Astroglial Control of Respiratory Rhythm Generating Circuits

Astrocytes, the most numerous glial cells of the central nervous system, are well known to provide neuronal circuits with essential structural and metabolic support. There is also evidence that astrocytes may modulate the activities of neuronal circuits controlling motor rhythms including those of the brainstem’s preBötzinger complex (preBötC) that generates the rhythm of breathing in mammals. However, the extent and mechanisms of active astroglial control of the respiratory rhythm-generating circuits remain unknown. The morphological features of astrocytes in this critical brainstem region are also unknown. In this dissertation, viral gene transfer approaches designed to block or activate astroglial signaling pathways were used to determine the role of preBötC astrocytes in the control of breathing using in vitro and in vivo experimental models. Computer-aided morphometric analyses were used to investigate the structural features of brainstem astrocytes potentially contributing to their functional role. The results from these complementary, multi-faceted experiments show that (i) morphologically, preBötC astrocytes are larger, have more branches, and longer processes when compared to astrocytes residing in other regions of the brainstem; (ii) in conscious adult rats, blockade of vesicular release mechanisms or ATP-mediated signaling in preBötC astrocytes by virally-induced bilateral expression of either the light chain of tetanus toxin (TeLC), the dominant-negative SNARE proteins (dnSNARE), or a potent ectonucleotidase – transmembrane prostatic acid phosphatase – results in a significant reduction of resting respiratory frequency and frequency of sighs, augmented breaths that engage preBötC circuits to increase inspiratory effort; (iii) hypoxic- and CO2-induced ventilatory responses are significantly reduced when vesicular release mechanisms in preBötC astrocytes are blocked; (iv) activation of preBötC astrocytes expressing Gq-coupled Designer Receptor Exclusively Activated by Designer Drug is associated with higher frequency of both normal inspirations and sighs; (v) blockade of vesicular release mechanisms (expression of TeLC or dnSNARE) in preBötC astrocytes is associated with a dramatic reduction of exercise capacity. These data suggest that astroglial mechanisms involving exocytotic vesicular release of signaling molecules (gliotransmitters), provides tonic excitatory drive to the inspiratory rhythm-generating circuits of the preBötC and contributes to the generation of sighs. The role of preBötC astrocytes in central nervous mechanisms controlling breathing becomes especially important in conditions of metabolic stress requiring homeostatic adjustments of breathing such as systemic hypoxia, hypercapnia, and exercise, when enhanced respiratory efforts are critical to support physiological and behavioral demands of the body

The power of the business media: Evidence from firm-level productivity

We examine the impact of media coverage on firm-level productivity and find that firms with higher media coverage are associated with higher productivity. Using the launch of Barron\u27s Online as a quasi-shock to media coverage, we document that this relationship is causal. Further exploration shows that the positive media–productivity relationship is stronger for firms with weaker governance mechanisms and for those with higher levels of information asymmetry. We also identify an increase in reputational and career concerns and a reduction in managerial shirking as channels through which media coverage affects firm-level productivity. The results are robust to alternative explanations and endogeneity concerns. Overall, our findings suggest that media coverage reduces managerial opportunism, and thus enhances resource deployment decisions

Analysis of Soil Nailed Walls Under Harmonic Dynamic Excitations Using Finite Difference Method

Soil nailing is an efficient method to stabilize different soil structures. The method has been extensively used for improving stability of slopes. The construction process of Soil nailed walls commonly involve three basic sections: excavation, nail installation and face stabilization. The nail bars are inserted into ground by either drilling or grouting and are usually arranged in both horizontal and vertical directions. Present research intends to understand Soil-nailed wall behavior under dynamic excitations. Employing finite difference method a three dimensional model has been developed in the proper finite difference code. Soil constitutive behavior for dynamic analyses is predicted taking into account soil hysteresis behavior. To simulate nail bars cable structural elements are employed and also liner structural elements will be utilized for shotcrete facing. Dynamic excitation incorporated as semi-seismic harmonic loading is applied at the bottom of the model where represents soil subgrade. The boundary conditions are considered to be antisymmetric during dynamic analyses. Effects of different crucial factors are monitored during investigations. Some parameters such as, input motion frequency, nail inclination, nail length as well as soil strength properties have been examined

Synuclein Deficiency Results in Age-Related Respiratory and Cardiovascular Dysfunctions in Mice

Synuclein (α, β, and γ) proteins are highly expressed in presynaptic terminals, and significant data exist supporting their role in regulating neurotransmitter release. Targeting the gene encoding α-synuclein is the basis of many animal models of Parkinson’s disease (PD). However, the physiological role of this family of proteins in not well understood and could be especially relevant as interfering with accumulation of α-synuclein level has therapeutic potential in limiting PD progression. The long-term effects of their removal are unknown and given the complex pathophysiology of PD, could exacerbate other clinical features of the disease, for example dysautonomia. In the present study, we sought to characterize the autonomic phenotypes of mice lacking all synucleins (α, β, and γ; αβγ−/−) in order to better understand the role of synuclein-family proteins in autonomic function. We probed respiratory and cardiovascular reflexes in conscious and anesthetized, young (4 months) and aged (18–20 months) αβγ−/− male mice. Aged mice displayed impaired respiratory responses to both hypoxia and hypercapnia when breathing activities were recorded in conscious animals using whole-body plethysmography. These animals were also found to be hypertensive from conscious blood pressure recordings, to have reduced pressor baroreflex gain under anesthesia, and showed reduced termination of both pressor and depressor reflexes. The present data demonstrate the importance of synuclein in the normal function of respiratory and cardiovascular reflexes during aging

The role of parafacial neurons in the control of breathing during exercise

Neuronal cell groups residing within the retrotrapezoid nucleus (RTN) and C1 area of the rostral ventrolateral medulla oblongata contribute to the maintenance of resting respiratory activity and arterial blood pressure, and play an important role in the development of cardiorespiratory responses to metabolic challenges (such as hypercapnia and hypoxia). In rats, acute silencing of neurons within the parafacial region which includes the RTN and the rostral aspect of the C1 circuit (pFRTN/C1), transduced to express HM4D (Gi-coupled) receptors, was found to dramatically reduce exercise capacity (by 60%), determined by an intensity controlled treadmill running test. In a model of simulated exercise (electrical stimulation of the sciatic or femoral nerve in urethane anaesthetised spontaneously breathing rats) silencing of the pFRTN/C1 neurons had no effect on cardiovascular changes, but significantly reduced the respiratory response during steady state exercise. These results identify a neuronal cell group in the lower brainstem which is critically important for the development of the respiratory response to exercise and, determines exercise capacity

Synuclein deficiency results in age-related respiratory and cardiovascular dysfunctions in mice

Synuclein (α, β, and γ) proteins are highly expressed in presynaptic terminals, and significant data exist supporting their role in regulating neurotransmitter release. Targeting the gene encoding α-synuclein is the basis of many animal models of Parkinson’s disease (PD). However, the physiological role of this family of proteins in not well understood and could be especially relevant as interfering with accumulation of α-synuclein level has therapeutic potential in limiting PD progression. The long-term effects of their removal are unknown and given the complex pathophysiology of PD, could exacerbate other clinical features of the disease, for example dysautonomia. In the present study, we sought to characterize the autonomic phenotypes of mice lacking all synucleins (α, β, and γ; αβγ−/−) in order to better understand the role of synuclein-family proteins in autonomic function. We probed respiratory and cardiovascular reflexes in conscious and anesthetized, young (4 months) and aged (18–20 months) αβγ−/− male mice. Aged mice displayed impaired respiratory responses to both hypoxia and hypercapnia when breathing activities were recorded in conscious animals using whole-body plethysmography. These animals were also found to be hypertensive from conscious blood pressure recordings, to have reduced pressor baroreflex gain under anesthesia, and showed reduced termination of both pressor and depressor reflexes. The present data demonstrate the importance of synuclein in the normal function of respiratory and cardiovascular reflexes during aging

Respiratory rhythm irregularity after carotid body denervation in rats

Respiratory activity is controlled by inputs from the peripheral and central chemoreceptors. Since overactivity of the carotid bodies, the main peripheral chemoreceptors, is linked to the pathophysiology of disparate metabolic and cardiovascular diseases, carotid body denervation (CBD) has been proposed as a potential treatment. However, long-term effects of CBD on the respiratory rhythm and regularity of breathing remain unknown. Here, we show that five weeks after bilateral CBD in rats, the respiratory rhythm was slower and less regular. Ten weeks after bilateral CBD, the respiratory frequency was not different from the sham-operated group, but the regularity of the respiratory rhythm was still reduced. Increased frequency of randomly occurring apneas is likely to be responsible for the irregular breathing pattern after CBD. These results should be taken into consideration since any treatment that reduces the stability of the respiratory rhythm might exacerbate the cardio-respiratory instability and worsen the cardiovascular outcomes

Contributions of carotid bodies, retrotrapezoid nucleus neurons and preBötzinger complex astrocytes to the CO2-sensitive drive for breathing

Current models of respiratory CO2 chemosensitivity are centred around the function of a specific population of neurons residing in the medullary retrotrapezoid nucleus (RTN). However, there is significant evidence suggesting that chemosensitive neurons exist in other brainstem areas, including the rhythm-generating region of the medulla oblongata – the preBötzinger complex (preBötC). There is also evidence that astrocytes, non-neuronal brain cells, contribute to central CO2 chemosensitivity. In this study, we reevaluated the relative contributions of the RTN neurons, the preBötC astrocytes, and the carotid body chemoreceptors in mediating the respiratory responses to CO2 in experimental animals (adult laboratory rats). To block astroglial signalling via exocytotic release of transmitters, preBötC astrocytes were targeted to express the tetanus toxin light chain (TeLC). Bilateral expression of TeLC in preBötC astrocytes was associated with ∼20% and ∼30% reduction of the respiratory response to CO2 in conscious and anaesthetized animals, respectively. Carotid body denervation reduced the CO2 respiratory response by ∼25%. Bilateral inhibition of RTN neurons transduced to express Gi-coupled designer receptors exclusively activated by designer drug (DREADDGi) by application of clozapine-N-oxide reduced the CO2 response by ∼20% and ∼40% in conscious and anaesthetized rats, respectively. Combined blockade of astroglial signalling in the preBötC, inhibition of RTN neurons and carotid body denervation reduced the CO2-induced respiratory response by ∼70%. These data further support the hypothesis that the CO2-sensitive drive to breathe requires inputs from the peripheral chemoreceptors and several central chemoreceptor sites. At the preBötC level, astrocytes modulate the activity of the respiratory network in response to CO2, either by relaying chemosensory information (i.e. they act as CO2 sensors) or by enhancing the preBötC network excitability to chemosensory inputs

Brain metabolic sensing and metabolic signaling at the level of an astrocyte

Astrocytes support neuronal function by providing essential structural and nutritional support, neurotransmitter trafficking and recycling and may also contribute to brain information processing. In this article we review published results and report new data suggesting that astrocytes function as versatile metabolic sensors of central nervous system (CNS) milieu and play an important role in the maintenance of brain metabolic homeostasis. We discuss anatomical and functional features of astrocytes that allow them to detect and respond to changes in the brain parenchymal levels of metabolic substrates (oxygen and glucose), and metabolic waste products (carbon dioxide). We report data suggesting that astrocytes are also sensitive to circulating endocrine signals-hormones like ghrelin, glucagon-like peptide-1 and leptin, that have a major impact on the CNS mechanisms controlling food intake and energy balance. We discuss signaling mechanisms that mediate communication between astrocytes and neurons and consider how these mechanisms are recruited by astrocytes activated in response to various metabolic challenges. We review experimental data suggesting that astrocytes modulate the activities of the respiratory and autonomic neuronal networks that ensure adaptive changes in breathing and sympathetic drive in order to support the physiological and behavioral demands of the organism in ever-changing environmental conditions. Finally, we discuss evidence suggesting that altered astroglial function may contribute to the pathogenesis of disparate neurological, respiratory and cardiovascular disorders such as Rett syndrome and systemic arterial hypertension

Functional Oxygen Sensitivity of Astrocytes

In terrestrial mammals, the oxygen storage capacity of the CNS is limited, and neuronal function is rapidly impaired if oxygen supply is interrupted even for a short period of time. However, oxygen tension monitored by the peripheral (arterial) chemoreceptors is not sensitive to regional CNS differences in partial pressure of oxygen (PO2 ) that reflect variable levels of neuronal activity or local tissue hypoxia, pointing to the necessity of a functional brain oxygen sensor. This experimental animal (rats and mice) study shows that astrocytes, the most numerous brain glial cells, are sensitive to physiological changes in PO2 . Astrocytes respond to decreases in PO2 a few millimeters of mercury below normal brain oxygenation with elevations in intracellular calcium ([Ca(2+)]i). The hypoxia sensor of astrocytes resides in the mitochondria in which oxygen is consumed. Physiological decrease in PO2 inhibits astroglial mitochondrial respiration, leading to mitochondrial depolarization, production of free radicals, lipid peroxidation, activation of phospholipase C, IP3 receptors, and release of Ca(2+) from the intracellular stores. Hypoxia-induced [Ca(2+)]i increases in astrocytes trigger fusion of vesicular compartments containing ATP. Blockade of astrocytic signaling by overexpression of ATP-degrading enzymes or targeted astrocyte-specific expression of tetanus toxin light chain (to interfere with vesicular release mechanisms) within the brainstem respiratory rhythm-generating circuits reveals the fundamental physiological role of astroglial oxygen sensitivity; in low-oxygen conditions (environmental hypoxia), this mechanism increases breathing activity even in the absence of peripheral chemoreceptor oxygen sensing. These results demonstrate that astrocytes are functionally specialized CNS oxygen sensors tuned for rapid detection of physiological changes in brain oxygenation. Significance statement: Most, if not all, animal cells possess mechanisms that allow them to detect decreases in oxygen availability leading to slow-timescale, adaptive changes in gene expression and cell physiology. To date, only two types of mammalian cells have been demonstrated to be specialized for rapid functional oxygen sensing: glomus cells of the carotid body (peripheral respiratory chemoreceptors) that stimulate breathing when oxygenation of the arterial blood decreases; and pulmonary arterial smooth muscle cells responsible for hypoxic pulmonary vasoconstriction to limit perfusion of poorly ventilated regions of the lungs. Results of the present study suggest that there is another specialized oxygen-sensitive cell type in the body, the astrocyte, that is tuned for rapid detection of physiological changes in brain oxygenation