Characteristics and Regulation of Glycine Transport in Bergmann Glia

In the vertebrate CNS, glycine acts as an inhibitory neurotransmitter and as the obligatory coagonist of glutamate at N-methyl-d-aspartate receptors. These roles depend on extracellular glycine levels, regulated by Na⁺/Cl⁻-dependent transporters GLYT1, present mainly in glial cells, and GLYT2, predominantly neuronal. In Bergmann glia, GLYT1 mediates both, glycine uptake and efflux, which, in turn, influences excitatory neurotransmission at Purkinje cell synapses. The biochemical properties of GLYTs and their regulation by signaling pathways in these cells are largely unknown. We characterized Gly uptake in confluent primary cultures of Bergmann glia from chick cerebellum. Transport was found to be energy- and Na⁺-dependent, and was resolved into a high (Km=25 μM) and a low affinity (Km=1.1 mM) components identified as GLYT1 and transport System A, respectively. Results show that high affinity transport by GLYT1 is regulated by calcium from intracellular stores, calmodulin, and myosin light chain kinase through an actin cytoskeleton-mediated action. Special issue dedicated to Dr. Simo S. Oja     

Volume interpolation of CT images from tree trunks

Computerized tomography as a non-destructive scanning method to analyze wood structures has become an important technique in tree research. The possibility to reconstruct three-dimensional volumes based on a number of slices of two-dimensional data from CT scans is strongly dependent on the number of measured slices. Radial basis function methods can be successfully used to interpolate CT images with the aim of obtaining a satisfactory reconstruction of tree trunks. In contrast to standard interpolation techniques, our method takes into account that wood structures differ more in the radial than in the longitudinal direction. Therefore we obtain better interpolation results for wood structures.     

Biosynthesis of NAAG by an enzyme-mediated process in rat central nervous system neurons and glia

The peptide transmitter N-acetylaspartylglutamate (NAAG) is present in millimolar concentrations in mammalian spinal cord. Data from the rat peripheral nervous system suggest that this peptide is synthesized enzymatically, a process that would be unique for mammalian neuropeptides. To test this hypothesis in the mammalian CNS, rat spinal cords were acutely isolated and used to study the incorporation of radiolabeled amino acids into NAAG. Consistent with the action of a NAAG synthetase, inhibition of protein synthesis did not affect radiolabel incorporation into NAAG. Depolarization of spinal cords stimulated incorporation of radiolabel. Biosynthesis of NAAG by cortical astrocytes in cell culture was demonstrated by tracing incorporation of [3H]-glutamate by astrocytes. In the first test of the hypothesis that NAA is an immediate precursor in NAAG biosynthesis, [3H]-NAA was incorporated into NAAG by isolated spinal cords and by cell cultures of cortical astrocytes. Data from cerebellar neurons and glia in primary culture confirmed the predominance of neuronal synthesis and glial uptake of NAA, leading to the hypothesis that while neurons synthesize NAA for NAAG biosynthesis, glia may take it up from the extracellular space. However, cortical astrocytes in serum-free low-density cell culture incorporated [3H]-aspartate into NAAG, a result indicating that under some conditions these cells may also synthesize NAA. Pre-incubation of isolated spinal cords and cultures of rat cortical astrocytes with unlabeled NAA increased [3H]-glutamate incorporation into NAAG. In contrast, [3H]-glutamine incorporation in spinal cord was not stimulated by unlabeled NAA. These results are consistent with the glutamate-glutamine cycle greatly favoring uptake of glutamine into neurons and glutamate by glia and suggest that NAA availability may be rate-limiting in the synthesis of NAAG by glia under some conditions.     

Neuronal A1 receptors mediate increase in extracellular kynurenic acid after local intrastriatal adenosine infusion

The naturally occurring purine nucleoside adenosine has pronounced anticonvulsant and neuroprotective properties and plays a neuromodulatory role in the CNS. Kynurenic acid (KYNA) is an astrocyte-derived, endogenous neuroinhibitory compound, which shares several of adenosine's properties. In a first attempt to examine possible interactions between these two biologically active molecules, adenosine was focally applied into the striatum of freely moving rats by reverse microdialysis, and changes in extracellular KYNA were monitored over time. A 2-h infusion of adenosine increased KYNA levels in a dose-dependent manner, with 10 mm of adenosine causing a twofold elevation within 1 h. This effect was reversible and was effectively blocked by coinfusion of the specific A1 adenosine receptor antagonist 8-cyclopentyltheophylline (100 microm). In contrast, coinfusion of adenosine with MSX-3 (100 microm), an A2A receptor antagonist, did not affect the adenosine-induced increase in KYNA levels. Local striatal perfusion with the A1 receptor agonist N6-cyclopentyladenosine (100 microm) mimicked the effect of adenosine, whereas perfusion with the A2A receptor agonist CGS-21680 (100 microm) was ineffective. Finally, we tested the effect of adenosine (10 mm) on extracellular KYNA in striata that had been injected with quinolinate (60 nmol/1 microL) 7 days earlier. In this neuron-depleted tissue, perfusion with adenosine failed to affect extracellular KYNA levels. These data demonstrate that adenosine is capable of raising extracellular KYNA in the rat striatum by interacting with postsynaptic neuronal A1 receptors. This mechanism may result in a synergism between the neurobiological effects of adenosine and KYNA.     

Sapling growth and survivorship as affected by light and flooding in a river floodplain forest of southeast Texas

We investigated the effects of light and flooding on growth and survivorship of saplings in a river floodplain forest of southeast Texas. Growth responses to light were consistent with the expectation that shade-intolerant species grow faster than shade-tolerant species in high light, and vice versa. Mortality risk was not associated with shade tolerance level unless high mortality risks associated with a period of high flooding were removed. These results support the hypothesis that shade-tolerant species in floodplains may be limited by flooding as previous studies suggested. Also, compared to their performance at a nearby mesic site, common species showed little intraspecific difference in shade tolerance, especially for shade-intolerant species. Finally, the positive correlation between low-light growth and survivorship suggests that carbon allocation to continued growth may be favored as a sapling strategy in floodplains.     

Neuronal-glial networks as substrate for CNS integration

Astrocytes have been considered, for a long time, as the support and house-keeping cells of the nervous system. Indeed, the astrocytes play very important metabolic roles in the brain, but the catalogue of nervous system functions or activities that involve directly glial participation has extended dramatically in the last decade. In addition to the further refining of the signalling capacity of the neuroglial networks and the detailed reassessment of the interactions between glia and vascular bed in the brain, one of the important salient features of the increased glioscience activity in the last few years was the morphological and functional demonstration that protoplasmic astrocytes occupy well defined spatial territories, with only limited areas of morphological overlapping, but still able to communicate with adjacent neighbours through intercellular junctions. All these features form the basis for a possible reassessment of the nature of integration of activity in the central nervous system that could raise glia to a role of central integrator.     

Inwardly rectifying potassium channels (Kir) in central nervous system glia: a special role for Kir4.1 in glial functions

Glia in the central nervous system (CNS) express diverse inward rectifying potassium channels (Kir). The major function of Kir is in establishing the high potassium (K+) selectivity of the glial cell membrane and strongly negative resting membrane potential (RMP), which are characteristic physiological properties of glia. The classical property of Kir is that K+ flows inwards when the RMP is negative to the equilibrium potential for K+ (E(K)), but at more positive potentials outward currents are inhibited. This provides the driving force for glial uptake of K+ released during neuronal activity, by the processes of "K+ spatial buffering" and "K+ siphoning", considered a key function of astrocytes, the main glial cell type in the CNS. Glia express multiple Kir channel subtypes, which are likely to have distinct functional roles related to their differences in conductance, and sensitivity to intracellular and extracellular factors, including pH, ATP, G-proteins, neurotransmitters and hormones. A feature of CNS glia is their specific expression of the Kir4.1 subtype, which is a major K+ conductance in glial cell membranes and has a key role in setting the glial RMP. It is proposed that Kir4.1 have a primary function in K+ regulation, both as homomeric channels and as heteromeric channels by co-assembly with Kir5.1 and probably Kir2.0 subtypes. Significantly, Kir4.1 are also expressed by oligodendrocytes, the myelin-forming cells of the CNS, and the genetic ablation of Kir4.1 results in severe hypomyelination. Hence, Kir, and in particular Kir4.1, are key regulators of glial functions, which in turn determine neuronal excitability and axonal conduction.     

四种挺水植物生理生态特性和污水净化效果研究

采用人工气候室水培系统以人工污水培养慈姑(Sagittaria trifolia)、花皇冠(Echinodorus berteroi)、菖蒲(Acorus calamus)和芦苇(Phragmites australis) 4种挺水植物,比较它们的根和地上部分生物量、根长、根寿命、根孔隙度、根径向泌氧量(ROL)、光合作用等生理生态特性及对总氮(TN)、总磷(TP)、化学需氧量(COD)的去除效果。结果表明,ROL与根孔隙度、光合速率、地上生物量呈显著正相关(P<0.05),与根长极显著正相关(P<0.01);TP的去除与光合速率、COD的去除与ROL显著正相关; TN的去除与生物量极显著正相关(P<0.01),但与根生物量和地上部分生物量的比值(根茎比)显著负相关(P<0.05)。慈姑和花皇冠拥有庞大生物量和发达的根系,根孔隙度、ROL和光合作用等生理指标较高,在水培系统中的污水净化效果接近甚至优于菖蒲和芦苇,是构建人工湿地的优良植物。