(11)C]ABP688 binding was observed between elderly subjects with MDD and healthy volunteers. [(11)C]ABP688 binding was also similar between subgroups with early- or late-onset depression. We believe this is the first study to examine mGluR5 expression in depression in the elderly. Although future work is required, results suggest potential differences in the pathophysiology of elderly depression versus depression earlier in life.
Assuntos
Encéfalo/metabolismo , Radioisótopos de Carbono , Transtorno Depressivo Maior/metabolismo , Oximas , Tomografia por Emissão de Pósitrons , Piridinas , Receptor de Glutamato Metabotrópico 5/metabolismo , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos PilotoRESUMO
BACKGROUND: Reduced dopaminergic transmission has been implicated in the pathophysiology of major depression. The aim of the present study was to measure striatal D(2) receptor availability and amphetamine-induced dopamine release in nonpsychotic, unmedicated, unipolar patients during an episode of major depression. METHODS: The striatal equilibrium specific to nonspecific partition coefficient (V(3)") of the D(2) receptor antagonist [(123)I]IBZM was measured with single photon emission computerized tomography before and after amphetamine administration in 9 depressed subjects and 10 matched healthy control subjects. RESULTS: No significant differences were observed in preamphetamine D(2) receptor availability between depressed patients (0.73 +/- 0.08) and control subjects (0.78 +/- 0.10, p =.23). Amphetamine-induced reduction in [(123)I]IBZM V(3)" (DeltaV(3)") was similar in depressed patients (-9.8 +/- 5.5%) and control subjects (-7.8 +/- 2.5%, p =.32). Amphetamine induced a transient improvement in symptomatology in depressed patients, but this improvement did not correlate with [(123)I]IBZM DeltaV(3)". CONCLUSIONS: This study did not replicate previously reported alterations in striatal D(2) receptor density in depressed patients and suggests that stimulant-induced dopamine release is not altered in major depression.
Assuntos
Anfetamina/farmacologia , Corpo Estriado/fisiopatologia , Transtorno Depressivo Maior/fisiopatologia , Dopamina/metabolismo , Receptores de Dopamina D2/fisiologia , Tomografia Computadorizada de Emissão de Fóton Único , Adolescente , Adulto , Anfetamina/farmacocinética , Benzamidas , Corpo Estriado/diagnóstico por imagem , Transtorno Depressivo Maior/diagnóstico por imagem , Transtorno Depressivo Maior/psicologia , Feminino , Humanos , Radioisótopos do Iodo , Masculino , Pessoa de Meia-Idade , Pirrolidinas , Receptores de Dopamina D2/efeitos dos fármacos , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
Serotonin 5-HT(1A) receptors are implicated in the pathophysiology of neuropsychiatric conditions. The goal of this study was to evaluate methods to derive 5-HT(1A) receptor parameters in the human brain with positron emission tomography (PET) and [carbonyl-(11)C]WAY 100635. Five healthy volunteer subjects were studied twice. Three methods of analysis were used to derive the binding potential (BP), and the specific to nonspecific equilibrium partition coefficient (k3/k4). Two methods, kinetic analysis based on a three compartment model and graphical analysis, used the arterial plasma time-activity curves as the input function to derive BP and k3/k4. A third method, the simplified reference tissue model (SRTM), derived the input function from uptake data of a region of reference, the cerebellum, and provided only k3/k4. All methods provided estimates of regional 5-HT(1A) receptor parameters that were highly correlated. Results were consistent with the known distribution of 5-HT(1A) receptors in the human brain. Compared with kinetic BP, graphical analysis slightly underestimated BP, and this phenomenon was mostly apparent in small size-high noise regions. Compared with kinetic k3/k4, the reference tissue method underestimated k3/k4 and the underestimation was apparent primarily in regions with high receptor density. Derivation of BP by both kinetic and graphical analysis was highly reliable, with an intraclass correlation coefficient (ICC) of 0.84 +/- 0.14 (mean +/- SD of 15 regions) and 0.84 +/- 0.19, respectively. In contrast, the reliability of k3/k4 was lower, with ICC of 0.53 +/- 0.28, 0.47 +/- 0.28, and 0.55 +/- 0.29 for kinetic, graphical, and reference tissue methods, respectively. In conclusion, derivation of BP by kinetic analysis using the arterial plasma input function appeared as the method of choice because of its higher test-retest reproducibility, lower vulnerability to experimental noise, and absence of bias.
Assuntos
Encéfalo/metabolismo , Receptores de Serotonina/metabolismo , Adulto , Encéfalo/diagnóstico por imagem , Cerebelo/metabolismo , Estudos de Avaliação como Assunto , Humanos , Cinética , Masculino , Modelos Biológicos , Piperazinas/farmacocinética , Piridinas/farmacocinética , Receptores de Serotonina/sangue , Receptores 5-HT1 de Serotonina , Reprodutibilidade dos Testes , Antagonistas da Serotonina/farmacocinética , Tomografia Computadorizada de EmissãoRESUMO
Augmentation of selective serotonin reuptake inhibitors (SSRIs) therapy by the 5-HT(1A) receptor agent pindolol may reduce the delay between initiation of antidepressant treatment and clinical response. This hypothesis is based on the ability of pindolol to block 5-HT(1A) autoreceptors in the dorsal raphe nuclei (DRN) and to potentiate the increase in 5-HT transmission induced by SSRIs. However, placebo-controlled clinical studies of pindolol augmentation of antidepressant therapy have reported inconsistent results. Here, we evaluated the occupancy of 5-HT(1A) receptors during treatment with pindolol controlled release (CR) in nine healthy volunteers with Positron Emission Tomography and [11C]WAY 100635. Subjects were studied four times: at baseline, following one week of pindolol CR 7.5 mg/day (4 and 10 hrs post dose), and following one dose of pindolol CR 30 mg(4 hrs post dose). Occupancy of the DRN was 40 +/- 29% on scan 2, 38 +/- 26% on scan 3, and 64 +/- 15% on scan 4. The average occupancy in all other regions was significantly lower at each doses (18 +/- 5% on scan 2, 12 +/- 3% on scan 3, and 42 +/- 4% on scan 4). These results suggest that the blockade in the DRN reached in clinical studies (7.5 mg/day) might be too low and variable to consistently augment the therapeutic effect of SSRIs. However, these data indicate that pindolol exhibits in vivo selectivity for the DRN 5-HT(1A) autoreceptors. As DRN selectivity is desirable for potentiation of 5-HT function, this observation represents an important proof of concept for the development of 5-HT(1A) agents in this application.
Assuntos
Transtornos do Humor/tratamento farmacológico , Pindolol/farmacologia , Piperazinas/uso terapêutico , Piridinas/uso terapêutico , Núcleos da Rafe/metabolismo , Receptores de Serotonina/efeitos dos fármacos , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Adulto , Antidepressivos/uso terapêutico , Encéfalo/efeitos dos fármacos , Humanos , Cinética , Imageamento por Ressonância Magnética , Masculino , Pindolol/administração & dosagem , Pindolol/sangue , Pindolol/uso terapêutico , Piperazinas/sangue , Piperazinas/farmacocinética , Piridinas/sangue , Piridinas/farmacocinética , Receptores de Neurotransmissores/fisiologia , Receptores 5-HT1 de Serotonina , Inibidores Seletivos de Recaptação de Serotonina/uso terapêutico , Transmissão Sináptica/fisiologia , Tomografia Computadorizada de EmissãoRESUMO
UNLABELLED: Abnormal brain regional densities of serotonin (5-hydroxytryptamine [5-HT]) transporters have been reported in postmortem studies in several neuropsychiatric conditions, such as major depression and schizophrenia. trans-1,2,3,5,6,10-beta-Hexahydro-6-[4-(methylthio)phenyl]pyrrolo-[2,1-a]-isoquinoline ([11C]McN 5652) is the first PET radioligand successfully developed to label 5-HT transporters in the living human brain. The purpose of this study was to develop an imaging protocol and analytic method to measure regional 5-HT transporter binding potential (BP) with [11C]McN 5652 in humans. METHODS: The arterial input function and brain uptake of (+)-[11C]McN 5652 and (-)-[11C]McN 5652, the active and inactive enantiomers, respectively, were measured in 6 healthy volunteers. RESULTS: (+)-[11C]McN 5652 concentrated in brain regions rich in 5-HT transporters (midbrain, thalamus, basal ganglia, and medial temporal lobe structures), whereas the uptake of (-)-[11C]McN 5652 was more uniformly distributed. Total distribution volumes (V(T)) were derived using kinetic 2-compartment analysis and graphic analysis. V(T) derived by both methods were highly correlated. (+)-[11C]McN 5652 regional V(T) ranged from 18 +/- 2 mL/g in the cerebellum to 46 +/- 13 mL/g in the midbrain. (-)-[11C]McN 5652 regional VT ranged from 10 +/- 2 mL/g in the cerebellum to 14 +/- 3 mL/g in the thalamus. (+)-[11C]McN 5652 V(T) were higher than (-)-[11C]McN 5652 V(T) in all regions, including the cerebellum, a region devoid of 5-HT transporters. Blocking experiments were also performed in baboons with saturating doses of citalopram and in humans with nonsaturating doses of paroxetine. Cerebellar and neocortical (+)-[11C]McN 5652 V(T) were unaffected by pretreatment with 5-HT transporter blockers. In areas of high receptor concentration (midbrain, caudate, and thalamus) 5-HT transporter blockers decreased (+)-[11C]McN 5652 V(T) to the level of cerebellum (+)-[11C]McN 5652 V(T). CONCLUSION: These experiments indicate that the use of the difference between (+)- and (-)-[11C]McN 5652 V(T) to define specific binding to 5-HT transporters leads to an overestimation of specific binding. 5-HT transporter BP was derived as the difference between the regional and cerebellar (+)-[11C]McN 5652 V(T). BP values were in good agreement with the distribution of 5-HT transporters in the human brain, except for regions of relatively low 5-HT transporter concentration, such as the prefrontal cortex, where no specific binding was detected using (+)-[11C]McN 5652. (+)-[11C]McN 5652 is an appropriate radiotracer to quantify 5-HT transporters in regions with relatively high concentration of 5-HT transporters, such as the midbrain, thalamus, and basal ganglia, and should prove useful in elucidating abnormalities of 5-HT transmission in neuropsychiatric conditions.
Assuntos
Encéfalo/metabolismo , Proteínas de Transporte/metabolismo , Radioisótopos do Iodo/farmacocinética , Isoquinolinas/farmacocinética , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana Transportadoras , Proteínas do Tecido Nervoso , Antagonistas da Serotonina/farmacocinética , Adulto , Encéfalo/diagnóstico por imagem , Proteínas de Transporte/análise , Humanos , Cinética , Imageamento por Ressonância Magnética , Masculino , Glicoproteínas de Membrana/análise , Mesencéfalo/diagnóstico por imagem , Mesencéfalo/metabolismo , Valores de Referência , Proteínas da Membrana Plasmática de Transporte de Serotonina , Distribuição Tecidual , Tomografia Computadorizada de EmissãoRESUMO
1. Alzheimer's Disease (AD) is accompanied by a disruption in iron metabolism. There is no universally accepted method for detecting brain iron. 2. The authors have developed a novel "ratio" method which uses the red nucleus as an internal reference. We postulated that this method would improve our sensitivity in detecting differences in MRI signal intensities and that it would allow us to measure brain iron deposition. 3. The ratio method reasonably reproduced previous reports of the normal deposition of iron in brain that occurs with aging. It failed to distinguish any differences in three brain areas: putamen, left frontal lobe and whole slice in AD patients versus age and sex matched controls. 4. It also failed to detect differences with AD progression or severity. 5. The ratio method itself warrants further investigation.
Assuntos
Envelhecimento/patologia , Doença de Alzheimer/patologia , Imageamento por Ressonância Magnética/métodos , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/metabolismo , Doença de Alzheimer/metabolismo , Química Encefálica , Humanos , Processamento de Imagem Assistida por Computador , Ferro/metabolismo , Pessoa de Meia-Idade , Putamen/metabolismo , Putamen/patologiaRESUMO
In several positron-emission tomography studies of human subjects, analyses of data from the 5-hydroxytryptamine(1A) (5-HT(1A)) receptor radioligand, [(11)C]WAY-100635 ¿[carbonyl-(11)C]N-(2-(4-methoxy-phenyl)-1-piperazin-1-yl)ethyl)-N -(2 -pyridyl)cyclohexanecarboxamide¿ have shown a discrepancy between the outcome measure k(3)/k(4) (binding potential normalized to cerebellum) as estimated by the simplified reference region method and results obtained by conventional kinetic modeling with an arterial input function. The reference region method has yielded results that are lower than the conventional approach, with the relative underestimation appearing to be an increasing function of k(3)/k(4). We performed simulations on idealized data to identify the source of the discrepancy. Both the simplified reference tissue model (SRTM) and the original full reference tissue model (FRTM) were tested to determine (a) if the error in estimated k(3)/k(4) is dependent on the blood flow in the region of interest relative to the blood flow in the region of reference (R(1)) and on the receptor density in the region of interest (true k(3)/k(4)), and (b) which violation of the reference model assumptions were responsible for this effect. FRTM returned parameter estimates that were independent and accurate if the reference region was constructed precisely as a one-tissue compartment model. SRTM overestimated k(3)/k(4) when the reference region was constructed as a one-tissue compartment model and underestimated k(3)/k(4) when the reference region was constructed as a two-tissue compartment model (which is the case for [(11)C]WAY-100635). In both cases, the magnitude of the error in k(3)/k(4) returned by SRTM was dependent on true R(1) and true k(3)/k(4). In conclusion, the SRTM is associated with a bias in the derivation of k(3)/k(4) that is not a simple scaling factor. This magnitude of these errors should be carefully evaluated for each new radioligand.
Assuntos
Radioisótopos de Carbono , Piperazinas/farmacocinética , Piridinas/farmacocinética , Receptores de Serotonina/análise , Antagonistas da Serotonina/farmacocinética , Humanos , Modelos Biológicos , Racloprida/farmacocinética , Receptores 5-HT1 de SerotoninaRESUMO
Preclinical studies in rodents suggest that augmentation of serotonin reuptake inhibitors (SSRIs) therapy by the 5-hydroxytryptamine(1A) (5-HT(1A)) receptor agent pindolol might reduce the delay between initiation of treatment and antidepressant response. This hypothesis is based on the ability of pindolol to potentiate the increase in serotonin (5-HT) transmission induced by SSRIs, an effect achieved by blockade of the 5-HT(1A) autoreceptors in the dorsal raphe nuclei (DRN). However, placebo-controlled clinical studies of pindolol augmentation of antidepressant therapy have reported inconsistent results. Here, we evaluated the occupancy of 5-HT(1A) receptors following treatment with controlled release pindolol in nine healthy volunteers with positron-emission tomography (PET). Each subject was studied four times: at baseline (scan 1), following 1 week of oral administration of pindolol CR (7.5 mg/day) at peak level, 4 h after the dose (scan 2), and at 10 h following the dose (scan 3), and following one dose of pindolol CR (30 mg) (at peak level, 4 h) (scan 4). Pindolol occupancy of 5-HT(1A) receptors was evaluated in the DRN and cortical regions as the decrease in binding potential (BP) of the radiolabelled selective 5-HT(1A) antagonist [carbonyl-(11)C]WAY-100635 or [carbonyl-(11)C] N-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridyl)cyclohexa necarboxamide abbreviated as [(11)C]WAY-100635. Pindolol dose-dependently decreased [(11)C]WAY-100635 BP. Combining all the regions, occupancy was 20 +/- 8% at scan 2, 14 +/- 8% at scan 3, and 44 +/- 8% at scan 4. The results of this study suggest that at doses used in clinical studies of augmentation of the SSRI effect by pindolol (2.5 mg t.i.d.), the occupancy of 5-HT(1A) receptors is moderate and highly variable between subjects. This factor might explain the variable results obtained in clinical studies. On the other hand, at each dose tested, pindolol occupancy of 5-HT(1A) receptors was higher in the DRN compared to cortical regions, demonstrating a significant in vivo selectivity for DRN 5-HT(1A) autoreceptors relative to cortico-limbic postsynaptic receptors. This selectivity is necessary for the potentiation of 5-HT transmission, and this finding represents an important proof of concept in the development of 5-HT(1A) agents for this application. Early evaluation of new drugs with PET imaging will enable rapid screening of compounds based on DRN selectivity and more appropriate determination of doses for clinical trials.
Assuntos
Pindolol/metabolismo , Receptores de Serotonina/análise , Antagonistas da Serotonina/metabolismo , Tomografia Computadorizada de Emissão , Adulto , Química Encefálica , Humanos , Masculino , Pessoa de Meia-Idade , Núcleos da Rafe/química , Receptores 5-HT1 de SerotoninaRESUMO
Hypointensities (focal areas of decreased signal intensity) have been reported on T2 weighted magnetic resonance images (MRI) in normal aging and in some neurological disease processes. Increased concentrations of iron have been suggested as one cause of these hypointensities. In Alzheimer's Disease, data suggests that there is both a disruption in iron metabolism as well as the presence of T2 hypointensities. We endeavored to determine if the decreased signal intensities could be quantitatively determined and, if so, in what regions, in an effort to establish a non-invasive biological marker and diagnostic aide. We performed a quantitative analysis of the T2 signal intensities in 13 MRIs from AD patients and 16 age- and sex-matched control subjects. We found that while there were statistically significant differences in the intensities of the putamen and red nucleus, these differences were small. We were unable to detect differences in intensities in a whole slice or the frontal lobe. To our knowledge this is the first quantitative comparison of MRI signal intensities in Alzheimer's Disease.
Assuntos
Doença de Alzheimer/patologia , Encéfalo/patologia , Ferro/metabolismo , Idoso , Doença de Alzheimer/metabolismo , Encéfalo/metabolismo , Feminino , Lobo Frontal/metabolismo , Lobo Frontal/patologia , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Putamen/metabolismo , Putamen/patologiaRESUMO
We have used the nystatin perforated patch technique to study ionic currents in rat pancreatic beta-cells. The access resistance (Ra) between the pipette and the cell cytoplasm, measured by analyzing capacitive currents, decreased with a slow exponential time course (tau = 5.4 +/- 2.7 min) after seal formation. As Ra decreased, the magnitude of voltage-dependent K and Ca currents increased with a similar time course, and their activation kinetics became faster. After Ra stabilized, the macroscopic currents remained stable for up to an hour or more. When the final Ra was sufficiently low, Ca tail currents could be resolved which had properties similar to those recorded with the classical whole-cell technique. Two types of K channels could be characterized with perforated patch recordings of macroscopic K currents: (i) ATP-blockable K (KATP) channels which generate a time and voltage independent current that is blocked by glyburide and enhanced by pinacidil and (ii) voltage-dependent K (Kv) channels. Whole-cell recordings of KATP currents in the absence of ATP in the pipette showed that the maximum KATP conductance of the beta-cell was 83.8 +/- 40 nS. Perforated patch recordings show that the resting KATP conductance is 3.57 +/- 2.09 nS, which corresponds to about 4% of the channels being open in the intact beta-cell. In classical whole-cell recordings. Kv activation kinetics become faster during the first 10-15 min of recording, probably due to a dissipating Donnan potential. In perforated patch recordings where the Donnan potential is very small, Kv activation kinetics were nearly identical to the steady-state whole cell measurements.