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PubMed2005

Basigin (EMMPRIN/CD147) interacts with integrin to affect cellular architecture.

Basigin, an IgG family glycoprotein found on the surface of human metastatic tumors, stimulates fibroblasts to secrete matrix metalloproteases that remodel the extracellular matrix. Using Drosophila melanogaster we identify intracellular, matrix metalloprotease-independent, roles for basigin. Specifically, we found that basigin, interacting with integrin, is required for normal cell architecture in some cell types. Basigin promotes cytoskeletal rearrangements and the formation of lamellipodia in cultured insect cells. Loss of basigin from photoreceptors leads to misplaced nuclei, rough ER and mitochondria, as well as to swollen axon terminals. These changes in intracellular structure suggest cytoskeletal disruptions. These defects can be rescued by either fly or mouse basigin. Basigin and integrin colocalize to cultured cells and to the visual system. Basigin-mediated changes in the architecture of cultured cells require integrin binding activity. Basigin and integrin interact genetically to affect cell structure in the animal, possibly by forming complexes at cell contacts that help organize internal cell structure.

Actins↗

PubMed2005

Chromosome architecture in the decondensing human sperm nucleus.

Whereas recent studies demonstrated a well-defined nuclear architecture in human sperm nuclei, little is known about the mode of DNA compaction above the elementary structural unit of nucleoprotamine toroids. Here, using fluorescence in-situ hybridization (FISH) with arm-specific DNA probes of chromosomes 1, 2 and 5, we visualized arm domains and established hierarchical levels of sperm chromatin structures. The compact chromosome territories, which in sperm have a preferred intranuclear localization, have an extended conformation represented by a 2000 nm chromatin fiber. This fiber is composed of a 1000 nm chromatin thread bent at 180 degrees near centromere. Two threads of 1000 nm, representing p-arm and q-arm chromatin, run in antiparallel fashion and join at the telomeres. Each 1000 nm thread, in turn, resolves into two rows of chromatin globules 500 nm in diameter interconnected with thinner chromatin strands. We propose a unified comprehensive model of chromosomal and nuclear architecture in human sperm that, as we suggest, is important for successful fertilization and early development.

Cell Nucleus↗

PubMed2007

Transmitting on actin: synaptic control of dendritic architecture.

Excitatory synaptic transmission in the central nervous system mainly takes place at dendritic spines, highly motile protrusions on the dendritic surface. Depending on the stimuli received, dendritic spines undergo rapid actin-based changes in their morphology. This plasticity appears to involve signaling through numerous proteins that control the organization of the actin cytoskeleton (actin regulators). At least in part, recruitment and activation of these depends on neurotransmitter receptors at the post-synapse, which directly link neurotransmission to changes in dendritic spine architecture. However, other, non-neurotransmitter-receptors present at dendritic spines also participate. It is likely that several receptor types can control the activity of a single actin-regulatory pathway and it is the complex integration of numerous signals that determines the overall architecture of a dendritic spine.

Actins↗

PubMed1992

A functional model of adult human prostate epithelium. The role of androgens and stroma in architectural organisation and the maintenance of differentiated secretory function.

A functional model of adult human prostate epithelium is described. This model shows that stromal cells, but not an androgenic stimuli, are required for architectural organisation of prostate epithelium. Within an organised structure, androgenic stimulation is required for the establishment of secretory epithelial cell morphology and associated function. In the absence of stromal cells but in the presence of androgens architectural organisation and secretory function are lost. Epithelial parenchymal units (organoids) from human prostate tissue were isolated, cultured within a three-dimensional collagen matrix, and xenografted subcutaneously into athymic mouse hosts. The grafted gels were rapidly invaded by host fibroblasts. Epithelial organisation initially disappeared but was re-established concurrently with the stromal cell invasion. In intact male hosts, cuboidal and columnar cells that expressed human prostate-specific secretory markers were found. In castrated male and in female hosts epithelial structures were lined with flattened epithelium with no secretory function. This phenomenon could be reversibly replicated by treating intact male hosts with the anti-androgen Flutamide. Gels containing organoids grafted within 0.45 microns Millipore chambers were not invaded by stromal cells and rapidly lost all epithelial organisation and secretory function. When organoids cocultured with human foreskin fibroblasts were grafted within chambers, structural organisation of the epithelium was supported. These results indicate that both heterologous human fibroblasts and mouse stromal cells are capable of permissively supporting adult human prostate epithelial function.

Aged↗

PubMed1995

The three-dimensional organization of ribosomal genes and the architecture of the nucleoli vary with G1, S and G2 phases.

The three-dimensional (3-D) organization of the nucleolus, a defined nuclear territory containing tandem repeats of the ribosomal genes (rDNA), was investigated in PtK1 cells. Identification of the interphase stages was performed in single cells using DNA quantification by cytometry for the G1 and G2 phases while the S phase was identified by immunolabelling of the proliferating cell nuclear antigen (PCNA). The 3-D organization of the rDNA in the nucleolus was analyzed by fluorescence in situ hybridization using confocal microscopy. All the rDNA was located inside the nucleolar structures during all stages and the two rDNA loci were orthogonal. The rDNA was heterogeneously distributed in each nucleolus during G1, S and G2, with alternate sites of clustered genes (spots) and of genes in more extended configurations. The number of spots (4 to 6 in G1) increased during S phase (7 to 12) and their 3-D organization was progressively relaxed from G1 to G2. Double spots in G2 could reflect a similar gene organization of two chromatids. During mid-S phase, PCNA co-localized with some clustered genes (spots), indicating that rDNA replication occurs inside nucleoli and at different sites of the same locus simultaneously. The evaluation of the rDNA transcription units in 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB)-treated cells indicated a mean of 16 units per G1 nucleus and 25 units per G2 nucleus. For G1 and G2, the fine 3-D structure of nucleoli was reconstructed using ultrathin serial sections after specific contrast of DNA and RNA, digitization of the serial section images and computer-assisted 3-D architecture. Fibrillar centers (FCs) formed discrete structures (about 10 in G1 and 20 in G2) connected by a network of the dense fibrillar component. The 3-D arrangement of the FCs in G1 and G2 are similar to the rDNA spots. In conclusion, the architecture of the nucleoli during interphase reflects the distribution of the rDNA that is characterized by alternation of clustered and extended genes.

Animals↗

PubMed1999

Disruption of dynamic cell surface architecture of NIH3T3 fibroblasts by the N-terminal domains of moesin and ezrin: in vivo imaging with GFP fusion proteins.

Lamellipodia, filopodia, microspikes and retraction fibers are characteristic features of a dynamic and continuously changing cell surface architecture and moesin, ezrin and radixin are thought to function in these microextensions as reversible links between plasma membrane proteins and actin microfilaments. Full-length and truncated domains of the three proteins were fused to green fluorescent protein (GFP), expressed in NIH3T3 cells, and distribution and behaviour of cells were analysed by using digitally enhanced differential interference contrast (DIC) and fluorescence video microscopy. The amino-terminal (N-)domains of all three proteins localize to the plasma membrane and fluorescence recordings parallel the dynamic changes in cell surface morphology observed by DIC microscopy of cultured cells. Expression of this domain, however, significantly affects cell surface architecture by the formation of abnormally long and fragile filopodia that poorly attach and retract abnormally. Even more striking are abundant irregular, branched and motionless membraneous structures that accumulate during retraction of lamellipodia. These are devoid of actin, endogenous moesin, ezrin and radixin, but contain the GFP-labeled domain. While a large proportion of endogenous proteins can be extracted with non-ionic detergents as in untransfected control cells, >90% of N-moesin and >60% of N-ezrin and N-radixin remain insoluble. The minimal size of the domain of moesin required for membrane localization and change in behavior includes residues 1-320. Deletions of amino acid residues from either end result in diffuse intracellular distribution, but also in normal cell behavior. Expression of GFP-fusions of full-length moesin or its carboxy-terminal domain has no effect on cell behavior during the observation period of 6-8 hours. The data suggest that, in the absence of the carboxy-terminal domain, N-moesin, -ezrin and -radixin interact tightly with the plasma membrane and interfere with normal functions of endogeneous proteins mainly during retraction.

3T3 Cells↗

PubMed2005

Sarcomere length measurement permits high resolution normalization of muscle fiber length in architectural studies.

The use of sarcomere length to normalize fiber length in architectural studies is commonly practiced but has not been explicitly validated. Using mouse hindlimb muscles as a model system, ankle joints were intentionally set to angles ranging from 30 degrees to 150 degrees and their muscles fixed. Tibialis anterior (TA), extensor digitorum longus (EDL) and soleus muscles were removed and their raw fiber length measured. Sarcomere length was then measured for each fiber length sample and fiber length was normalized to a standard sarcomere length. As expected, raw fiber length was dependent on tibiotarsal angle (P < 0.0005 for all muscles, r2 range 0.22-0.61), while sarcomere length normalization eliminated the joint-angle dependent variation in fiber length (P > 0.24, r2 range 0.001-0.028). Similarly, one-way ANOVA revealed no significant differences in normalized fiber length among ankle angles for any of the three muscles (P > 0.1), regardless of animal size. To determine the resolution of the method, power calculations were performed. For all muscles studied, there was >90% chance of detecting a 15% fiber length difference among muscles and >60% chance of detecting fiber length differences as small as 10%. We thus conclude that the use of sarcomere length normalization in architectural studies permits resolution of fiber length variations of 15% and may even be effective at resolving 10% fiber length variations.

Analysis of Variance↗

PubMed2005

Muscle fiber angle, segment bulging and architectural gear ratio in segmented musculature.

The anatomical complexity of myomeres and myosepta has made it difficult to develop a comprehensive understanding of the relationship between muscle fiber architecture, connective tissue mechanics, and locomotor function of segmented axial musculature in fishes. The lateral hypaxial musculature (LHM) of salamanders is less anatomically complex and therefore a good system for exploring the basic mechanics of segmented musculature. Here, we derive a mathematical model of the LHM and test our model using sonomicrometry and electromyography during steady swimming in an aquatic salamander, Siren lacertina. The model predicts longitudinal segment strain well, with predicted and measured values differing by less than 5% strain over most of the range. Deviations between predicted and measured results are unbiased and probably result from the salamanders performing slight turns with associated body torsion in our unconstrained trackway swimming experiments. Model simulations of muscle fiber contraction and segment shortening indicate that longitudinal segment strain, for a given amount of muscle fiber strain, increases with increasing initial fiber angle. This increase in architectural gear ratio (AGR = longitudinal strain/fiber strain) is mediated by muscle fiber rotation; the higher the initial fiber angle, the more the fibers rotate during contraction and the higher the AGR. Muscle fiber rotation is additionally impacted by bulging in the dorsoventral (DV) and/or mediolateral (ML) dimensions during longitudinal segment shortening. In segments with obliquely oriented muscle fibers, DV bulging increases muscle fiber rotation, thereby increasing the AGR. Extending the model to include force and work indicates that force decreases with increasing initial muscle fiber angle and increasing DV bulging and that both longitudinal shortening and DV bulging must be included for accurate calculation of segment work.

Animals↗

PubMed1987

The effects of microenvironment on the redifferentiation of regenerating neurones: neurite architecture, acetylcholine receptors and Ca2+ channel distribution.

Severed adult neurones, which are capable of regrowth, encounter different microenvironments from those encountered during development. Moreover, adult neurones may respond in a different manner from developing neurones to the same environmental cues. Thus, the recovery of the integrative and transmission capabilities (which depend on the neuronal architecture, passive and active membrane properties, and synaptic receptor distribution) by a regenerating adult neurone may not be complete. In the present review, we examine several aspects of the outcome of the interaction between the microenvironment and regrowing neurones using the cockroach giant interneurones (GINs) as a model system. We demonstrate that whereas extrinsic cues govern the morphological redifferentiation and distribution of synaptic receptors, the distribution of voltage-dependent Ca2+ channels is to a large extent determined by intrinsic factors. The pathway of regrowth and the architecture of regenerating GINs were studied by examination of intracellularly stained fibres. The environments provided by the connectives and ganglia are different. The elongating sprouts in the connective appeared as smooth cylinders. Within the ganglionic domain, the main longitudinal sprouts emitted neurites which extended and branched into the neuropile. The local cues for branching of neurites were eliminated by freezing and thawing of the ganglia prior to the arrival of the growing tips. The failure to extend neurites under these conditions is attributed to the elimination of extrinsic signals for morphological redifferentiation of the fibres, since the same fibres emit neurites in anterior ganglia which have not been subjected to freezing and thawing. The distribution of acetylcholine receptors (AChRs) on the GINs was mapped by ionophoretic application of ACh. In both the intact and regenerating GINs receptors were located only on the neurites. Freezing and thawing of a ganglion eliminated the local signals for insertion and/or activation of AChRs on the neurites. Thus, both the morphological redifferentiation and the distribution of AChRs are affected by the microenvironment. Voltage-dependent Ca2+ channels were detected after intracellular injection of tetraethylammonium into the GIN and in the presence of tetrodotoxin (TTX) and Ba2+ in the extracellular space. The regrowing axon tips always revealed large barium action potentials independent of the CNS microenvironment. This observation is consistent with the hypothesis that Ca2+ plays an important role in the growth process. However, increased Ba2+ responsiveness was also observed in axonal segments proximal to the region of neuronal extension.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed2004

Effect of nicorandil, a K+ATP -channel opener, on coronary capillary architecture and volume after early myocardial ischemia-reperfusion--a 3-dimensional confocal laser microscopic study--.

BACKGROUND: Previous experimental and clinical studies have reported that nicorandil, an adenosine triphosphate (ATP)-sensitive K+ channel opener, exerts a beneficial effect on microvascular function and clinical outcome after myocardial ischemia. The present study assessed whether intravenous administration of nicorandil affects the 3-dimensional (3-D) architectural characteristics of capillaries and their volume after early myocardial reperfusion. METHODS AND RESULTS: Using the hearts of open-chest anesthetized rats, the left anterior descending artery was occluded for 7 min followed by reperfusion. Nicorandil or saline was infused intravenously during occlusion and reperfusion. The entire coronary microvasculature was filled with contrast medium after the hearts were arrested. Capillaries were observed 3-dimensionally by confocal laser scanning microscopy in both the control area and reperfused area of all samples. The capillary volume fraction was computed from the 3-D images. The reperfused area in both the nicorandil and saline groups showed characteristic architectural changes of the capillaries. Capillary volume fraction in the reperfused area was significantly reduced in saline group, compared with nicorandil group [12.7+/-7.2% vs 18.1 +/-5.3% (p<0.01)]. CONCLUSIONS: Intravenous nicorandil administration has a beneficial effect on capillary damage after reperfusion.

Adenosine Triphosphate↗

PubMed1975

Diet, mucosal architecture and epithelial cell production in the small intestine of specified-pathogen-free and conventional rats.

Upper jejunum and terminal ileum were examined in specified-pathogen-free (SPF), conventional and conventional after SPF rearing (ex-SPF) rats. The effect of 2 differential diets on the last 2 groups was examined. Ex-SPF rats had taller villi and deeper crypts than SPF rats, but similar crypt to villus ratios and cell production rates. Ex-SPF rats had similar crypt depth and jejunal villus height to conventional rats on the same diet, but taller ileal villi and a lower cell production rate. Even after 6-8 weeks, in a conventional environment, ex-SPF rat intestine was still not identical with conventional rat intestine. Diet had a significant effect on mucosal architecture, and a smaller effect on cell production rate. It is concluded that diet, microbiological status of colony of origin, and environment after weaning, can all affect mucosal architecture and epithelial cell production, and should be properly controlled in experimental studies.

Animal Feed↗

PubMed2001

Glucosylceramide synthesis inhibitors block pharmacologically induced dispersal of the Golgi and anterograde membrane flow from the endoplasmic reticulum: implication of sphingolipid metabolism in maintenance of the Golgi architecture and anterograde membrane flow.

PDMP (D,L-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol) and PPMP (D,L-threo-1-phenyl-2-hexadecanoylamino-3-morpholino-1-propanol), inhibitors of glucosylceramide synthesis, blocked brefeldin A (BFA)- and nordihydroguaiaretic acid-induced dispersal of the Golgi and trans Golgi network, and Golgi-derived vesicles were retained in the juxtanuclear region. PDMP and PPMP did not stabilize microtubules but blocked nocodazole-induced extensive fragmentation and dispersal of the Golgi, and large Golgi vesicles were retained in the juxtanuclear region. PPMP is a stronger inhibitor of glucosylceramide synthesis than PDMP, but PDMP showed a stronger activity against BFA-induced retrograde membrane flow. However, PPMP showed a stronger activity for Golgi disruption and inhibition of anterograde trafficking from the endoplasmic reticulum, and rebuilding of the Golgi architecture. Cumulatively, these results suggest that sphingolipid metabolism is implicated in maintenance of the Golgi architecture and anterograde membrane flow from the endoplasmic reticulum but not in Golgi dispersal induced by BFA.

Adenosine Triphosphate↗

PubMed1990

Effects of uvulopalatopharyngoplasty on sleep architecture and patterns of obstructed breathing.

In this retrospective study, 72 obstructive sleep apnea patients with polysomnograms taken before and after uvulopalatopharyngoplasty were evaluated. Postoperatively, there was a significant improvement of sleep architecture and respiratory indices. In addition, a second group of 17 patients also had position recordings with their polysomnograms. Time spent in supine and lateral sleep positions changed postoperatively. There was significant decrease of the apnea plus hypopnea index in the lateral position. This study indicates that there is significant improvement of sleep architecture and respiratory indices in the majority of patients after uvulopalatopharyngoplasty, particularly in the lateral sleep position.

Female↗

PubMed2001

Morphologic features suggestive of gluten sensitivity in architecturally normal duodenal biopsy specimens.

We studied small bowel biopsy specimens with architecturally normal villi from 78 adult patients with potential gluten sensitivity (GS) and correlated them with outcome to characterize morphologic features that would allow a pathologist to suggest GS. No patient had a previous GS diagnosis. Twelve study patients had GS. The mean number of intraepithelial lymphocytes (IELs) per 20 enterocytes from the tips of 5 random villi was significantly greater in GS than non-GS biopsy samples, but the groups overlapped significantly, making the number diagnostically useful only when markedly increased. Crypt mitoses counts had similar relationships. Twelve patients had an even distribution of IELs along villus sides and over tips (3/66 [5%] non-GS patients, 9/12 [75%] GS patients). Non-GS patients had a decrescendo pattern of IELs along the sides of villi. Architecturally normal small bowel biopsy specimens with an appreciable, continuous, even distribution of IELs along the sides and tips of villi and a mean of 12 or more IELs in the tips of several villi are suggestive of GS. Pathologists should be watchful for these morphologic features in small bowel biopsy specimens to suggest GS.

Adolescent↗

PubMed2000

Benign hematogone-rich lymphoid proliferations can be distinguished from B-lineage acute lymphoblastic leukemia by integration of morphology, immunophenotype, adhesion molecule expression, and architectural features.

Distinction of normal B-lymphoid proliferations including precursors known as hematogones from acute lymphoblastic leukemia (ALL) is critical for disease management. We present a multiparameter assessment of 27 bone marrow samples containing at least 25% hematogones (range, 25%-72%) by morphologic review. We used flow cytometry to evaluate B-cell differentiation antigen and adhesion molecule expression and immunohistochemistry on clot sections to evaluate architectural distribution. Flow cytometry revealed that intermediately differentiated cells (CD19+, CD10+) predominated, followed in frequency by CD20+, surface immunoglobulin-positive cells, with CD34+, terminal deoxynucleotidyl transferase (TdT)-positive cells as the smallest subset. Adhesion molecules (CD44, CD54) were expressed more heterogeneously compared with expression in acute lymphoblastic leukemia. Immunohistochemistry revealed that CD34+, TdT-positive cells were dispersed without significant clustering, while CD20+ cells exceeded CD34/TdT-positive cells in 24 of 25 cases. This multidisciplinary study demonstrates that hematogone-rich lymphoid proliferations exhibit a spectrum of B-lymphoid differentiation antigen expression with predominance of intermediate and mature B-lineage cells, heterogeneity of adhesion molecule expression, and nonclustered bone marrow architectural distribution.

Adolescent↗

PubMed2001

Intervertebral disc disorganization is related to trabecular bone architecture in the lumbar spine.

Cancellous bone morphometry was investigated in the sagittal plane of lumbar vertebrae using histoquantitation. The aim of this study was to identify variations in cancellous bone architecture at increasing states of intervertebral disc (IVD) disorganization after age adjustment and to investigate regional variations within the whole vertebral body. Measurements were taken of the ratio of bone volume (BV) to total volume (TV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), and trabecular number (Tb.N). Lumbar spines (T12-L5) of 19 men and 8 women were removed at autopsy from an adult sample with no clinical history of bone-related disease or histologically identifiable bone disease. It was found that degeneration of the IVD becomes more common with increasing age. After age-adjustment, significant increases in the proportion of BV/TV were observed in the presence of advancing IVD disorganization. Significant architectural changes were observed in the anterior regions of the vertebral body with increases in Tb.Th and Tb.N and decreases in Tb.Sp. Minimal alterations were found at posterior regions. Bone loss was observed in central regions (most distant from the cortex) as IVD disorganization increased through reduction in both Tb.N and Tb.Th. The BV/TV increase in anterior areas of the centrum may be a response to a redistribution of load to the vertebral body periphery as a result of IVD disorganization. It appears that trabecular morphology is related to the condition of the associated IVD, rather than being the sole consequence of a loss of BV/TV with age. This relationship could influence the occurrence of vertebral body crush fracture.

Adult↗

PubMed2002

Correction to the plant canopy gap-size analysis theory used by the Tracing Radiation and Architecture of Canopies instrument.

A plant canopy gap-size analyzer, the Tracing Radiation and Architecture of Canopies (TRAC), developed by Chen and Cihlar [Appl Opt. 34, 6211(1995)] and commercialized and by 3rd Wave Engineering (Nepean, Canada), has been used around the world to quantify the fraction of photosynthetically activeradiation absorbed by plant canopies, the leaf area index (LAI), and canopy architectural parameters. The TRAC is walked under a canopy along transects to measure sunflecks that are converted into a gap-size distribution. A numerical gap-removal technique is performed to remove gaps that are not theoretically possible in a random canopy. The resulting reduced gap-size distribution is used to quantify the heterogeneity of the canopy and to improve LAI measurements. It is explicitly shown here that the original derivation of the clumping index was missing a normalization factor. For a very clumped canopy with a large gap faction, the resulting LAI can be more than 100% smaller than previously estimated. A test case is used to demonstrate that the new clumping index derivation allows a more accurate change of LAI to be measured.

Journal Article↗

PubMed1990

Perceptual-components architecture for digital video.

A perceptual-components architecture for digital video partitions the image stream into signal components in a manner analogues to that used in the human visual system. These components consist of achromatic and opponent color channels, divided into static and motion channels, further divided into bands of particular spatial frequency and orientation. Bits are allocated to an individual band in accord with visual sensitivity to that band and in accord with the properties of visual masking. This architecture is argued to have desirable features such as efficiency, error tolerance, scalability, device independence, and extensibility.

Color Perception↗