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Selective neurotoxic damage to the hippocampal formation impairs performance of the transverse patterning and location memory tasks in rhesus macaques.

Monkeys with neurotoxic (ibotenic acid) damage to the hippocampal formation and unoperated controls were trained on two sets of transverse patterning problems (A+/B-, B+/C-, C+/A-, and D+/E-, E+/F-, F+/D-) and a delayed nonmatching-to-location paradigm (DNML) with delays of 10s, 30s, 120s, and 600s. Hippocampal lesions produced a size- and area-dependent impairment on transverse patterning. Damage largely limited to the right hippocampus in one subject had no effect on performance on the task. Of the remaining four subjects, two with hippocampal damage greater than 40% bilaterally were unable to solve the two transverse patterning sets, but could solve the linear set of discriminations (A+/B-, B+/C-, C+/X-). The two remaining operated animals were impaired in acquisition of both sets, but were eventually able to solve one of the two transverse patterning discrimination sets. All five operated monkeys were impaired relative to normal controls on DNML, but not on the standard delayed nonmatching-to-sample (DNMS) version with trial-unique objects. The results confirm our previous findings (Alvarado et al., Hippocampus 12:421-433, 2002) using aspiration lesions of the hippocampal formation and strengthen the view that the hippocampal formation is critical for object and spatial relational memory.

Animals↗

Direct imaging and mesoscale modelling of phase transitions in a nanostructured fluid.

The kinetics of phase transitions is essential for understanding pattern formation in structured fluids. These fluids play a key role in the morphogenesis of biological cells, and they are very common in pharmaceutical products and plastic materials. Until now, it has not been possible to follow phase transitions in structured fluids experimentally in real time and with high spatial resolution. Previous work has relied on static images and indirect experimental evidence from spatially averaging scattering experiments. Simulating the processes with computer models is a further challenge because of the multiple time and length scales involved. Our movies based on in situ scanning force microscopy show the time sequence of the elementary steps of a phase transition in a fluid film of block copolymer from the cylinder to the perforated lamella phase. The movies validate a versatile simulation model that gives physical insight into the nature of the process. Our approach provides a means of improving the study and understanding of pattern formation processes in nanostructured fluids. We expect a significant impact on nanotechnology where block copolymers serve as self-organized templates for the synthesis of inorganic nanostructured materials.

Computer Simulation↗

Spatially restricted patterning cues provided by heparin-binding VEGF-A control blood vessel branching morphogenesis.

Branching morphogenesis in the mammalian lung and Drosophila trachea relies on the precise localization of secreted modulators of epithelial growth to select branch sites and direct branch elongation, but the intercellular signals that control blood vessel branching have not been previously identified. We found that VEGF(120/120) mouse embryos, engineered to express solely an isoform of VEGF-A that lacks heparin-binding, and therefore extracellular matrix interaction domains, exhibited a specific decrease in capillary branch formation. This defect was not caused by isoform-specific differences in stimulating endothelial cell proliferation or by impaired isoform-specific signaling through the Nrp1 receptor. Rather, changes in the extracellular localization of VEGF-A in heparin-binding mutant embryos resulted in an altered distribution of endothelial cells within the growing vasculature. Instead of being recruited into additional branches, nascent endothelial cells were preferentially integrated within existing vessels to increase lumen caliber. The disruption of the normal VEGF-A concentration gradient also impaired the directed extension of endothelial cell filopodia, suggesting that heparin-binding VEGF-A isoforms normally provide spatially restricted stimulatory cues that polarize and thereby guide sprouting endothelial cells to initiate vascular branch formation. Consistent with this idea, we found opposing defects in embryos harboring only a heparin-binding isoform of VEGF-A, including excess endothelial filopodia and abnormally thin vessel branches in ectopic sites. We conclude that differential VEGF-A isoform localization in the extracellular space provides a control point for regulating vascular branching pattern.

Animals↗

Regionalization and segmentation of the leech.

Regionalization and segmentation of the leech body plan have been examined by numerous approaches over the years. A wealth of knowledge has accumulated regarding the normally invariant cell lineages of the leech and the degree of developmental plasticity that is possible in each cell line in early development and in neurogenesis. Homologues of genes that control regionalization and segmentation in Drosophila have been cloned from the leech and the expression patterns reveal conserved features with those in Drosophila and other organisms. Possible developmental functions of the en-class proteins in spatial and temporal modes of segment formation are discussed in light of leech and Drosophila development. Annelida and Arthropoda cell lineages of engrailed-class gene expression are compared in leech blast cell clones and crustacean parasegments. In addition, future directions for molecular analysis of segmentation of the leech are summarized.

Animals↗

Equivalent spatial location memory deficits in rats with medial septum or hippocampal formation lesions and patients with dementia of the Alzheimer's type.

College students, healthy elderly subjects, patients diagnosed with mild or moderate dementia of the Alzheimer's type, as well as rats with small or large lesions of the medial septum (MS), dorsal hippocampal formation (DHF) or nucleus basalis magnocellularis (NBM) were tested on an item memory task for a five- or six-item list of varying spatial locations. Equivalent patterns of item memory deficits as a function of serial order position were observed in rats with small or large MS or DHF lesions and patients with mild or moderate dementia of the Alzheimer's type. No deficits were found for NBM-lesioned rats. The results provide support for the possibility that rats with MS and DHF lesions mimic the mnemonic symptomatology of patients with Alzheimer's disease.

Acetylcholinesterase↗

Establishment of dorsal-ventral and terminal pattern in the Drosophila embryo.

Dorsal-ventral and terminal pattern formation in the Drosophila embryo is mediated via inductive signals originating during oogenesis from the somatic follicle cells that ensheath the developing oocyte. This somatically expressed spatial information controls dorsal-ventral development by defining the polarity of a signal transduction pathway that results in the graded nuclear concentration of the dorsal gene product, a transcription factor.

Animals↗

Structural characterization of folding intermediates in cytochrome c by H-exchange labelling and proton NMR.

To understand the process of protein folding, it will be necessary to obtain detailed structural information on folding intermediates. This difficult problem is being studied by using hydrogen exchange and rapid mixing to label transient structural intermediates, with subsequent analysis of the proton-labelling pattern by two-dimensional nuclear magnetic resonance spectroscopy. Results for cytochrome c show that the method provides the spatial and temporal resolution necessary to monitor structure formation at many defined sites along the polypeptide chain on a timescale ranging from milliseconds to minutes.

Cytochrome c Group↗

Spatio-temporal nonlinear modeling of gastric myoelectrical activity.

The accomplishment of a complete digestive process of human stomach is regulated by a spatio-temporally-coordinated electric pattern called gastric myoelectrical activity (GMA). The normal patterns of GMA present temporal evolution from endogenous rhythmic oscillation to bursting of spikes associated with contractions, and also ordered spatial propagation of the oscillating waves. The abnormal patterns of GMA have been observed in temporal dysrhythmia, such as tachygastria, bradygastria and arrhythmia, and in spatial propagation failure, such as retrograde propagation and uncoupling. Different GMA patterns are associated with different gastric symptoms and there exist some nonlinear mechanisms to govern the formation and dynamic evolution of these patterns. However, these mechanisms are so complex that few of them are known by medical observations. The aim of this study is to explore these mechanisms by spatio-temporal modeling of GMA. The single-cell model simulating the formation process of slow waves and spikes, the multi-cell model simulating the propagation process of GMA and the extracellular model simulating the formation of bipolar recordings are presented.

Computer Simulation↗

Ultrasound and electromagnetic radiation in hyperthermia--a historical perspective.

An account is given of the historical developments of ultrashortwave, microwave and ultrasonic diathermy techniques, for which bio-physical and bio-engineering principles were developed mainly after the Second World War. Topics included in this survey are: electric and acoustic properties of tissues, penetration depth and focusing ability for ultrasound (US) and electro-magnetic (EM) radiations, heat deposition patterns in complex tissue layers, hot-spot formation, temperature elevations in local areas of elevated heat deposition (SAR), and thermoregulatory responses. The present possibilities of placing spatially defined increases in temperature in tissue by either ultrasound or radio frequency radiation are summarized.

Diathermy↗

Expression of pattern in plants: combining molecular and calculus-based biophysical paradigms.

Pattern formation in plant meristems occurs across a broad scale. At the topographical level (large scale), tissue folding in the meristem is responsible for the initiation of new organs in specific phyllotactic patterns and also determines organ shape. At the cellular level (small scale), oriented cell division and microtubule-based cellulose reinforcement control cell pattern and growth direction. I argue here that structural specification at each scale is highly efficient if the pertinent gene activity is manifested in two complementary biophysical categories. At large scale, one category is the tendency of the formative tissue to fold with a certain spatial periodicity determined by its material properties (e.g., bending stiffness from cellulose content). This latent tendency is formalized in a differential equation for physical buckling. The second category at this scale comprises boundary conditions that specify how the latent tendency is manifested as topography: whether tissue humps occur as whorls or Fibonacci spirals. This versatile combinatorial format accounts for the relative stability of alternative organ patterning as well as alternative organ shaping (e.g., stamens vs. carpels). It also accounts for the structural shifts seen in normal development and after mutation or chemical/physical intervention. At small scale, the latent differential activity is the tendency for groups of dividing cells to co-align their cytoskeletons. The curvature of the surface opposes this tendency. The least curved part of a new primordium is its quasicylindrical midportion. There, by aligning microtubules and cellulose coherently around the organ, a new growth direction is set. Thus large-scale buckling produces curvature variation, which, in turn, affects the localization and orientation of the cytoskeleton. This scheme for the coherent production of diverse geometrical features, involving calculus at two structural levels, is supported by complex organogenetic responses to simple physical intervention. Also, many morphological alternatives, wild type vs. mutant, reflect single changes in parameters in this differential-integral format.

Journal Article↗

CaMKII activation in the entorhinal cortex disrupts previously encoded spatial memory.

To investigate the role of the entorhinal cortex in memory at a molecular level, we developed transgenic mice in which transgene expression was inducible and limited to the superficial layers of the medial entorhinal cortex, pre- and parasubiculum. We found that expression of a constitutively active mutant form of CaMKII in these structures disrupted spatial memory formation. Immediate post-training activation of the transgene disrupted previously established memory while transgene activation 3 weeks following the training was ineffective. These results demonstrate that, similar to the hippocampus, the entorhinal cortex plays a time-limited role in spatial memory formation but is not a final cortical repository of long-term memory. Moreover, these results suggest that the indiscriminate activation of CaMKII is able to disrupt preexisting memories, possibly by altering the pattern of synaptic weight changes that are thought to form the basis of the memory trace.

Animals↗

Normal ontogenic observations on the expression of Eph receptor tyrosine kinase, Cek8, in chick embryos.

The spatio-temporal pattern of expression for the Eph receptor tyrosine kinase, Cek8, was observed in normal chick embryos from H-H stage 6 to 23 by immunohistochemical techniques. Expression of Cek8 was already present in embryos at H-H stage 6, where it was located in the neural plate of the brain region, paraxial mesoderm, and the primitive streak. Regions expressing Cek8 subsequently increased during development to include the neural folds of the brain, rhombomeres 3 and 5, the caudal part of the neural plate, neural crest cells related to the formation of glossopharyngeal nerve ganglia, invaginated cells throughout the primitive groove and the epithelium of the rudiment of the gall bladder. Cek8 was also expressed in the mesenchymal cells of the pharyngeal arches, allantoic stalk and limb buds as well as in the areas surrounding the eye vesicles and nasal pits. Furthermore, cells in the tail bud progressing to the secondary neurulation expressed Cek8. Thus, the spatiotemporal patterns of Cek8 expression appears to have intimate relationships with tissue rebuilding, the maturation of differentiated cells, and the spatial organization of tissues. Consequently, it appears that Cek8 plays an integral role in the developmental events leading to the formation of a wide--though not inclusive--variety of tissues and organ systems.

Animals↗

Spatial and temporal distribution of sonic hedgehog mRNA in the embryonic mouse mandible by reverse transcription/polymerase chain reaction and in situ hybridization analysis.

Hedgehog genes have recently been implicated in the control of pattern formation in many developing organ system. Vertebrate homologues of the Drosophila hedgehog have been identified in mouse and rate embryos. The temporal regulation of sonic hedgehog (mouse homologue) has previously been studied by Northern analysis of whole embryos with varying results. Sonic hedgehog transcript expression in the mouse mandibular process was now characterized using polymerase chain reaction (PCR) an in situ hybridization techniques. PCR analysis revealed transcripts at gestational days 10 and 11, before the formation of the dental lamina, but not at days 12-14, after tooth buds have formed. Transcripts were localized to, primarily, the epithelium in the presumptive incisor region of the mandibular midline at gestational day 10. No mRNA was detected by in situ hybridization techniques in the presumptive molar regions of odontogenic epithelium. Sonic hedgehog expression may be involved in the regulation of pattern formation through establishment of an incisor-molar axis of polarity.

Actins↗

A role for Fringe in segment morphogenesis but not segment formation in the grasshopper, Schistocerca gregaria.

Studies of somitogenesis in vertebrates have identified a number of genes that are regulated by a periodic oscillator that patterns the pre-somitic mesoderm. One of these genes, hairy, is homologous to a Drosophila segmentation gene that also shows periodic spatial expression. This, and the periodic expression of a zebrafish homologue of hairy during somitogenesis, has suggested that insect segmentation and vertebrate somitogenesis may use similar molecular mechanisms and possibly share a common origin. In chicks and mice expression of the lunatic fringe gene also oscillates in the presomitic mesoderm. Fringe encodes an extracellular protein that regulates Notch signalling. This, and the finding that mutations in Notch or its ligands disrupt somite patterning, suggests that Notch signalling plays an important role in vertebrate somitogenesis. Although Notch signalling is not known to play a role in the formation of segments in Drosophila, we reasoned that it might do so in other insects such as the grasshopper, where segment boundaries form between cells, not between syncytial nuclei as they do in Drosophila. Here we report the cloning of a single fringe gene from the grasshopper Schistocerca. We show that it is not detectably expressed in the forming trunk segments of the embryo until after segment boundaries have formed. We conclude that fringe is not part of the mechanism that makes segments in Schistocerca. Thereafter it is expressed in a pattern which shows that it is a downstream target of the segmentation machinery and suggests that it may play a role in segment morphogenesis. Like its Drosophila counterpart, Schistocerca fringe is also expressed in the eye, in rings in the legs, and during oogenesis, in follicle cells.

Amino Acid Sequence↗

To shape a cell: an inquiry into the causes of morphogenesis of microorganisms.

We recognize organisms first and foremost by their forms, but how they grow and shape themselves still largely passes understanding. The objective of this article is to survey what has been learned of morphogenesis of walled eucaryotic microorganisms as a set of problems in cellular heredity, biochemistry, physiology, and organization. Despite the diversity of microbial forms and habits, some common principles can be discerned. (i) That the form of each organism represents the expression of a genetic program is almost universally taken for granted. However, reflection on the findings with morphologically aberrant mutants suggests that the metaphor of a genetic program is misleading. Cellular form is generated by a web of interacting chemical and physical processes, whose every strand is woven of multiple gene products. The relationship between genes and form is indirect and cumulative; therefore, morphogenesis must be addressed as a problem not of molecular genetics but of cellular physiology. (ii) The shape of walled cells is determined by the manner in which the wall is laid down during growth and development. Turgor pressure commonly, perhaps always, supplies the driving force for surface enlargement. Cells yield to this scalar force by localized, controlled wall synthesis; their forms represent variations on the theme of local compliance with global force. (iii) Growth and division in bacteria display most immediately the interplay of hydrostatic pressure, localized wall synthesis, and structural constraints. Koch's surface stress theory provides a comprehensive and quantitative framework for understanding bacterial shapes. (iv) In the larger and more versatile eucaryotic cells, expansion is mediated by the secretion of vesicles. Secretion and ancillary processes, such as cytoplasmic transport, are spatially organized on the micrometer scale. The diversity of vectorial physiology and of the forms it generates is illustrated by examples: apical growth of fungal hyphae, bud formation in yeasts, germination of fucoid zygotes, and development of cells of Nitella, Closterium, and other unicellular algae. (v) Unicellular organisms, no less than embryos, have a remarkable capacity to impose spatial order upon themselves with or without the help of directional cues. Self-organization is reviewed here from two perspectives: the theoretical exploration of morphogens, gradients, and fields, and experimental study of polarization in Fucus cells, extension of hyphal tips, and pattern formation in ciliates. Here is the heart of the matter, yet self-organization remains nearly as mysterious as it was a century ago, a subject in search of a paradigm.

Animals↗

Heterocyst differentiation and pattern formation in cyanobacteria: a chorus of signals.

Heterocyst differentiation in filamentous cyanobacteria provides an excellent prokaryotic model for studying multicellular behaviour and pattern formation. In Anabaena sp. strain PCC 7120, for example, 5-10% of the cells along each filament are induced, when deprived of combined nitrogen, to differentiate into heterocysts. Heterocysts are specialized in the fixation of N(2) under oxic conditions and are semi-regularly spaced among vegetative cells. This developmental programme leads to spatial separation of oxygen-sensitive nitrogen fixation (by heterocysts) and oxygen-producing photosynthesis (by vegetative cells). The interdependence between these two cell types ensures filament growth under conditions of combined-nitrogen limitation. Multiple signals have recently been identified as necessary for the initiation of heterocyst differentiation, the formation of the heterocyst pattern and pattern maintenance. The Krebs cycle metabolite 2-oxoglutarate (2-OG) serves as a signal of nitrogen deprivation. Accumulation of a non-metabolizable analogue of 2-OG triggers the complex developmental process of heterocyst differentiation. Once heterocyst development has been initiated, interactions among the various components involved in heterocyst differentiation determine the developmental fate of each cell. The free calcium concentration is crucial to heterocyst differentiation. Lateral diffusion of the PatS peptide or a derivative of it from a developing cell may inhibit the differentiation of neighbouring cells. HetR, a protease showing DNA-binding activity, is crucial to heterocyst differentiation and appears to be the central processor of various early signals involved in the developmental process. How the various signalling pathways are integrated and used to control heterocyst differentiation processes is a challenging question that still remains to be elucidated.

Anabaena↗

Neuronal expression and interaction with the synaptic protein CASK suggest a role for Neph1 and Neph2 in synaptogenesis.

Formation, differentiation, and plasticity of synapses require interactions between pre- and postsynaptic partners. Recently, it was shown that the transmembrane immunoglobulin superfamily protein SYG-1 is required for providing synaptic specificity in C. elegans. However, it is unclear whether the mammalian orthologs of SYG-1 are also involved in local cell interactions to determine specificity during synapse formation. We used in situ hybridization, immunohistochemistry, and immunogold electron microscopy to study the temporal and spatial expression of Neph1 and Neph2 in the developing and adult mouse brain. Both proteins show similar patterns with neuronal expression starting around embryonic days 12 and 11, respectively. Expression is strongest in areas of high migratory activity. In the adult brain, Neph1 and Neph2 are predominantly seen in the olfactory nerve layer and the glomerular layer of the olfactory bulb, in the hippocampus, and in Purkinje cells of the cerebellum. At the ultrastructural level, Neph1 and Neph2 are detectable within the dendritic shafts of pyramidal neurons. To a lesser extent, there is also synaptic localization of Neph1 within the stratum pyramidale of the hippocampal CA1 and CA3 region on both pre- and postsynaptic sites. Here it colocalizes with the synaptic scaffolder calmodulin-associated serin/threonin kinase (CASK), and both Neph1 and Neph2 interact with the PDZ domain of CASK via their cytoplasmic tail. Our results show that Neph proteins are expressed in the developing nervous system of mammals and suggest that these proteins may have a conserved function in synapse formation or neurogenesis.

Animals↗

Diffusion resistances between ADH-induced vacuoles and the extracellular space in rabbit collecting duct: evidence that most vacuoles are intracellular, endocytic compartments.

Large vacuoles form in the renal collecting duct following the onset of antidiuretic hormone (ADH)-stimulated water reabsorption. The aim of the present study was to test two alternative hypotheses regarding the origins of these structures: (1) the vacuoles constitute basilar, extracellular spaces that dilate as water flows through these spaces from cells into the peritubular compartment; or (2) the vacuoles represent intracellular, endocytic compartments that dilate during water reabsorption due to enhanced fluid phase endocytosis. Fluorescence-digital imaging microscopy was used to visualize the uptake into vacuoles of a hydrophilic fluorochrome (6 methoxy-N-[3 sulfopropyl] quinolinium) whose fluorescence is markedly quenched by halides. During their formation, most vacuoles (67%) accumulated the fluorochrome from the peritubular bath and trapped the dye well after (greater than 60 min) washing it from the bath. The spatial pattern of fluorescence within individual vacuoles indicated that the dye was trapped within these structures as a fluid-phase marker and was not bound to the vacuole margins. The fluorescence of dye trapped within vacuoles was virtually unaltered by changes in peritubular Cl- or Br- concentration that elicit dramatic quenching of dye-fluorescence in bulk solution, as expected if there exists a high diffusion resistance between the interiors of these structures and the peritubular space. These results indicate that most ADH-induced vacuoles represent endocytic compartments that are not directly connected to the extracellular space.

Animals↗