Kallikrein-family serine protease in the central nervous system

Serine proteases exert a variety of functions under physiological and pathological conditions. Tissue plasminogen activator (tPA) is expressed widely in the central nervous system (CNS) and play important roles in development, synaptic plasticity and neuronal cell death. In addition to this protease, recent studies have revealed the existence of new serine proteases in the CNS. In particular, two members of the kallikrein gene family, KLK8/neuropsin and KLK6/protease M/neurosin/zyme are expressed abundantly in the CNS. Neuropsin is expressed by the neurons of the hippocampal subfields CA1 and CA3 and shown to cleave extracellular proteins such as fibronectin and L1. This protease plays essential roles in synaptic plasticity such as long-term potentiation (LTP) and kindling. Application of recombinant neuropsin significantly promoted LTP induction and anti-neuropsin antibody reduced potentiation. Intraventricular administration of anti-neuropsin antibody ameliorated kindling epilepsy. Neuropsin-knockout mice (neuropsin-KO) had significantly smaller number of synapses in the CA1 subfield of the hippocampus. These data suggest that neuropsin plays an important role in synapse formation through modifying extracellular environments. After injury to the CNS, neuropsin is expressed in oligodendrocytes around the lesion. Myelins in the severed optic nerve of neuropsin-KO were more preserved than those of wild-type mice, suggesting that neuropsin after injury is involved in myelin degradation. Another kallikrein member, protease M is constitutively expressed in the oligodendrocytes. Insult to the CNS increases protease M expression not only in the oligodendrocytes but also in the inflammatory cells such as macrophages. These proteases in balance with inhibitors are implicated in the modulation of the extracellular environment.     

The role of astrocytes in the physiology and pathology of the central nervous system

Astrocytes are the main class of neuroglia, serving a wide range of adaptive functions in the mammalian nervous system. They interact with neurons, providing structural, metabolic and trophic support for them. In pathological circumstances, astrocytes have the potential to induce neuronal dysfunction, but they can also play a neuroprotective role, releasing neuronal growth factors. Here we review recent findings regarding the role of astrocytes in the biology of the brain in physiological conditions, as well as their reaction following the onset of neurodegenerative disorders.     

Cdk5: mediator of neuronal development, death and the response to DNA damage

In the central nervous system, cyclin-dependent kinase 5 (Cdk5), an unusual member of the Cdk family, is implicated in the regulation of various physiological processes ranging from neuronal survival, migration and differentiation, to synaptogenesis, synaptic plasticity and neurotransmission. Dysregulation of this kinase has been demonstrated to play a critical role in the pathogenic process of neurodegenerative disorders. DNA damage is emerging as an important pathological component in various neurodegenerative conditions. In this review, we discuss the recent progress regarding the regulation and roles of Cdk5 under physiological conditions, and its dysregulation under pathological conditions, especially in neuronal death mediated by DNA damage.     

Specific activities of individual c-Jun N-terminal kinases in the brain

The c-Jun N-terminal kinases (JNKs) are multifunctional molecules which, on the one hand, regulate various processes in brain development, repair and memory formation. On the other hand, JNKs are potent effectors of neuronal death and neuroinflammation. This review summarizes recent findings on individual JNK functions in the nervous system under pathophysiological conditions and on their regulation by upstream kinases, phosphatases and formation of context-dependent signalosomes. By focusing on different aspects of JNK signaling, it becomes increasingly obvious that the JNK cascade is intricately regulated and intensely dependent on the availability and functionality of its single components and their intracellular localization. Our review also emphasizes, that JNKs are indispensable for neuronal cell death as well as many physiological functions in the brain. Finally, we discuss pharmacological strategies which target pathological JNK activities without affecting their physiological functions.     

Effect of cytokines on neuronal excitability

Numerous studies have shown that proinflammatory cytokines induce or facilitate pain and hyperalgesia in the presence of inflammation, injury to the nervous system or cancer. Besides acting as inflammatory mediators, increasing evidence indicates that cytokines may also specifically interact with receptor and ion channels regulating neuronal excitability, synaptic plasticity and injury under both physiological and pathological conditions. Here we summarize findings on two prototypical proinflammatory cytokines, tumor-necrosis factor-α and interleukin-1β, and their effects on neuronal excitability and ion channels with special regards to pain and hyperalgesia.     

Organization of the nervous system in the model planarian Schmidtea mediterranea: an immunocytochemical study

Freshwater planarians are an emerging model in which to study regeneration at the molecular level. These animals can regenerate a complete central nervous system (CNS) in only a few days. In recent years, hundreds of genes expressed in the nervous system have been identified in two popular planarian species used by several laboratories: Dugesia japonica and Schmidtea mediterranea. Functional analyses of some of those neural genes have allowed the process of CNS regeneration to begin to be elucidated in those animals. However, additional work is required to characterize the different neuronal populations. Thus, the identification or generation of antibodies that act as markers for specific neuronal cell types would be extremely useful not only in obtaining a more detailed characterization of the planarian nervous system but also for the analysis of phenotypes obtained by RNA interference. Here, I have used five different antibodies to describe different neuronal populations in the freshwater planarian S. mediterranea. This study represents a first step in characterizing the organization of the nervous system of this species at the cellular level.     

Immunohistochemical localization of the amino acid transporter SNAT2 in the rat brain

SNAT2 is a neutral amino acid carrier that belongs to the system A family. Since its function in the nervous system remains unclear, we have analyzed its distribution in the rat CNS using specific antisera. Although SNAT2 is expressed widely in the CNS, it is enriched in the spinal cord and the brainstem nuclei, especially those of the auditory system. At the cellular level, SNAT2 was preferentially located in neuronal cell bodies and processes, although it was also strongly expressed in the meninges and ependyma. In astrocytes, the localization of SNAT2 was more restricted since it was intensely expressed in the perivascular end-feet, glia limitans, cerebellar astrocytes and Bergmann glia, but it was less intense in astrocytes of the cerebral parenchyma. Among neurons, the primary sensory neurons of the mesencephalic trigeminal nucleus appeared to be those that most strongly express SNAT2, but many other neurons, including cortical pyramidal cells and their dendrites were also intensely stained. In several regions the transporter was detected in axons, especially in the brainstem, and its presence in both dendrites and axons was confirmed by confocal microscopy and ultrastructural studies. However, while SNAT2 was observed in the large principal dendrites and the small distal dendrites, it was only found in axonal shafts and was excluded from terminals. Some glutamatergic neurons were among the more intensely labeled cells whereas SNAT2 was not detected on GABAergic neurons. The expression of SNAT2 partially coincides with that reported for SNAT1, especially in glutamatergic neurons. Hence, both proteins could fulfill complementary roles in replenishing glutamate pools and be differentially regulated under different physiological conditions. They also seem to co-localize in non-neuronal cells probably contributing to amino acid fluxes through the blood-brain barrier.     

Antiapoptotic and pronecrotic actions of neurotrophins

Neurotrophins promote the differentiation, growth, and survival of neurons in the nervous system. Specifically, neurotrophins promote neuronal survival by interfering with programmed cell death or apoptosis. In addition to roles of neurotrophins as survival factors, neurotrophins can act as risk factors of neuronal injury under various pathological conditions. Neurotrophins markedly potentiate neuronal cell necrosis induced by activation of N-methyl-D-aspartate receptors, deprivation of oxygen and glucose, and free radicals. Moreover, prolonged exposure to neurotrophins results in widespread neuronal necrosis through free radical-mediated mechanisms. Whereas cellular and molecular mechanisms underlying antiapoptosis action of neurotrophins have been well documented, extensive study will be needed to delineate mechanisms for the neurotrophin-induced neuronal necrosis through activation of Trk tyrosine kinase receptors.     

Role of semaphorins in the adult nervous system

In the developing nervous system, extending axons are directed towards their appropriate targets by a myriad of attractive and repulsive guidance cues. Work in the past decade has significantly advanced our understanding of these molecules and has made it increasingly clear that their function is not limited to the guidance of growing axons during embryogenesis. Axon guidance cues fulfill additional roles in angiogenesis, cell migration and the immune system, and often display sustained expression in adulthood. Here we focus on the semaphorin (Sema) family and review their proposed functions in the adult nervous system. Several semaphorin family members continue to be expressed in the adult brain and spinal cord, and increasing evidence indicates that their expression is regulated upon nervous system injury in rodents and in neuropathology in humans. The available evidence suggests that semaphorins might significantly contribute to the maintenance and stability of neuronal networks. Furthermore, semaphorins could play important roles in the regeneration, or failure thereof, of neuronal connections. In the future, genetic manipulation of semaphorins and their receptors in the adult intact and injured nervous system should provide a deeper insight into the mechanisms by which semaphorin signaling contributes to structural plasticity and regeneration in the adult brain.     

Roles for the pro-neurotrophin receptor sortilin in neuronal development, aging and brain injury

Neurotrophins are essential for development and maintenance of the vertebrate nervous system. Paradoxically, although mature neurotrophins promote neuronal survival by binding to tropomyosin receptor kinases and p75 neurotrophin receptor (p75(NTR)), pro-neurotrophins induce apoptosis in cultured neurons by engaging sortilin and p75(NTR) in a death-signaling receptor complex. Substantial amounts of neurotrophins are secreted in pro-form in vivo, yet their physiological significance remains unclear. We generated a sortilin-deficient mouse to examine the contribution of the p75(NTR)/sortilin receptor complex to neuronal viability. In the developing retina, Sortilin 1 (Sort1)(-/-) mice showed reduced neuronal apoptosis that was indistinguishable from that observed in p75(NTR)-deficient (Ngfr(-/-)) mice. To our surprise, although sortilin deficiency did not affect developmentally regulated apoptosis of sympathetic neurons, it did prevent their age-dependent degeneration. Furthermore, in an injury protocol, lesioned corticospinal neurons in Sort1(-/-) mice were protected from death. Thus, the sortilin pathway has distinct roles in pro-neurotrophin-induced apoptotic signaling in pathological conditions, but also in specific stages of neuronal development and aging.     

The astrocyte odyssey

Neurons have long held the spotlight as the central players of the nervous system, but we must remember that we have equal numbers of astrocytes and neurons in the brain. Are these cells only filling up the space and passively nurturing the neurons, or do they also contribute to information transfer and processing? After several years of intense research since the pioneer discovery of astrocytic calcium waves and glutamate release onto neurons in vitro, the neuronal-glial studies have answered many questions thanks to technological advances. However, the definitive in vivo role of astrocytes remains to be addressed. In addition, it is becoming clear that diverse populations of astrocytes coexist with different molecular identities and specialized functions adjusted to their microenvironment, but do they all belong to the umbrella family of astrocytes? One population of astrocytes takes on a new function by displaying both support cell and stem cell characteristics in the neurogenic niches. Here, we define characteristics that classify a cell as an astrocyte under physiological conditions. We will also discuss the well-established and emerging functions of astrocytes with an emphasis on their roles on neuronal activity and as neural stem cells in adult neurogenic zones.     

The astrocyte odyssey

Neurons have long held the spotlight as the central players of the nervous system, but we must remember that we have equal numbers of astrocytes and neurons in the brain. Are these cells only filling up the space and passively nurturing the neurons, or do they also contribute to information transfer and processing? After several years of intense research since the pioneer discovery of astrocytic calcium waves and glutamate release onto neurons in vitro, the neuronal-glial studies have answered many questions thanks to technological advances. However, the definitive in vivo role of astrocytes remains to be addressed. In addition, it is becoming clear that diverse populations of astrocytes coexist with different molecular identities and specialized functions adjusted to their microenvironment, but do they all belong to the umbrella family of astrocytes? One population of astrocytes takes on a new function by displaying both support cell and stem cell characteristics in the neurogenic niches. Here, we define characteristics that classify a cell as an astrocyte under physiological conditions. We will also discuss the well-established and emerging functions of astrocytes with an emphasis on their roles on neuronal activity and as neural stem cells in adult neurogenic zones.     

Expression profile of microRNAs in rat hippocampus following lithium-pilocarpine-induced status epilepticus

Although microRNAs are expressed extensively in the central nervous system in physiological and pathological conditions, their expression in neurological disorder of epilepsy has not been well characterized. Here we investigated microRNA expression pattern in post status epilepticus rats (24h after status). Rat MicroRNA array and differential analysis had detected 19 up-regulated microRNAs and 7 down-regulated microRNAs in rat hippocampus, and four randomly selected deregulated microRNAs (microRNA-34a, microRNA-22, microRNA-125a, microRNA-21) were confirmed by qRT-PCR, then their expression alterations in rat peripheral blood were analyzed. We found that these four deregulated microRNAs were also differentially expressed in rat peripheral blood, and trends for their blood expression alterations were just the same as their counterparts in rat hippocampus. Thus, our results have not only characterized the microRNA expression profile in post status epilepticus rat hippocampus but also demonstrated that some rat hippocampal microRNAs were probably associated with rat peripheral blood microRNAs. Moreover, targets of these deregulated microRNAs were analyzed using bioinformatics and the identified enriched MAPK pathway and long-term potentiation pathway might have been involved in molecular mechanisms concerning neuronal death, inflammation and epileptogenesis.     

Biology of oligodendrocyte and myelin in the mammalian central nervous system

Oligodendrocytes, the myelin-forming cells of the central nervous system (CNS), and astrocytes constitute macroglia. This review deals with the recent progress related to the origin and differentiation of the oligodendrocytes, their relationships to other neural cells, and functional neuroglial interactions under physiological conditions and in demyelinating diseases. One of the problems in studies of the CNS is to find components, i.e., markers, for the identification of the different cells, in intact tissues or cultures. In recent years, specific biochemical, immunological, and molecular markers have been identified. Many components specific to differentiating oligodendrocytes and to myelin are now available to aid their study. Transgenic mice and spontaneous mutants have led to a better understanding of the targets of specific dys- or demyelinating diseases. The best examples are the studies concerning the effects of the mutations affecting the most abundant protein in the central nervous myelin, the proteolipid protein, which lead to dysmyelinating diseases in animals and human (jimpy mutation and Pelizaeus-Merzbacher disease or spastic paraplegia, respectively). Oligodendrocytes, as astrocytes, are able to respond to changes in the cellular and extracellular environment, possibly in relation to a glial network. There is also a remarkable plasticity of the oligodendrocyte lineage, even in the adult with a certain potentiality for myelin repair after experimental demyelination or human diseases.     

Neuronal p38 MAPK signalling: an emerging regulator of cell fate and function in the nervous system

p38 mitogen-activated protein kinases (MAPKs), together with extracellular signal-regulated kinases (ERKs) and c-Jun N-terminal kinases (JNKs), constitute the MAPK family. Multiple intracellular signalling pathways that converge on MAPKs exist in all eukaryotic cells and play pivotal roles in a wide variety of cellular functions. p38 MAPKs and JNKs, also termed stress-activated protein kinases (SAPKs), are preferentially activated by various cytotoxic stresses and cytokines and appear to be potent regulators of stress-induced apoptosis. Whereas JNKs have been shown to play pivotal roles in the regulation of neuronal apoptosis, the role of p38 MAPKs in the nervous system is poorly understood. However, accumulating evidence from mammalian cell culture systems and the strong genetic tool C. elegans suggests that neuronal p38 signalling has diverse functions beyond the control of cell death and survival. This review focuses on possible roles for the p38 pathway in the nervous system, with principal emphasis placed on the roles in neuronal cell fate decision and function.     

Retinoic acids acting through retinoid receptors protect hippocampal neurons from oxygen-glucose deprivation-mediated cell death by inhibition of c-jun-N-terminal kinase and p38 mitogen-activated protein kinase

Retinoic acids (RAs), including all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9-cis RA), play fundamental roles in a variety of physiological events in vertebrates, through their specific nuclear receptors: retinoic acid receptor (RAR) and retinoid X receptor (RXR). Despite the physiological importance of RA, their functional significance under pathological conditions is not well understood. We examined the effect of ATRA on oxygen/glucose-deprivation/reperfusion (OGD/Rep)-induced neuronal damage in cultured rat hippocampal slices, and found that ATRA significantly reduced neuronal death. The cytoprotective effect of ATRA was observed not only in cornu ammonis (CA) 1 but also in CA2 and dentate gyrus (DG), and was attenuated by selective antagonists for RAR or RXR. By contrast, in the CA3 region, no protective effects of ATRA were observed. The OGD/Rep also increased phosphorylated forms of c-jun-N-terminal kinase (P-JNK) and p38 (P-p38) in hippocampus, and specific inhibitors for these kinases protected neurons. ATRA prevented the increases in P-JNK and P-p38 after OGD/Rep, as well as the decrease in NeuN and its shrinkage, all of which were inhibited by antagonists for RAR or RXR. These findings suggest that the ATRA signaling via retinoid receptors results in the inhibition of JNK and p38 activation, leading to the protection of neurons against OGD/Rep-induced damage in the rat hippocampus.     

The neurofilament triplet is present in distinct subpopulations of neurons in the central nervous system of the guinea-pig.

It is commonly assumed that most, if not all, neurons contain the intermediate filament protein class known as the neurofilament protein-triplet. The following study investigated the distribution of neurofilament protein-triplet immunoreactivity in selected regions of the guinea-pig central nervous system using monoclonal antibodies directed against phosphorylation-independent epitopes on the three subunits under optimal tissue processing conditions. Neurofilament protein-triplet immunoreactivity was present in distinct subpopulations of neurons in the cerebellar cortex, neocortex, hippocampal formation, retina, striatum and medulla oblongata. In many of these regions, labelled neurons represented only a small proportion of the total. The selective distribution of this intermediate filament protein class was confirmed in double-labelling experiments using antibodies to the neurofilament protein-triplet in combination with antibodies to other neuronal markers. The distribution of neurofilament protein-triplet immunoreactivity also correlated with the distribution of staining observed with a silver impregnation method based on Bielschowsky. The present results in combination with previous observations have demonstrated that the neurofilament protein-triplet is found in specific subclasses of neurons in different regions of the nervous system. Content of this intermediate filament protein class does not appear to be correlated with neuronal size or length of projection. These results also suggest that the selectivity of staining between neuronal classes observed with classical silver impregnation methods may be due to the presence or absence of the neurofilament protein-triplet. The present results may also provide a new perspective on the basis of the selective vulnerability of neurons in degenerative diseases.     

Haemostasis in oral surgery--the possible pathogenetic implications of oral fibrinolysis on bleeding. Experimental and clinical studies of the haemostatic balance in the oral cavity, with particular reference to patients with acquired and congenital defects of the coagulation system.

Activation and inhibition of the haemostatic system was reviewed including the interaction between the four biological systems involved in haemostasis: the vessel wall, the platelets, the coagulation system and the fibrinolytic system. The haemostatic mechanism is initiated at the site of injury through local activation of surfaces and release of tissue thromboplastin, resulting in formation and deposition of fibrin. The coagulation process is regulated by physiological anticoagulants. Activation of fibrinolysis is triggered by the presence of fibrin, and the role of tissue-type plasminogen activators (t-PA) at the site of fibrin formation in particular is emphasized. The process is regulated by physiological inhibitors, of which alpha 2-antiplasmin, histidine-rich glycoprotein and plasminogen activator inhibitor are reported to be of major physiological significance. The role of fibrinolysis in the regulation of the dynamic haemostatic balance is discussed, elucidated through examples of congenital deficiencies of the coagulation and the fibrinoytic system. Pharmacological inhibitors of fibrinolysis (i.e. epsilon-aminocaproic acid and tranexamic acid) and their possible effect on the haemostatic system are described. The systemic effects on the fibrinolytic system of surgery and oral surgery is reviewed, and it is concluded, that oral surgery has insignificant effects on blood fibrinolysis. In contrast, oral surgery induces changes of fibrinolysis in the oral environment; initially the fibrinolytic activity of saliva is reduced, due to the presence of inhibitors of fibrinolysis originating from the blood and the wound exudate. When bleeding and exudation cease, the fibrinolytic activity of the saliva will increase. Plasminogen and plasminogen activator, identified as t-PA are present in the oral environment under physiological conditions. Plasminogen is secreted in the saliva and the sources of t-PA include oral epithelial cells and gingival crevicular fluid. The presence of plasminogen and t-PA in the oral environment implies that when fibrin is present (i.e. after surgery), fibrinolysis is triggered. Haemorrhagic complications to oral surgery in patients without known defects of the coagulation system is reviewed. It is concluded that the investigations conducted to the present day do not permit final conclusions with respect to the pathophysiological role of defects in the coagulation and the fibrinolytic systems for the development of bleeding after oral surgery. Further investigations are necessary in order to clarify these aspects, and should include extensive laboratory analyses to reveal rare congenital defects such as factor XIII- and alpha 2-antiplasmin deficiencies.(ABSTRACT TRUNCATED AT 400 WORDS)     

Astroglial modulation of neurotransmitter/peptide release from the neurohypophysis: present status

Reviewed in this article are those studies that have contributed heavily to our current conceptualizations of glial participation in the functioning of the magnocellular hypothalamo-neurohypophysial system. This system undergoes remarkable morphological and functional reorganization induced by increased demand for peptide synthesis and release, and this reorganization involves the astrocytic elements in primary roles. Under basal conditions, these glia appear to be vested with the responsibility of controlling the neuronal microenvironment in ways that reduce neuronal excitability, restrict access to neuronal membranes by neuroactive substances and deter neuron neuron interactions within the system. With physiological activation, the glial elements, via receptor-mediated mechanisms, take up new positions. This permissively facilitates neuron neuron interactions such as the exposure of neuronal membranes to released peptides and the formation of gap junctions and new synapses, enhances and prolongs the actions of those excitatory neurotransmitters for which there are glial uptake mechanisms, and facilitates the entry of peptides into the blood. In addition, subpopulations of these glia either newly synthesize or increase synthesis of neuroactive peptides for which their neuronal neighbors have receptors. Release of these peptides by the glia or their functional roles in the system have not yet been demonstrated.