Astrocyte influences on ischemic neuronal death

Glutamate excitotoxicity, oxidative stress, and acidosis are primary mediators of neuronal death during ischemia and reperfusion. Astrocytes influence these processes in several ways. Glutamate uptake by astrocytes normally prevents excitotoxic glutamate elevations in brain extracellular space, and this process appears to be a critical determinant of neuronal survival in the ischemic penumbra. Conversely, glutamate efflux from astrocytes by reversal of glutamate uptake, volume sensitive organic ion channels, and other routes may contribute to extracellular glutamate elevations. Glutamate activation of neuronal N-methyl-D-aspartate (NMDA) receptors is modulated by glycine and D-serine: both of these neuromodulators are transported by astrocytes, and D-serine production is localized exclusively to astrocytes. Astrocytes influence neuronal antioxidant status through release of ascorbate and uptake of its oxidized form, dehydroascorbate, and by indirectly supporting neuronal glutathione metabolism. In addition, glutathione in astrocytes can serve as a sink for nitric oxide and thereby reduce neuronal oxidant stress during ischemia. Astrocytes probably also influence neuronal survival in the post-ischemic period. Reactive astrocytes secrete nitric oxide, TNFalpha, matrix metalloproteinases, and other factors that can contribute to delayed neuronal death, and facilitate brain edema via aquaporin-4 channels localized to the astrocyte endfoot-endothelial interface. On the other hand erythropoietin, a paracrine messenger in brain, is produced by astrocytes and upregulated after ischemia. Erythropoietin stimulates the Janus kinase-2 (JAK-2) and nuclear factor-kappaB (NF-kB) signaling pathways in neurons to prevent programmed cell death after ischemic or excitotoxic stress. Astrocytes also secrete several angiogenic and neurotrophic factors that are important for vascular and neuronal regeneration after stroke.     

Astrocyte–neuron interactions in neurological disorders

Astrocytes have long been considered as just providing trophic support for neurons in the central nervous system, but recently several studies have highlighted their importance in many functions such as neurotransmission, metabolite and electrolyte homeostasis, cell signaling, inflammation, and synapse modulation. Astrocytes are, in fact, part of a bidirectional crosstalk with neurons. Moreover, increasing evidence is stressing the emerging role of astrocyte dysfunction in the pathophysiology of neurological disorders, including neurodegenerative disease, stroke, epilepsy, migraine, and neuroinflammatory diseases.     

A Commentary on the Need for 3D-Biologically Relevant In Vitro Environments to Investigate Astrocytes and Their Role in Central Nervous System Inflammation

Astrocytes execute essential functions in the healthy CNS, whilst also being implicated as a limitation to functional regeneration and repair after injury. They respond to injury to minimize damage to healthy tissue whilst also attempting to seal the broken blood-brain-barrier, however, they impede recovery if they are persistent and form a permanent scar in the injured brain. As such, it is of great importance to understand the mechanism underlying the astrocytic response to injury, and this understanding is currently limited by the in vitro environments available to scientists. Biomaterials such as nanofibres and hydrogels offer great potential for the development of superior, 3D cell culture environments in which to study astrocyte behavior and phenotype. The implementation of such in vitro environments with a particularly interdisciplinary approach can improve the field’s understanding of astrocytes, their role in central nervous system inflammation, and elucidate potential strategies to achieve functional regeneration.     

Role of PACAP and VIP in astroglial functions

Astrocytes represent at least 50% of the volume of the human brain. Besides their roles in various supportive functions, astrocytes are involved in the regulation of stem cell proliferation, synaptic plasticity and neuroprotection. Astrocytes also influence neuronal physiology by responding to neurotransmitters and neuropeptides and by releasing regulatory factors termed gliotransmitters. In particular, astrocytes express the PACAP-specific receptor PAC1-R and the PACAP/VIP mutual receptors VPAC1-R and VPAC2-R during development and/or in the adult. There is now clear evidence that PACAP and VIP modulate a number of astrocyte activities such as proliferation, plasticity, glycogen production, and biosynthesis of neurotrophic factors and gliotransmitters.     

Astrocyte Mitochondria in In Vitro Models of Ischemia

There is growing evidence that preservation of mitochondrial respiratory function during cerebral ischemia-reperfusion predicts the ultimate extent of tissue injury. Because neurons are selectively vulnerable to ischemic injury, many studies have focused on neuronal mitochondrial dysfunction in ischemia. However, positron emission tomography (PET) studies in animals and humans suggest that non-neuronal cells such as astrocytes may also experience mitochondrial metabolic compromise that contributes to ischemic necrosis. Astrocytes carry out a number of functions that are critical to normal nervous system function, including uptake of neurotransmitters, regulation of pH and ion concentrations, and metabolic support of neurons. Mitochondria are important for many of these actions. We have used a cell culture model of stroke, oxygen-glucose deprivation (OGD), to study the response of astrocyte mitochondria to ischemia, and to evaluate how changes in astrocyte mitochondrial function might affect neuronal survival and recovery after ischemia.     

Optimized and efficient preparation of astrocyte cultures from rat spinal cord

Astrocytes constitute a major class of glial cells in the CNS, and play crucial roles in physiological functioning, performance and maintenance of the CNS, as well as promotion of neuronal migration and maturation. Astrocytes have also been directly and indirectly implicated in the pathophysiology of various trauma occurrences, development of neurodegenerative diseases and nerve regeneration. To further understand mechanisms by which astrocytes elicit these effects, the first critical step in the study of astrocytes is the preparation of purified astrocytes cultures. Here we describe a simple and convenient procedure for producing rat primary astrocyte cultures of high purity, viability and proliferation. For astrocyte culture, we have optimized the isolation procedures and cultivation conditions including coating substrates, enzyme digestion, seeding density and composition of the culture medium. Using immunofluorescent antibodies against GFAP and OX-42 in combination of Hoechst 33342 fluorescent staining, we found that the purity of the astrocyte cultures was >99%. Astrocytes had high viability as measured by 3-(4, 5-dimethyl-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. In addition, flow cytometric analysis was used to measure and observe variations in the cell cycle after 1–2 passages and proliferation of astrocytes was detected with a high percentage of cells stand in S+G₂/M phase. Therefore, the method described here is ideal for experiments, which require highly pure astrocyte cultures.     

Experiments and Observations on Regeneration in Myriothela cocksi

Myriothela cocksi. a solitary athecate hydropolyp showing many peculiarities and being well investigated concerning gonophore development and embryology, has up to now hardly been examined in respect to regeneration. The simple experiments performed here demonstrate the capacity of all three body regions to regenerate, in due time, previously amputated parts entirely. Further, they reveal a clearly two-phased regeneration process. The first phase leads to a repaired wound, the second results in a rebuilding of the missing part. Ecto- and endoderm are differently involved in regeneration. A greater importance of the endoderm for the first stages of regeneration has been found. Finally, the experiments prove an existing polarity in the polyp. On the basis of these findings it can be concluded that the polyp meets the major criteria of regeneration. This is an absolutely necessary presupposition for all further experiments.     

Targeted Activation of Astrocytes: A Potential Neuroprotective Strategy

Astrocytes are involved in many key physiological processes in the brain, including glutamatergic transmission, energy metabolism, and blood flow control. They become reactive in response to pathological situations, a response that involves well-described morphological alterations and less characterized functional changes. The functional consequences of astrocyte reactivity seem to depend on the molecular pathway involved and may result in the enhancement of several neuroprotective and neurotrophic functions. We propose that a selective and controlled activation of astrocytes may switch these highly pleiotropic cells into therapeutic agents to promote neuron survival and recovery. This may represent a potent therapeutic strategy for many brain diseases in which neurons would benefit from an increased support from activated astrocytes.     

Effects of carboxylic acids on the uptake of non-transferrin-bound iron by astrocytes

The concentrations of non-transferrin-bound iron are elevated in the brain during pathological conditions such as stroke and Alzheimer's disease. Astrocytes are specialised for sequestering this iron, however little is known about the mechanisms involved. Carboxylates, such as citrate, have been reported to facilitate iron uptake by intestinal cells. Citrate binds iron and limits its redox activity. The presence of high citrate concentrations in the interstitial fluid of the brain suggests that citrate may be an important ligand for iron transport by astrocytes. This study investigates whether iron accumulation by cultured rat astrocytes is facilitated by citrate or other carboxylates. Contrary to expectations, citrate, tartrate and malate were found to block iron accumulation in a concentration-dependent manner; α-ketoglutarate had limited effects, while fumarate, succinate and glutarate had no effect. This blockade was not due to an inhibition of ferric reductase activity. Instead, it appeared to be related to the capacity of these carboxylates to bind iron, since phosphate, which also binds iron, diminished the capacity of citrate, tartrate and malate to block the cellular accumulation of iron. These findings raise the possibility that citrate may have therapeutic potential in the management of neurodegenerative conditions that involve cellular iron overload.     

Unusual testicular accessory organs, the testicular blind pouches of blennies (Teleostei, Blenniidae). Fine structure, (enzyme-) histochemistry and possible functions

The testicular blind pouches (TBPs) of Lipophrys canevae and Solaria pavo have an annual reproductive cycle with an active secretion phase during spawning, a regression phase correlated with necrotic processes during post-spawning, an inactive regeneration phase during inter-spawning and prespawning. As revealed by fine structure and enzyme-histochemistry the TBPs of L. canevae and S. pavo are involved in the production of steroid glucuronides, which possibly act as pheromones, Further functions are in the secretion of small amounts of sialomuciri and in lytic activity.