Interaction Between Neurons and Glia in Aging and Disease, 2007
The functional activity in the brain is primarily composed of interplay between excitation and in... more The functional activity in the brain is primarily composed of interplay between excitation and inhibition. In any given region the output is based upon a complex processing of incoming signals that require both excitatory and inhibitory units. Moreover, these units must be regulated and balanced such that an integrated and finely tuned response is generated. In each of these units
Diseases of the brain account for much human suffering and place a huge burden on the health care... more Diseases of the brain account for much human suffering and place a huge burden on the health care systems. Thus, research into the pathology of brain diseases and improved pharmacotherapy is of significant value. In this respect, knowledge on malfunctions of metabolic homeostasis related to the neurotransmission process is still limited. As evident from this chapter, failure of the metabolic
Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism, 2014
Despite the well-established use of kainate as a model for seizure activity and temporal lobe epi... more Despite the well-established use of kainate as a model for seizure activity and temporal lobe epilepsy, most studies have been performed at doses giving rise to general limbic seizures and have mainly focused on neuronal function. Little is known about the effect of lower doses of kainate on cerebral metabolism and particularly that associated with astrocytes. We investigated astrocytic and neuronal metabolism in the cerebral cortex of adult mice after treatment with saline (controls), a subconvulsive or a mildly convulsive dose of kainate. A combination of [1,2-(13)C]acetate and [1-(13)C]glucose was injected and subsequent nuclear magnetic resonance spectroscopy of cortical extracts was employed to distinctively map astrocytic and neuronal metabolism. The subconvulsive dose of kainate led to an instantaneous increase in the cortical lactate content, a subsequent reduction in the amount of [4,5-(13)C]glutamine and an increase in the calculated astrocytic TCA cycle activity. In contr...
Diet supplementation with ketone bodies (acetoacetate and β-hydroxybuturate) or medium-length fat... more Diet supplementation with ketone bodies (acetoacetate and β-hydroxybuturate) or medium-length fatty acids generating ketone bodies has consistently been found to cause modest improvement of mental function in Alzheimer's patients. It was suggested that the therapeutic effect might be more pronounced if treatment was begun at a preclinical stage of the disease instead of well after its manifestation. The preclinical stage is characterized by decade-long glucose hypometabolism in brain, but ketone body metabolism is intact even initially after disease manifestation. One reason for the impaired glucose metabolism may be early destruction of the noradrenergic brain stem nucleus, locus coeruleus, which stimulates glucose metabolism, at least in astrocytes. These glial cells are essential in Alzheimer pathogenesis. The β-amyloid peptide Aβ interferes with their cholinergic innervation, which impairs synaptic function due to diminished astrocytic glutamate release. Aβ also reduces gluc...
Cultured astrocytes treated with siRNA to knock down glutamate dehydrogenase (GDH) were used to i... more Cultured astrocytes treated with siRNA to knock down glutamate dehydrogenase (GDH) were used to investigate whether this enzyme is important for the utilization of glutamate as an energy substrate. By incubation of these cells in media containing different concentrations of glutamate (range 100-500 µM) in the presence or in the absence of glucose, the metabolism of these substrates was studied by using tritiated glutamate or 2-deoxyglucose as tracers. In addition, the cellular contents of glutamate and ATP were determined. The astrocytes were able to maintain physiological levels of ATP regardless of the expression level of GDH and the incubation condition, indicating a high degree of flexibility with regard to regulatory mechanisms involved in maintaining an adequate energy level in the cells. Glutamate uptake was found to be increased in these cells when exposed to increasing levels of extracellular glutamate independently of the GDH expression level. Moreover, increased intracell...
The central nervous system (CNS) operates by a fine-tuned balance between excitatory and inhibito... more The central nervous system (CNS) operates by a fine-tuned balance between excitatory and inhibitory signalling. In this context, the inhibitory neurotransmission may be of particular interest as it has been suggested that such neuronal pathways may constitute 'command pathways' and the principle of 'dis-inhibition' leading ultimately to excitation may play a fundamental role (Roberts, E. (1974). Adv. Neurol., 5: 127-143). The neurotransmitter responsible for this signalling is gamma-aminobutyrate (GABA) which was first discovered in the CNS as a curious amino acid (Roberts, E., Frankel, S. (1950). J. Biol. Chem., 187: 55-63) and later proposed as an inhibitory neurotransmitter (Curtis, D.R., Watkins, J.C. (1960). J. Neurochem., 6: 117-141; Krnjevic, K., Schwartz, S. (1967). Exp. Brain Res., 3: 320-336). The present review will describe aspects of GABAergic neurotransmission related to homeostatic mechanisms such as biosynthesis, metabolism, release and inactivation. ...
Pyruvate recycling was studied in primary cultures of mouse cerebrocortical astrocytes, GABAergic... more Pyruvate recycling was studied in primary cultures of mouse cerebrocortical astrocytes, GABAergic cerebrocortical interneurons, and co-cultures consisting of both cell types by measuring production of [4-(13)C]glutamate from [3-(13)C]glutamate by aid of nuclear magnetic resonance spectroscopy. This change in the position of the label can only occur by entry of [3-(13)C]glutamate into the tricarboxylic acid (TCA) cycle, conversion of labeled alpha-ketoglutarate to malate or oxaloacetate, malic enzyme-mediated decarboxylation of malate to pyruvate or phosphoenolpyruvate carboxykinase-mediated conversion of oxaloacetate to phosphoenolpyruvate and subsequent hydrolysis of the latter to pyruvate, and introduction of the labeled pyruvate into the TCA cycle, i.e., after exit of the carbon skeleton of pyruvate from the TCA cycle followed by re-entry of the same pyruvate molecules via acetyl CoA. In agreement with earlier observations, pyruvate recycling was demonstrated in astrocytes, indic...
GABA, which is present in the brain in large amounts, is distributed among distinctly different c... more GABA, which is present in the brain in large amounts, is distributed among distinctly different cellular pools, possibly reflecting its multiple functions as metabolite, neurotransmitter, and neurotrophin. Its metabolic enzymes also exhibit heterogeneity, because glutamate decarboxylase exists in two isoforms with different subcellular distribution and regulatory properties. Moreover, recent evidence points to a more pronounced regulatory role of the tricarboxylic acid cycle than hitherto anticipated in the biosynthetic machinery responsible for formation of GABA from glutamine. Additionally, GABAergic neurons may contain distinct populations of mitochondria having different turnover rates of the tricarboxylic acid cycle with different levels of association with GABA synthesis from 2-oxoglutarate via glutamate. These aspects are discussed in relation to the different functional roles of GABA and its prominent involvement in epileptogenic activity.
In patients with impaired liver function and hepatic encephalopathy (HE), consistent elevations o... more In patients with impaired liver function and hepatic encephalopathy (HE), consistent elevations of blood ammonia concentration suggest a crucial role in the pathogenesis of HE. Ammonia and acetate are metabolized in brain both primarily in astrocytes. Here, we used dynamic [(11)C]acetate PET of the brain to measure the contribution of astrocytes to the previously observed reduction of brain oxidative metabolism in patients with liver cirrhosis and HE, compared to patients with cirrhosis without HE, and to healthy subjects. We used a new kinetic model to estimate uptake from blood to astrocytes and astrocyte metabolism of [(11)C]acetate. No significant differences of the rate constant of oxidation of [(11)C]acetate (k 3) were found among the three groups of subjects. The net metabolic clearance of [(11)C]acetate from blood was lower in the group of patients with cirrhosis and HE than in the group of healthy subjects (P < 0.05), which we interpret to be an effect of reduced cerebra...
The fine tuning of both glutamatergic and GABAergic neurotransmission is to a large extent depend... more The fine tuning of both glutamatergic and GABAergic neurotransmission is to a large extent dependent upon optimal function of astrocytic transport processes. Thus, glutamate transport in astrocytes is mandatory to maintain extrasynaptic glutamate levels sufficiently low to prevent excitotoxic neuronal damage. In GABA synapses hyperactivity of astroglial GABA uptake may lead to diminished GABAergic inhibitory activity resulting in seizures. As a consequence of this the expression and functional activity of astrocytic glutamate and GABA transport is regulated in a number of ways at transcriptional, translational and post-translational levels. This opens for a number of therapeutic strategies by which the efficacy of excitatory and inhibitory neurotransmission may be manipulated.
The classical concept regarding compartmentation of brain metabolism pertinent to the two neurotr... more The classical concept regarding compartmentation of brain metabolism pertinent to the two neurotransmitter amino acids, glutamate and GABA (gamma-aminobutyrate), operates with different pools of glutamate and glutamine in different cell types, that is, pools that have different sizes and turnover rates. As a result of more sophisticated technology (e.g., nuclear magnetic resonance spectroscopy and mass spectrometry used in relation to cultured neurons and astrocytes), a more complex scenario is emerging. Hence, both neurons and astrocytes exhibit a compartmentalized metabolism that very likely relates to individual cells containing mitochondrial populations having different metabolic roles. Models for this in neurons and astrocytes, respectively, are presented. The functional implications of this for the homeostatic mechanisms regulating the levels of neurotransmitter glutamate and GABA are discussed in relation to development of therapeutic strategies for neurological disorders in which these transmitters are believed to play important roles.
Interaction Between Neurons and Glia in Aging and Disease, 2007
The functional activity in the brain is primarily composed of interplay between excitation and in... more The functional activity in the brain is primarily composed of interplay between excitation and inhibition. In any given region the output is based upon a complex processing of incoming signals that require both excitatory and inhibitory units. Moreover, these units must be regulated and balanced such that an integrated and finely tuned response is generated. In each of these units
Diseases of the brain account for much human suffering and place a huge burden on the health care... more Diseases of the brain account for much human suffering and place a huge burden on the health care systems. Thus, research into the pathology of brain diseases and improved pharmacotherapy is of significant value. In this respect, knowledge on malfunctions of metabolic homeostasis related to the neurotransmission process is still limited. As evident from this chapter, failure of the metabolic
Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism, 2014
Despite the well-established use of kainate as a model for seizure activity and temporal lobe epi... more Despite the well-established use of kainate as a model for seizure activity and temporal lobe epilepsy, most studies have been performed at doses giving rise to general limbic seizures and have mainly focused on neuronal function. Little is known about the effect of lower doses of kainate on cerebral metabolism and particularly that associated with astrocytes. We investigated astrocytic and neuronal metabolism in the cerebral cortex of adult mice after treatment with saline (controls), a subconvulsive or a mildly convulsive dose of kainate. A combination of [1,2-(13)C]acetate and [1-(13)C]glucose was injected and subsequent nuclear magnetic resonance spectroscopy of cortical extracts was employed to distinctively map astrocytic and neuronal metabolism. The subconvulsive dose of kainate led to an instantaneous increase in the cortical lactate content, a subsequent reduction in the amount of [4,5-(13)C]glutamine and an increase in the calculated astrocytic TCA cycle activity. In contr...
Diet supplementation with ketone bodies (acetoacetate and β-hydroxybuturate) or medium-length fat... more Diet supplementation with ketone bodies (acetoacetate and β-hydroxybuturate) or medium-length fatty acids generating ketone bodies has consistently been found to cause modest improvement of mental function in Alzheimer's patients. It was suggested that the therapeutic effect might be more pronounced if treatment was begun at a preclinical stage of the disease instead of well after its manifestation. The preclinical stage is characterized by decade-long glucose hypometabolism in brain, but ketone body metabolism is intact even initially after disease manifestation. One reason for the impaired glucose metabolism may be early destruction of the noradrenergic brain stem nucleus, locus coeruleus, which stimulates glucose metabolism, at least in astrocytes. These glial cells are essential in Alzheimer pathogenesis. The β-amyloid peptide Aβ interferes with their cholinergic innervation, which impairs synaptic function due to diminished astrocytic glutamate release. Aβ also reduces gluc...
Cultured astrocytes treated with siRNA to knock down glutamate dehydrogenase (GDH) were used to i... more Cultured astrocytes treated with siRNA to knock down glutamate dehydrogenase (GDH) were used to investigate whether this enzyme is important for the utilization of glutamate as an energy substrate. By incubation of these cells in media containing different concentrations of glutamate (range 100-500 µM) in the presence or in the absence of glucose, the metabolism of these substrates was studied by using tritiated glutamate or 2-deoxyglucose as tracers. In addition, the cellular contents of glutamate and ATP were determined. The astrocytes were able to maintain physiological levels of ATP regardless of the expression level of GDH and the incubation condition, indicating a high degree of flexibility with regard to regulatory mechanisms involved in maintaining an adequate energy level in the cells. Glutamate uptake was found to be increased in these cells when exposed to increasing levels of extracellular glutamate independently of the GDH expression level. Moreover, increased intracell...
The central nervous system (CNS) operates by a fine-tuned balance between excitatory and inhibito... more The central nervous system (CNS) operates by a fine-tuned balance between excitatory and inhibitory signalling. In this context, the inhibitory neurotransmission may be of particular interest as it has been suggested that such neuronal pathways may constitute 'command pathways' and the principle of 'dis-inhibition' leading ultimately to excitation may play a fundamental role (Roberts, E. (1974). Adv. Neurol., 5: 127-143). The neurotransmitter responsible for this signalling is gamma-aminobutyrate (GABA) which was first discovered in the CNS as a curious amino acid (Roberts, E., Frankel, S. (1950). J. Biol. Chem., 187: 55-63) and later proposed as an inhibitory neurotransmitter (Curtis, D.R., Watkins, J.C. (1960). J. Neurochem., 6: 117-141; Krnjevic, K., Schwartz, S. (1967). Exp. Brain Res., 3: 320-336). The present review will describe aspects of GABAergic neurotransmission related to homeostatic mechanisms such as biosynthesis, metabolism, release and inactivation. ...
Pyruvate recycling was studied in primary cultures of mouse cerebrocortical astrocytes, GABAergic... more Pyruvate recycling was studied in primary cultures of mouse cerebrocortical astrocytes, GABAergic cerebrocortical interneurons, and co-cultures consisting of both cell types by measuring production of [4-(13)C]glutamate from [3-(13)C]glutamate by aid of nuclear magnetic resonance spectroscopy. This change in the position of the label can only occur by entry of [3-(13)C]glutamate into the tricarboxylic acid (TCA) cycle, conversion of labeled alpha-ketoglutarate to malate or oxaloacetate, malic enzyme-mediated decarboxylation of malate to pyruvate or phosphoenolpyruvate carboxykinase-mediated conversion of oxaloacetate to phosphoenolpyruvate and subsequent hydrolysis of the latter to pyruvate, and introduction of the labeled pyruvate into the TCA cycle, i.e., after exit of the carbon skeleton of pyruvate from the TCA cycle followed by re-entry of the same pyruvate molecules via acetyl CoA. In agreement with earlier observations, pyruvate recycling was demonstrated in astrocytes, indic...
GABA, which is present in the brain in large amounts, is distributed among distinctly different c... more GABA, which is present in the brain in large amounts, is distributed among distinctly different cellular pools, possibly reflecting its multiple functions as metabolite, neurotransmitter, and neurotrophin. Its metabolic enzymes also exhibit heterogeneity, because glutamate decarboxylase exists in two isoforms with different subcellular distribution and regulatory properties. Moreover, recent evidence points to a more pronounced regulatory role of the tricarboxylic acid cycle than hitherto anticipated in the biosynthetic machinery responsible for formation of GABA from glutamine. Additionally, GABAergic neurons may contain distinct populations of mitochondria having different turnover rates of the tricarboxylic acid cycle with different levels of association with GABA synthesis from 2-oxoglutarate via glutamate. These aspects are discussed in relation to the different functional roles of GABA and its prominent involvement in epileptogenic activity.
In patients with impaired liver function and hepatic encephalopathy (HE), consistent elevations o... more In patients with impaired liver function and hepatic encephalopathy (HE), consistent elevations of blood ammonia concentration suggest a crucial role in the pathogenesis of HE. Ammonia and acetate are metabolized in brain both primarily in astrocytes. Here, we used dynamic [(11)C]acetate PET of the brain to measure the contribution of astrocytes to the previously observed reduction of brain oxidative metabolism in patients with liver cirrhosis and HE, compared to patients with cirrhosis without HE, and to healthy subjects. We used a new kinetic model to estimate uptake from blood to astrocytes and astrocyte metabolism of [(11)C]acetate. No significant differences of the rate constant of oxidation of [(11)C]acetate (k 3) were found among the three groups of subjects. The net metabolic clearance of [(11)C]acetate from blood was lower in the group of patients with cirrhosis and HE than in the group of healthy subjects (P < 0.05), which we interpret to be an effect of reduced cerebra...
The fine tuning of both glutamatergic and GABAergic neurotransmission is to a large extent depend... more The fine tuning of both glutamatergic and GABAergic neurotransmission is to a large extent dependent upon optimal function of astrocytic transport processes. Thus, glutamate transport in astrocytes is mandatory to maintain extrasynaptic glutamate levels sufficiently low to prevent excitotoxic neuronal damage. In GABA synapses hyperactivity of astroglial GABA uptake may lead to diminished GABAergic inhibitory activity resulting in seizures. As a consequence of this the expression and functional activity of astrocytic glutamate and GABA transport is regulated in a number of ways at transcriptional, translational and post-translational levels. This opens for a number of therapeutic strategies by which the efficacy of excitatory and inhibitory neurotransmission may be manipulated.
The classical concept regarding compartmentation of brain metabolism pertinent to the two neurotr... more The classical concept regarding compartmentation of brain metabolism pertinent to the two neurotransmitter amino acids, glutamate and GABA (gamma-aminobutyrate), operates with different pools of glutamate and glutamine in different cell types, that is, pools that have different sizes and turnover rates. As a result of more sophisticated technology (e.g., nuclear magnetic resonance spectroscopy and mass spectrometry used in relation to cultured neurons and astrocytes), a more complex scenario is emerging. Hence, both neurons and astrocytes exhibit a compartmentalized metabolism that very likely relates to individual cells containing mitochondrial populations having different metabolic roles. Models for this in neurons and astrocytes, respectively, are presented. The functional implications of this for the homeostatic mechanisms regulating the levels of neurotransmitter glutamate and GABA are discussed in relation to development of therapeutic strategies for neurological disorders in which these transmitters are believed to play important roles.
Uploads
Papers by Helle Waagepetersen