The ventral motoneurone axon bundle in the CNS — a cordone system?

In the developing CNS neighbouring structures are commonly separated by transient barriers termed cordones, some of which coincide with glial elements. Where ventral motoneuron axons cross the spinal white matter as intramedullary bundles to reach the CNS-PNS transitional zone they are surrounded from early development by a glial sleeve resembling a cordone. This becomes better developed with age and, like some cordones, persists into adult life. This could provide a radial conduit which might underlie the capacity of central segments of mature ventral motoneurone axons to regenerate. It may also provide a pathway for glial migration from the central cord to more superficial levels, including the transitional zone, where they help form the CNS-PNS barrier. Axons in the intramedullary bundle and in the surrounding ventral white column mature at different rates. Glial sleeve cells of the intramedullary bundles are apposed to both. Morphometric analysis of the axon-glial relationships of the two populations indicates that glial development proceeds at a different rate in relation to each axon class and that this is influenced by the degree of axonal maturation, which may in turn be related to target contact. Furthermore, early axon glial relationships differ between the two populations. For ventral motoneurone axons these take place in two stages: firstly, glial segregation of axons (resembling that in the PNS) and secondly, oligodendrocytic contact and ensheathment, which leads on to myelination. Axon-glial relationships in the ventral white column begin with the second of these events, as is more typical of early CNS myelination in general.     

Does a structural bridge exist between the DNA and the specialized cytoplasmic organelles during the early part of their development? A mechanism for the positioning of flagella and possibly other cytoplasmic organelles

Cell differentiation involves the development of a new cytoplasm containing a set of specialized organelles such as cilia and flagella which are placed in the cell with a predetermined orientation. Arguments are put forward to show that the orientation of the flagellar apparatus could be brought about by a macromolecular structural bridge between the nucleoid and the assembling flagellar apparatus, the orientation being determined by the spatial geometry inherent in the folding of the DNA. An analysis of differentiation in unicelled eukaryotes suggests that the same basic mechanism of a structural bridge could also apply to the orientation of their cilia and flagella and perhaps may have a more general application in the positioning of cytoplasmic organelles.     

Is astrocyte laminin involved in axon guidance in the mammalian CNS?

This paper provides evidence for the expression of laminin on glia in correlation with axon elongation and nerve pathway formation during embryonic development of the mouse optic nerve and other parts of the central nervous system (CNS). We show that punctate deposits of laminin on immature glial cells precede the entrance of the first optic axons into the nerve, and remain in close association with growing axons. Furthermore, we show that in one particular region of the optic pathway that the retinal ganglion cell axons avoid in normal animals (i.e., the pigmented area of the distal nerve) the punctate laminin matrix is missing. As the optic nerve matures punctate laminin deposits disappear, and laminin is reduced in the astroglial cytoplasm. The close correlation of the punctate form of laminin with early axonal growth is true not only in the optic nerve but also in some other parts of the CNS. We demonstrate such punctate laminin deposits in a model of astrocyte-induced regeneration of the corpus callosum in acallosal mice (G. Smith, R. Miller, and J. Silver, 1986, J. Comp. Neurol. 251, 23-43), and in glia associated with several normal developing axon trajectories, such as the corpus callosum, fornix, and pathways in the embryonic hindbrain. In all of these regions punctate laminin deposits are found on astroglia that are associated with early growing axons. Our results indicate that the punctate form of laminin, produced by astrocytes, may be an important factor involved in axon elongation and nerve pathway formation in the mammalian CNS.     

Ubiquitination in the first cytoplasmic loop of μ-opioid receptors reveals a hierarchical mechanism of lysosomal down-regulation

μ-Type opioid receptors (MORs) are members of the large seven-transmembrane receptor family which transduce the effects of both endogenous neuropeptides and clinically important opioid drugs. Prolonged activation of MORs promotes their proteolytic degradation by endocytic trafficking to lysosomes. This down-regulation process is known to contribute to homeostatic regulation of cellular opioid responsiveness, but mechanisms that mediate and control MOR down-regulation have not been defined. We show here that lysosomal down-regulation of MORs is ESCRT (endosomal sorting complex required for transport)-dependent and involves ubiquitin-promoted transfer of internalized MORs from the limiting endosome membrane to lumen. We also show that MOR down-regulation measured by conventional radioligand binding assay is determined specifically by ubiquitination in the first cytoplasmic loop. Surprisingly, we were unable to find any role of ubiquitination in determining whether internalized receptors recycle or are delivered to lysosomes. Instead, this decision is dictated specifically by the MOR C-tail and occurs irrespectively of the presence or absence of receptor ubiquitination. Our results support a hierarchical organization of discrete ubiquitin-independent and -dependent sorting operations, which function non-redundantly in the conserved down-regulation pathway to mediate precise endocytic control. Furthermore, they show that this hierarchical mechanism discriminates the endocytic regulation of naturally occurring MOR isoforms. Moreover, they are the first to reveal, we believe, for any seven-transmembrane receptor, a functional role of ubiquitination in the first cytoplasmic loop.     

Does the 'mystery of the extra glucose' during CNS activation reflect glutamate synthesis?

The hypothesis is proposed that the repeatedly demonstrated rise in local cerebral blood flow and glucose utilisation during neuronal activation, without a corresponding increase in oxygen utilisation, may reflect glutamine formation from glucose, followed by complete oxidative degradation of glutamate along complex and extended, but well described pathways, known to operate in the brain. The former process requires large amounts of glucose but little oxygen. The latter utilises oxygen but no additional glucose, is a prolonged process and so at any one time involves only a small increase in the rate of oxygen utilisation which is difficult to demonstrate experimentally.     

Is progesterone a pre-hormone in the CNS?

In this paper, experimental evidences have been presented indicating that progesterone per se appears to be a powerful modulatory steroid of presynaptic striatal dopaminergic terminals of the central nervous system of the rat. This effect of the progesterone signal is concentration as well as infusion mode dependent. Low pulsatile doses of the steroid positively modulate the mechanism by which dopamine terminals respond to amphetamine stimulation and increase tissue dopamine concentration. Whereas, continuous and/or high doses of this steroid negatively modulate the response of the dopamine terminals to amphetamine stimulation and decreases tissue dopamine concentration. This effects occurs through a membrane mediated mechanism either upon the dopamine neuron directly and/or upon an interneuron. Pregnanolone a 5- beta-3 beta-metabolite of progesterone known to activate the hypothalamic LHRH neural apparatus at the level of the hypothalamus of ovariectomized estrogen primed rats in both in vitro as well as in vivo preparations was completely ineffective at the level of the corpus striatum of similar animal preparations. Therefore, it is reasonable to assume that site specific mechanisms exist within the central nervous system which may control differentially the final action of progesterone. In the hypothalamus, pregnanolone appears to be the final signal for its action on the LHRH neural apparatus, whereas in the corpus striatum, the steroid per se, and dependent on the modality and/or the strength of the signal can either directly or indirectly up-regulate (stimulatory component) or down-regulate (inhibitory component) the activity of striatal dopaminergic terminals.     

Does the energy state of mitochondria influence the surface potential of the inner mitochondrial membrane? A critical appraisal

Binding of 8-anilino-1-naphthalene sulphonate (ANS) to rat liver mitochondria and submitochondrial inside-out particles was measured under energized and de-energized conditions. In mitochondria, energization/de-energization changed the binding capacity for ANS extrapolated for its infinitely high concentration, whereas the apparent Kd value remained unchanged. In submitochondrial particles apparent Kd was changed but the extrapolated maximum binding was not altered. These results are compatible with theoretical considerations assuming a free permeability of mitochondrial membranes to ANS and its distribution according to the transmembrane potential. The spin-labelled cationic amphiphile, 4-(dodecyl dimethyl ammonium)-1-oxyl-2,2,6,6-tetramethyl piperidine bromide (CAT12), was trapped by de-energized mitochondria in such a way that about half of the bound probe became inaccessible to reduction by externally added ascorbate. This inaccessible fraction was increased by energization. This indicates that this cationic probe can penetrate through the inner mitochondrial membrane. De-energization produced a parallel shift of the Lineweaver-Burk plots for the oxidation of external ferrocytochrome c by mitoplasts and of succinate by submitochondrial particles. A similar shift was obtained by a partial inhibition of succinate oxidation by antimycin A. Thus, the observed changes of the kinetics of the two membrane-bound enzyme systems on de-energization can be interpreted as reflecting changes of the control points of mitochondrial respiration rather than changes of the surface potential. It is concluded that neither the fluorescent probe ANS, the spin-labelled amphiphilic cation CAT12, nor the kinetics of some respiratory enzyme systems provide a sufficient proof for changes of the surface potential of the inner mitochondrial membrane upon energization.     

Does the margin of stability measure predict medio-lateral stability of gait with a constrained-width base of support?

This study aimed to determine the validity of the centre of mass position (COM) position and extrapolated COM (XCOM), relative to the base of support, for predicting medio-lateral stability during a walking task where the base of support width is limited. Nine young healthy participants walked on a narrow beam. Three-dimensional motion capture was used to calculate the COM and XCOM relative to the base of support. Steps were classified as having either the COM or XCOM inside or outside the base of support, and were classified as successful (stable – foot placed on the beam) or failed (unstable – foot stepped off the beam). If the COM or XCOM are valid measures of stability, they should be within the base of support for successful steps and outside the base of support for failed steps. Classifying the COM and XCOM inside or outside the base of support correctly predicted successful or failed steps in 69% and 58% of cases, respectively. When the COM or XCOM were outside the base of support, walking faster seemed to help participants to maintain stability. The further the COM or XCOM were outside the base of support during a successful step, the more likely participants were to fail on a subsequent step. The results of this study suggest that both COM and XCOM are valid measures of stability during a beam walking task, but that classifying COM and XCOM as inside or outside the base of support may be over-simplistic.     

Does hippocampal size correlate with the degree of caching specialization?

A correlation between the degree of specialization for food hoarding and the volume of the hippocampal formation in passerine birds has been accepted for over a decade. The relationship was first demonstrated in family-level comparisons, and subsequently in species comparisons within two families containing a large number of hoarding species, the Corvidae and the Paridae. Recently, this approach has been criticized as invalid and excessively adaptationist. A recent test of the predicted trends with data pooled from previous studies found no evidence for such a correlation in either of these two families. This result has been interpreted as support for the critique. Here we reanalyse the original dataset and also include additional new data on several parid species. Our results show a surprising difference between continents, with North American species possessing significantly smaller hippocampi than Eurasian ones. Controlling for the continent effect makes the hoarding capacity/hippocampal formation correlation clearly significant in both families. We discuss possible reasons for the continent effect.     

Does the immune system of a mouse age faster than the immune system of a human?

One of the characteristics of all somatic cells is a finite life span. Cells may proliferate until they reach a point after which, although they are metabolically active, they can no longer produce daughter cells. This observation is central to the clonal exhaustion hypothesis, a mechanism cited to explain age-associated immune dysfunction. In this hypothesis, repeated division of lymphocytes leads to a replicative limit, after which they enter the senescent phase but are not lost from the pool of T cells. Advancing age would then be associated with an increase in the number of T cells that are unable to proliferate to a stimulus which induces a proliferative response in T cells from younger individuals. This hypothesis seems both logical and reasonable and is supported by data from both humans and mice with the demonstration of an age-related accumulation of senescent T cells in both species. However, there is an apparent paradox. The paradox arises because the onset of immunosenescence appears to be more closely linked to the life span of the animal rather than the life span of the lymphocyte. BioEssays 21:519–524, 1999. © 1999 John Wiley & Sons, Inc.     

Solution structure of a novel α-conotoxin with a distinctive loop spacing pattern

α-Pharmacological conotoxins are among the most selective ligands of nicotinic acetylcholine receptors with typical cysteine frameworks. They are characterized by the intercysteine loop and classified into various subfamilies, such as α3/5 and α4/7 conotoxins. A novel α-conotoxin, Pu14a (DCPPHPVPGMHKCVCLKTC), with a distinct loop spacing pattern between cysteines was reported recently. Pu14a belongs to the Cys framework 14 (-C-C-C-C) family containing four proline residues in the loop 1 region. Similar to another framework 14 conotoxin Lt14a (MCPPLCKPSCTNC-NH2), Pu14a has C1-C3/C2-C4 disulfide linkage, and can inhibit some subtypes of nicotinic acetylcholine receptors. In this study, the solution structure of Pu14a was investigated using 1H nuclear magnetic resonance spectroscopy to understand the structure-activity relationship of this conotoxin. 20 converged structures of this conopeptide, with RMSD value of 0.77 ?, were obtained based on distance constraints, dihedral angles and disulfide bond constraints. The three-dimensional structure of Pu14a showed remarkable difference from typical α-conotoxins because of a large intercysteine loop between C2 and C13, as well as a 3(10)-helix near the C-terminal. Furthermore, four proline residues in Pu14a adopted the trans conformation that may correlate with the large loop configuration and the biological activity of this conopeptide. The distinct structural characteristics of Pu14a will be very useful for studying the structure-activity relationship of α-conotoxins.     

Information processing in the CNS: a supramolecular chemistry?

How does central nervous system process information? Current theories are based on two tenets: (a) information is transmitted by action potentials, the language by which neurons communicate with each other—and (b) homogeneous neuronal assemblies of cortical circuits operate on these neuronal messages where the operations are characterized by the intrinsic connectivity among neuronal populations. In this view, the size and time course of any spike is stereotypic and the information is restricted to the temporal sequence of the spikes; namely, the “neural code”. However, an increasing amount of novel data point towards an alternative hypothesis: (a) the role of neural code in information processing is overemphasized. Instead of simply passing messages, action potentials play a role in dynamic coordination at multiple spatial and temporal scales, establishing network interactions across several levels of a hierarchical modular architecture, modulating and regulating the propagation of neuronal messages. (b) Information is processed at all levels of neuronal infrastructure from macromolecules to population dynamics. For example, intra-neuronal (changes in protein conformation, concentration and synthesis) and extra-neuronal factors (extracellular proteolysis, substrate patterning, myelin plasticity, microbes, metabolic status) can have a profound effect on neuronal computations. This means molecular message passing may have cognitive connotations. This essay introduces the concept of “supramolecular chemistry”, involving the storage of information at the molecular level and its retrieval, transfer and processing at the supramolecular level, through transitory non-covalent molecular processes that are self-organized, self-assembled and dynamic. Finally, we note that the cortex comprises extremely heterogeneous cells, with distinct regional variations, macromolecular assembly, receptor repertoire and intrinsic microcircuitry. This suggests that every neuron (or group of neurons) embodies different molecular information that hands an operational effect on neuronal computation.     

Does a biological clock reside in the eye of quail?

The site (intra- vs. extraocular) of the circadian clock driving an ocular melatonin rhythm in Japanese quail was investigated by alternately covering the left and right eyes of individual quail, otherwise held in constant light (LL), for 12-hr periods. This procedure exposed each eye to a light-dark (LD) 12:12 light cycle 180 degrees (12 hr) out of phase with the LD 12:12 light cycle experienced by the other eye. This protocol entrained the melatonin rhythm in the left eye of quail 180 degrees out of phase with the rhythm expressed in the right eye. These results are compatible with the hypothesis that an independent light-entrainable circadian pacemaker resides in each eye; they are incompatible with the hypothesis that a single (or functionally single) extraocular pacemaker drives the ocular melatonin rhythm in both eyes. However, the results are also compatible with a model in which two independent extraocular circadian pacemakers, each with an exclusive photic input from one eye, drive the ocular melatonin rhythm.     

Does the growth temperature of a prokaryote influence the purine content of its mRNAs?

The formation and breaking of hydrogen bonds between nucleic acid bases are dependent on temperature. The high G+C content of organisms was surmised to be an adaptation for high temperature survival because of the thermal stability of G:C pairs. However, a survey of genomic GC% and optimum growth temperature (OGT) of several prokaryotes revoked any direct relation between them. Significantly high purine (R=A or G) content in mRNAs is also seen as a selective response for survival among thermophiles. Nevertheless, the biological relevance of thermophiles loading their unstable mRNAs with excess purines (purine-loading or R-loading) is not persuasive. Here, we analysed the mRNA sequences from the genomes of 168 prokaryotes (as obtained from NCBI Genome database) with their OGTs ranging from -5 °C to 100 °C to verify the relation between R-loading and OGT. Our analysis fails to demonstrate any correlation between R-loading of the mRNA pool and OGT of a prokaryote. The percentage of purine-loaded mRNAs in prokaryotes is found to be in a rough negative correlation with the genomic GC% (r(2)=0.655, slope=-1.478, P<000.1). We conclude that genomic GC% and bias against certain combinations of nucleotides drive the mRNA-synonymous (sense) strands of DNA towards variations in R-loading.     

Do light-induced changes in the membrane potential ofNitella reflect the feed-back regulation of a cytoplasmic parameter?

Summary Kinetic studies of the linearized response of membrane potential inNitella to light have revealed the existence of a feedback loop in the pathway of light action. Its existence can hardly be seen in the time course of the responses to dark/light transitions. However, making use of sine waves as input signals and employing a computeraided evaluation has resulted in finding complex time constants in the transfer function of the light effect which point out the existence of a feedback loop. Besides, sometimes spontaneous oscillations with periods of about 1 hr have been observed. It is shown that this system is different from that one reported in literature to be related to cytoplasmic streaming. By measuring the electrical low-frequency impedance, it has been demonstrated that it is not the purpose of the system to control membrane potential, even though secondary effects of the injected current have been found. It seems to be reasonable to assume that this system is involved in the control of a biochemically relevant parameter like the cytoplasmic pH by means of adjusting the balance of counteracting transmembrane transport processes.     

Does interspecific competition with a stronger competitor explain the rarity of an endangered snake on a Mediterranean island?

Interspecific competition has been demonstrated to be an important shaping force for snake communities worldwide, but relatively few studies have investigated its occurrence and extent with island assemblages of snakes. In Sardinia (Mediterranean Sea), two species of whip snakes (Colubridae) co-occur, one of them being abundant and widespread (Hierophis viridiflavus) and the other being localized and critically endangered (Hemorrhois hippocrepis). A previous hypothesis suggested that the latter species would be confined to suboptimal habitats by the former species, which is a stronger competitor for food. As a consequence, He. hippocrepis would not only be rare but would also be smaller in body size in Sardinia than in other regions of its range where Hi. viridiflavus does not occur. In 1999?C2010, we studied habitat selection, food habits, and body sizes of these two snakes in sympatric populations, applying a suite of statistical tools including null models and Monte Carlo simulations. We observed that dietary habits were different between species (compared to Hi. viridiflavus, He. hippocrepis more frequently preyed upon rodents and less frequently on lizards, and within rodent prey, more frequently upon Rattus and less frequently upon Mus), but not in a way compatible with competitively induced trophic niche partitioning. The two species were similar in terms of prey size and predator-size?Cprey-size relationships. They also differed in habitat selection (maquis was the preferred habitat for both, but Hi. viridiflavus was found significantly more often than He. hippocrepis in grasslands, cultivations, and artificial pinewoods) but again in a non-competitively directed way. In addition, there was no evidence for the hypothesis that He. hippocrepis was confined to suboptimal habitats. Body sizes were similar between species (with males being significantly larger than females), and Sardinian He. hippocrepis were not smaller than conspecifics inhabiting regions without Hi. viridiflavus. Overall, our study showed that the two species differed in some aspects of their ecology, but these differences could not be due to competitive interactions. The conservation implications of these results are also discussed.