Morphological and biomechanical studies of the ligamentum arteriosum

Fifteen ligaments taken from individuals aged between 60 and 80 years were used for the study of the histological structure, the composition of the ground substance and the biomechanical behavior. Remnants of the original duct are recognizable in the ligament as artery of the muscular type. What had been the intima is thickened and consists mainly of cell-poor, fiber-rich connective tissue, which often shows chondrification along with calcification. The biomechanical behavior of the ligaments, evaluated with the force-length and force-relaxation test, was similar to that of peripheral arteries.     

Functional architecture of the CFTR chloride channel

Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a member of the ATP-binding cassette (ABC) family of membrane transport proteins. CFTR is unique among ABC proteins in that it functions not as an active transporter but as an ATP-gated Cl^? channel. As an ion channel, the function of the CFTR transmembrane channel pore that mediates Cl^? movement has been studied in great detail. On the other hand, only low resolution structural data is available on the transmembrane parts of the protein. The structure of the channel pore has, however, been modeled on the known structure of active transporter ABC proteins. Currently, significant barriers exist to building a unified view of CFTR pore structure and function. Reconciling functional data on the channel with indirect structural data based on other proteins with very different transport functions and substrates has proven problematic. This review summarizes current structural and functional models of the CFTR Cl^? channel pore, including a comprehensive review of previous electrophysiological investigations of channel structure and function. In addition, functional data on the three-dimensional arrangement of pore-lining helices, as well as contemporary hypotheses concerning conformational changes in the pore that occur during channel opening and closing, are discussed. Important similarities and differences between different models of the pore highlight current gaps in our knowledge of CFTR structure and function. In order to fill these gaps, structural and functional models of the membrane-spanning pore need to become better integrated.     

Functional architecture of auditory cortex

Three complementary approaches demonstrate new types of organization in rodent, feline and primate auditory cortex, as well as differences in processing between auditory and visual cortex. First, connectional work reveals patterns of thalamocortical and corticocortical input unique to the auditory cortex. Second, physiological studies find multiple, interleaved auditory processing modules related to corticocortical connections and embedded in the isofrequency gradient. Third, functional analyses demonstrate independent processing streams for sound localization and identification analogous to the 'what' and 'where' streams in visual cortex, although the modular arrangements are modality-specific. Taken together, these data show that the auditory cortex has common and unique functional substrates.     

Functional architecture of the splenic vein in the adult human

The spatial arrangement of the muscular, elastic and collagenous fibers was examined in 20 splenic veins of adult humans. Histologic and panorganographic sections were stained by the following methods: azan, resorcinfuchsin (Weigert), Verhoeff-van Gieson's stain and Masson's trichrome. Unstained sections were examined under polarized light. The results show that the muscular and collagenous fibers of the splenic vein exhibit an elongated spiral arrangement with no polarization. The elastic fibers form a threedimensional network disposed predominantly in a longitudinal direction. Hemodynamics are discussed based on the spatial structure of the muscular, elastic and collagenous elements of the vein wall and its relationship with the circulatory functions of the spleen.     

Functional dissection of the apicomplexan glideosome molecular architecture

The glideosome of apicomplexan parasites is an actin- and myosin-based machine located at the pellicle, between the plasma membrane (PM) and inner membrane complex (IMC), that powers parasite motility, migration, and host cell invasion and egress. It is composed of myosin A, its light chain MLC1, and two gliding-associated proteins, GAP50 and GAP45. We identify GAP40, a polytopic protein of the IMC, as an additional glideosome component and show that GAP45 is anchored to the PM and IMC via its N- and C-terminal extremities, respectively. While the C-terminal region of GAP45 recruits MLC1-MyoA to the IMC, the N-terminal acylation and coiled-coil domain preserve pellicle integrity during invasion. GAP45 is essential for gliding, invasion, and egress. The orthologous Plasmodium falciparum GAP45 can fulfill this dual function, as shown by transgenera complementation, whereas the coccidian GAP45 homolog (designated here as) GAP70 specifically recruits the glideosome to the apical cap of the parasite.     

Functional architecture of long-range perceptual interactions

The pattern of lateral interactions in the primary visual cortex, which has emerged from recent studies, conforms to the grouping rules of similarity, proximity, smoothness and closure. The goal of this paper is to understand the perceptual salience of oriented elements that are specifically organized to form a smooth contour. An overview of recent studies, in combination with new experimental results, is presented here to emphasis the idea that visual responses depend on input from both the center and the surround of the classical receptive field (CRF). It is assumed that normal lateral interactions produce a neuronal network that is formed by two antagonistic mechanisms: (i) excitation, that is spatially organized along the optimal orientation (collinear), and is predominant near the contrast threshold of the neuron, and (ii) inhibition, that is less selective and is distributed diffusely around the cell's response field. Thus, the inputs from the CRF and the anisotropic surround are summated non-linearly. The specificity of the facilitation and suppression along the collinear direction suggests the existence of second-order elongated collinear filters, which may increase the response similarity between neurons responding to elongated stimulus, thus may enhance the perceptual salience of anisotropic configurations such as contours. This causal connection is particularly evident in amblyopes, where abnormal development of the network results in the abnormal perception of contours.     

Functional architecture of RNA polymerase I

Synthesis of ribosomal RNA (rRNA) by RNA polymerase (Pol) I is the first step in ribosome biogenesis and a regulatory switch in eukaryotic cell growth. Here we report the 12 A cryo-electron microscopic structure for the complete 14-subunit yeast Pol I, a homology model for the core enzyme, and the crystal structure of the subcomplex A14/43. In the resulting hybrid structure of Pol I, A14/43, the clamp, and the dock domain contribute to a unique surface interacting with promoter-specific initiation factors. The Pol I-specific subunits A49 and A34.5 form a heterodimer near the enzyme funnel that acts as a built-in elongation factor and is related to the Pol II-associated factor TFIIF. In contrast to Pol II, Pol I has a strong intrinsic 3'-RNA cleavage activity, which requires the C-terminal domain of subunit A12.2 and, apparently, enables ribosomal RNA proofreading and 3'-end trimming.     

Functional architecture of the outer arm dynein conformational switch

Dynein light chain 1 (LC1/DNAL1) is one of the most highly conserved components of ciliary axonemal outer arm dyneins, and it associates with both a heavy chain motor unit and tubulin located within the A-tubule of the axonemal outer doublet microtubules. In a variety of model systems, lack of LC1 or expression of mutant forms leads to profound defects in ciliary motility, including the failure of the hydrodynamic coupling needed for ciliary metachronal synchrony, random stalling during the power/recovery stroke transition, an aberrant response to imposed viscous load, and in some cases partial failure of motor assembly. These phenotypes have led to the proposal that LC1 acts as part of a mechanical switch to control motor function in response to alterations in axonemal curvature. Here we have used NMR chemical shift mapping to define the regions perturbed by a series of mutations in the C-terminal domain that yield a range of phenotypic effects on motility. In addition, we have identified the subdomain of LC1 involved in binding microtubules and characterized the consequences of an Asn → Ser alteration within the terminal leucine-rich repeat that in humans causes primary ciliary dyskinesia. Together, these data define a series of functional subdomains within LC1 and allow us to propose a structural model for the organization of the dynein heavy chain-LC1-microtubule ternary complex that is required for the coordinated activity of dynein motors in cilia.     

The architecture of the reflex arc]

The conceptual reflex arc in Helix is composed of identifiable local detectors, command neurons and modulatory neurons. The identification of neurons opens a perspective for identification of single synapses. The plastic changes within the reflex arc can be characterized as non-associative and associative. The non-associative plasticity (habituation and sensitization) is based on the dephosphorylation and phosphorylation of receptor and channel proteins. The associative plasticity (conditioning and extinction) is based on two levels of molecular modifications: the short-term changes underlined by the phosphorylation and dephosphorylation of receptor and channel molecules and the long-term changes referred to the expression of genes encoding structural and translocating proteins. The very selective receptor and channel modifications are due to the Hebbian plastic synapse supplemented with the principle of nonspecific (modulating) influences.     

Functional architecture of the SR calcium store in uterine smooth muscle

Sarcoplasmic reticulum (SR) is abundant in uterine smooth muscle cells. The functional role of this organelle in the regulation of uterine myocytes is not fully understood. The data available in the literature suggest that SR plays a dual role: as a source of calcium and as a calcium sink shaping calcium transients produced by membrane depolarisation and uterotonic agonists. Advances in digital imaging techniques including confocal microscopy of isolated living cells, and the development of methods for direct measurement of intraluminal calcium, has triggered a substantial increase in the number of publications elucidating the role of intracellular stores in calcium signalling. In this paper we review the literature and our own work on the SR calcium store in uterine smooth muscle cells.     

Functional architecture of vasopressin/oxytocin receptors

Three-dimensional models of G protein-coupled receptors (GPCR) have been defined using most experimental data available and protein modeling techniques. The endogenous ligand binding sites have been qualitatively described and putative receptor activation mechanisms have been proposed. The model has been recently refined to take into account recent crystallographic data. Most experimental results published are in excellent qualitative agreement with the initial model. We have undertaken to study more systematically by site directed mutagenesis the vasopressin/oxytocin receptor binding domain as a prototype of neuropeptide receptors. The experimental results are in very good agreement with the models. The residues responsible for the neuropeptide binding have been identified and confirm the predicted localization of the neuromediator in the transmembrane domain of the receptors. The side chain of the 8th residue of vasopressin interacts with a non-conserved receptor residue located in the first extracellular loop. As predicted from the model, this interaction is completely responsible for the selectivity of the ligand-receptor interaction. Finally, aromatic residues which allow the modulation of the efficacy of agonists have been identified.     

Functional architecture of olfactory ionotropic glutamate receptors

Download : Download video (23MB)     

Functional architecture of higher plant photosystem II supercomplexes

Photosystem II (PSII) is a large multiprotein complex, which catalyses water splitting and plastoquinone reduction necessary to transform sunlight into chemical energy. Detailed functional and structural studies of the complex from higher plants have been hampered by the impossibility to purify it to homogeneity. In this work, homogeneous preparations ranging from a newly identified particle composed by a monomeric core and antenna proteins to the largest C₂S₂M₂ supercomplex were isolated. Characterization by biochemical methods and single particle electron microscopy allowed to relate for the first time the supramolecular organization to the protein content. A projection map of C₂S₂M₂ at 12 Å resolution was obtained, which allowed determining the location and the orientation of the antenna proteins. Comparison of the supercomplexes obtained from WT and Lhcb‐deficient plants reveals the importance of the individual subunits for the supramolecular organization. The functional implications of these findings are discussed and allow redefining previous suggestions on PSII energy transfer, assembly, photoinhibition, state transition and non‐photochemical quenching.     

Functional analysis of hairpin ribozyme active site architecture

The hairpin ribozyme is a small catalytic motif found in plant satellite RNAs where it catalyzes a reversible self-cleavage reaction during processing of replication intermediates. Crystallographic studies of hairpin ribozymes have provided high resolution views of the RNA functional groups that comprise the active site and stimulated biochemical studies that probed the contributions of nucleobase functional groups to catalytic chemistry. The dramatic loss of activity that results from perturbation of active site architecture points to the importance of positioning and orientation in catalytic rate acceleration. The current study focuses on the network of noncovalent interactions that align nucleophilic and leaving group oxygens in the orientation required for the S(N)2-type reaction mechanism and orient the active site nucleobases near the reactive phosphate to facilitate catalytic chemistry. Nucleotide modifications that alter or eliminate individual hydrogen bonding partners had different effects on the activation barrier to catalysis, the stability of ribozyme complexes in the ground state, and the internal equilibrium between cleavage and ligation of bound products. Furthermore, substitution of hydrogen bond donors and acceptors with seemingly equivalent pairs sometimes had very different functional consequences. These biochemical analyses augment high resolution structural information to provide insights into the functional significance of active site architecture.     

Functional architecture of neural circuits for leg proprioception in Drosophila

1. Download : Download high-res image (252KB)
2. Download : Download full-size image

     

Functional and clinical significance of the architecture of human skeletal muscles

The architectural properties of the triceps surae muscle were studied in vivo in groups of healthy subjects (eight men) and patients with locomotor function disorders (four men and four women) with the ankle joint positioned at a plantar flexion 0° and the knee set at 90° (neutral position). In this position, using ultrasonic scanning, longitudinal ultrasonic images of the medial gastrocnemius (MG), lateral gastrocnemius (LG), and soleus (Sol) muscles were obtained when the subject was relaxed (the passive state) or performed isometric plantar flexion (50% of the maximum voluntary contraction (MVC), the active state). The fascicle lengths, fascicle angles, and muscle thickness were determined. In the passive state, the fascicle lengths of the MG, LG, and Sol muscles in the group of healthy subjects were 33, 35, and 30 mm and the pennation angle, 25°, 19°, and 25°; in the group of patients with motor disorders, 38, 39, and 29 mm and 21°, 19°, and 24°, respectively. The MG, LG, and Sol thicknesses in the group of healthy subjects were 15, 13, and 12 mm, and in the group of patients with motor disorders, 14, 12, and 14 mm, respectively. In the active state (50% of the MVC), the MG, LG, and Sol fiber lengths in the group of healthy subjects shortened by 31, 24, and 18%; the fiber pennation angle increased by 60, 41, and 41%, respectively. In the group of patients with motor disorders, the fiber lengths shortened by 28, 14, and 18% and the fiber pennation angle decreased by 28, 26, and 36%, respectively. The MG, LG, and Sol thicknesses in the group of healthy subjects increased by 9, 22, and 18%, while in the group of patients with motor disorders the thickness decreased by 4% in the MG and increased by 11 and 4% in the LG and Sol muscles, respectively. Different fiber lengths and pennation angles and their changes upon contraction might be related to differences in the force-producing capabilities of the muscles and the viscoelastic properties of muscle tendons and aponeuroses.