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PubMed1992

MR imaging of the quadriceps tendon: normal layered configuration and its importance in cases of tendon rupture.

OBJECTIVE: The quadriceps tendon is a multilayered structure with separate layers arising from different muscle groups. We examined this layered configuration on MR images to determine if it is relevant in the evaluation of the traumatized quadriceps tendon. MATERIALS AND METHODS: Sagittal and axial T1-weighted MR images of 52 knees with normal tendons were reviewed to determine the number and the muscle origins of these layers. Five knees with traumatized quadriceps tendons were imaged with T1 and T2 weighting to determine the relationship between the layered configuration and the injury. RESULTS: All the normal quadriceps tendons had a laminated appearance with either four (6%), three (56%), or two (30%) layers. In 8%, the laminations were barely perceptible. The site of origin of each layer was consistent, and each favored a given muscle. The superficial layer originated from the posterior fascia of the rectus femoris muscle, and the deep layer originated from the anterior fascia of the midline vastus intermedius muscle. The middle layer(s) arose from the deep fascia separating the vastus medialis and the vastus lateralis muscles from the vastus intermedius muscle. When only two layers were present, the middle layer merged with the superficial and deep layers. In the injured tendon, complete rupture produced transection of all the layers. Incomplete ruptures were seen as focal discontinuities of individual layers; other layers remained intact. CONCLUSION: The existence of a layered configuration of the quadriceps tendon is significant in the MR examination of partial ruptures. It can help not only in detecting and localizing injuries but also in determining appropriate treatment based on the number or thickness of injured layers.

Female↗
PubMed2000

Effect of smear layers on the bonding of a self-etching primer to dentin.

PURPOSE: The objective of this work was to evaluate the effect of the absence and presence of smear layers on bonds made to dentin using a self-etching primer system, Clearfil SE Bond. MATERIALS AND METHODS: Dentin surfaces with different smear layer thickness were created from mid-coronal sound dentin in extracted, human third molars. The control group was cryofractured to create a bonding surface that was devoid of a smear layer. The experimental teeth were ground with wet 60-, 180- or 600-grit SiC paper. They were bonded using SE Bond, followed by resin composite buildups. After 1 day, bonded specimens were sectioned into multiple 1- x 1-mm beams. Microtensile bond strengths were determined and the results analyzed with ANOVA and the Student Neuman Keuls test. Fractographic study of cross sections of failed interfaces from the dentin side of representative beams was performed using both SEM and TEM. RESULTS: SE Bond produced high bond strengths (ca 50 MPa) to both smear layer-free and smear layer-covered dentin. SEM examination was inadequate to define the exact nature of interfacial failures. TEM observations demonstrated a thin (ca 400 to 500 nm) hybrid layer in the fractured dentin and thicker (ca 1 to 4 microns) hybrid layers on smear layer-covered dentin. This included a thick, hybridized smear layer and a thin, underlying true hybrid layer in the intact dentin. Separation of the two hybrid layers was not evident in interfacial failures. CONCLUSION: Self-etching primers create thin hybrid layers that incorporate the smear layer. This study shows that formation of true hybrid layers occurs irrespective of smear layer thickness and that both hybrid layers may function as a unit during loading without separation. identification of secondary cracks from TEM fractographic analysis exemplifies the complex reaction to tensile stresses in multilayered joint systems that comprise materials of variable compliance.

Acid Etching, Dental↗
PubMed1983

The formation and growth of the cortical layers in the cerebellum of the opossum.

The development of the cerebellar cortex in the opossum was analyzed in Nissl-stained sections using qualitative and quantitative methods. The young of the opossum are born 12-13 days after conception and mature for approximately 85 days in an external pouch providing an excellent model for embryological studies. Qualitative observations of cerebellar growth were made from birth to postnatal day (PN) 19. At birth the opossum cerebellar anlage can be divided into two layers, a ventricular layer and an intermediate layer; histologically his is comparable to the rat cerebellar anlage at embryonic day 13 (Altman and Bayer 1978) and the human cerebellar anlage prior to the seventh embryonic week (Rakic and Sidman 1970). By PN 3 the cerebellar anlage consists of five layers: the ventricular layer, the ventral intermediate layer, the acellular layer, the dorsal intermediate layer and the marginal layer. The external granular layer begins migrating over the dorsal surface of the cerebellum at PN 5. The immature Purkinje cell layer is first seen at PN 12 and is subsequently arranged as four clusters between PN 12 and PN 22. At PN 19 the opossum cerebellum is comparable to the rat cerebellum at birth (Korneliussen 1968c). A quantitative analysis of cerebellar growth was performed between PN 17 and PN 77 using vermal sections. The area and thickness of each of the cortical layers was determined from five vermal sagittal sections using two methods; a Zeiss Videoplan and a point counting system. The external granular layer increases in area from PN 17 to PN 75, however its maximal width is achieved between PN 19 and PN 33. The persistence of the EGL until after PN 105 suggests that synaptic contacts between granule cell axons and Purkinje cells may continue to form after PN 77 when the Purkinje cell is mature based on Golgi and fine structural features (Laxson and King 1983). Between PN 17 and PN 77 the area of the molecular layer and the area of the internal granular layer increase at a more rapid rate than the other cerebellar layers. The maturation of the cerebellum in the opossum is a lengthy process lasting approximately 77 days in comparison to rodent cerebellar growth which requires about 25 days (Korneliussen 1968c). Also, the entire process of cortical lamination occurs after birth while the opossum is maturing in an external pouch.

Animals↗
PubMed2004

Input from the presubiculum to dendrites of layer-V neurons of the medial entorhinal cortex of the rat.

The entorhinal cortex (EC) and the hippocampus are reciprocally connected. Neurons in the superficial layers of EC project to the hippocampus, whereas deep entorhinal layers receive return connections. In the deep layers of EC, pyramidal neurons in layer V possess apical dendrites that ascend towards the cortical surface through layers IIII and II. These dendrites ramify in layer I. By way of their apical dendrites, such layer-V pyramidal cells may be exposed to input destined for the superficial entorhinal neurons. A specific and dense fiber projection that typically ends in superficial entorhinal layers of the medial EC originates in the presubiculum. To investigate whether apical dendrites of deep entorhinal pyramidal neurons indeed receive input from this projection, we injected the anterograde tracer PHA-L in the presubiculum or we lesioned the presubiculum, and we applied in the same experiments the tracer Neurobiotin trade mark pericellularly in layer V of the medial EC of 17 rats. PHA-L labeled presubiculum axons in the superficial layers apposing apical segments of Neurobiotin labeled layer-V cell dendrites were studied with a confocal fluorescence laserscanning microscope. Axons and dendrites were 3D reconstructed from series of confocal images. In cases in which the presubiculum had been lesioned, material was investigated in the electron microscope. At the confocal fluorescence microscope level we found numerous close contacts, i.e. appositions of boutons on labeled presubiculum fibers with identified dendrites of layer-V neurons. In the electron microscope we observed synapses between degenerating axon terminals and spines on dendrites belonging to layer-V neurons. Hence we conclude that layer-V neurons receive synaptic contacts from presubiculum neurons. These findings indicate that entorhinal layer-V neurons have access to information destined for the superficial layers and eventually the hippocampal formation. At the same time, they have access to the hippocampally processed version of that information.

Animals↗
PubMed1997

Membrane-associated molecules regulate the formation of layer-specific cortical circuits.

The columnar organization of the mammalian neocortex is based on radially oriented axon collaterals which precisely link cells from distinct cortical layers. During development, these interlaminar connections are specific from their initial outgrowth: collaterals form only in the target layers and there are no transient axonal collaterals in the nontarget layers. To examine whether positional cues within individual cortical layers regulate the laminar specificity of collateral formation, explants of cells destined for different cortical layers were cultured on membranes prepared from target and nontarget layers. Axonal growth and branching were examined on homogeneous membrane substrates and on alternating stripes of membranes from different layers. Results show that axons branch preferentially on membrane substrates from those layers that they would target in vivo. In addition, when cortical axons were given a choice to grow on membranes from either their target or their nontarget layer, they exhibited a clear preference for the target layers. This indicates that membrane-associated cues confined to individual layers regulate the formation of collaterals of cortical axons and restrict their growth to their target layers. Heat inactivation of membranes from target layers resulted in reduced axonal branching. The same manipulation of membranes from nontarget layers increased axonal branching for one population of cortical neurons. Taken together, these results suggest that membrane-associated molecules confined to individual layers induce and prevent the formation of axon collaterals in distinct populations of cortical neurons. Thus, the expression of layer-specific cues provides important constraints for the remodeling of local circuits during cortical development.

Animals↗
PubMed2000

The connections of layer 4 subdivisions in the primary visual cortex (V1) of the owl monkey.

The primary visual cortex (V1) of primates receives signals from parallel lateral geniculate nucleus (LGN) channels. These signals are utilized by the laminar and compartmental [i.e. cytochrome oxidase (CO) blob and interblob] circuitry of V1 to synthesize new output pathways appropriate for the next steps of analysis. Within this framework, this study had two objectives: (i) to analyze the con- nections between primary input and output layers and compartments of V1; and (ii) to determine differences in connection patterns that might be related to species differences in physiological properties in an effort to link specific pathways to visual functions. In this study we examined the intrinsic interlaminar connections of V1 in the owl monkey, a nocturnal New World monkey, with a special emphasis on the projections from layer 4 to layer 3. Interlaminar connections were labeled via small iontophoretic or pressure injections of tracers [horseradish peroxidase, biocytin, biotinylated dextrine amine (BDA) or cholera toxin subunit B conjugated to colloidal gold particles]. Our most significant finding was that layer 4 (4C of Brodmann) can be divided into three tiers based upon projections to the superficial layers. Specifically, we find that 4alpha (4Calpha), 4beta (4Cbeta) and 4ctr send primary projections to layers 3C (4B), 3Bbeta (4A) and 3Balpha (3B), respectively. Examination of laminar structure with Nissl staining supports a tripartite organization of layer 4. The cortical output layer above layer 3Balpha (3B) (e.g. layer 3A) does not appear to receive any direct connections from layer 4 but receives heavy input from layers 3Balpha (3B) and 3C (4B). Some connectional differences also were observed between the subdivisions of layer 3 and the infragranular layers. No consistent differences in connections were observed that distinguished CO blobs from interblobs or that could be correlated with differences in visual lifestyle (nocturnal versus diurnal) when compared with connectional data in other primates. Re-examination of data from previous studies in squirrel and macaque monkeys suggests that the tripartite organization of layer 4 and the unique projection pattern of layer 4ctr are not restricted to owl monkeys, but are common to a number of primate species.

Animals↗
PubMed1990

Antigenic differences among Campylobacter fetus S-layer proteins.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of S-layer proteins extracted from Campylobacter fetus strains by using acid glycine buffer showed that the predominant S-layer proteins of different strains had subunit molecular weights in the range of 90,000 to 140,000. Electron microscopy revealed oblique S-layer lattices with a spacing of approximately 5.6 nm (gamma = 75 degrees) on wild-type strains VC1, VC119, VC202, and VC203. Three variants of C. fetus VC119 producing a predominant S-layer subunit protein of different molecular weight (Mr) from that of the parent were also examined. Each variant produced an oblique lattice morphologically indistinguishable from that of the parent. Amino-terminal sequence analysis showed that the S-layer proteins of the VC119 parent and variants were identical up to residue 18 and that this sequence differed from but was related to the first 16 N-terminal residues shared by the S-layer proteins of the three other wild-type C. fetus isolates. Western immunoblot analysis with an antiserum prepared to the VC119 protein and an antiserum prepared to C. fetus 84-40 LP (Z. Pei, R. T. Ellison, R. V. Lewis, and M. J. Blaser, J. Biol. Chem. 263:6416-6420, 1988) showed that strains of C. fetus were capable of producing S-layer proteins with at least four different antigenic specificities. Immunoelectron microscopy with antiserum to the VC119 S-layer protein showed that C. fetus cultures contained cells with immunoreactive oblique S-layer lattices as well as cells with oblique S-layer lattices which did not bind antibody. This suggests that C. fetus S-layer proteins undergo antigenic variation. Thermal denaturation experiments indicated that the antigenicity conferred by the surface-exposed C. fetus S-layer epitopes was unusually resistant to heat, and the thermal stability appeared to be due to the highly organized lattice structure of the S. layer. Protease digestion of purified VC119 S-layer protein revealed a trypsin-, chymotrypsin-, and endoproteinase Glu-C-resistant domain with an apparent Mr of 110,000, which carried the majority of the epitopes of the S-layer protein, and a small enzyme-sensitive domain. The trypsin- and chymotrypsin-resistant polypeptides shared an overlapping sequence which differed from the N-terminal sequence of the intact S-layer protein.

Amino Acid Sequence↗
PubMed1996

Evidence that an N-terminal S-layer protein fragment triggers the release of a cell-associated high-molecular-weight amylase in Bacillus stearothermophilus ATCC 12980.

During growth on starch medium, the S-layer-carrying Bacillus stearothermophilus ATCC 12980 and an S-layer-deficient variant each secreted three amylases, with identical molecular weights of 58,000, 122,000, and 184,000, into the culture fluid. Only the high-molecular-weight amylase (hmwA) was also identified as cell associated. Extraction and reassociation experiments showed that the hmwA had a high-level affinity to the peptidoglycan-containing layer and to the S-layer surface, but the interactions with the peptidoglycan-containing layer were stronger than those with the S-layer surface. For the S-layer-deficient variant, no changes in the amount of cell-associated and free hmwA could be observed during growth on starch medium, while for the S-layer-carrying strain, cell association of the hmwA strongly depended on the growth phase of the cells. The maximum amount of cell-associated hmwA was observed 3 h after inoculation, which corresponded to early exponential growth. The steady decrease in cell-associated hmwA during continued growth correlated with the appearance and the increasing intensity of a protein with an apparent molecular weight of 60,000 on sodium dodecyl sulfate gels. This protein had a high-level affinity to the peptidoglycan-containing layer and was identified as an N-terminal S-layer protein fragment which did not result from proteolytic cleavage of the whole S-layer protein but seems to be a truncated copy of the S-layer protein which is coexpressed with the hmwA under certain culture conditions. During growth on starch medium, the N-terminal S-layer protein fragment was integrated into the S-layer lattice, which led to the loss of its regular structure over a wide range and to the loss of amylase binding sites. Results obtained in the present study provide evidence that the N-terminal part of the S-layer protein is responsible for the anchoring of the subunits to the peptidoglycan-containing layer, while the surface-located C-terminal half could function as a binding site for the hmwA.

Amylases↗
PubMed1998

Contribution of supragranular layers to sensory processing and plasticity in adult rat barrel cortex.

Contribution of supragranular layers to sensory processing and plasticity in adult rat barrel cortex. J. Neurophysiol. 80: 3261-3271, 1998. In mature rat primary somatic sensory cortical area (SI) barrel field cortex, the thalamic-recipient granular layer IV neurons project especially densely to layers I, II, III, and IV. A prior study showed that cells in the supragranular layers are the fastest to change their response properties to novel changes in sensory inputs. Here we examine the effect of removing supragranular circuitry on the responsiveness and synaptic plasticity of cells in the remaining layers. To remove the layer II + III (supragranular) neurons from the circuitry of barrel field cortex, N-methyl--aspartate (NMDA) was applied to the exposed dura over the barrel cortex, which destroys those neurons by excitotoxicity without detectable damage to blood vessels or axons of passage. Fifteen days after NMDA treatment, the first responsive cells encountered were 400-430 micrometers below the pial surface. In separate cases triphenyltetrazolium chloride (TTC), a vital dye taken up by living cells, was absent from the lesion area. Cytochrome oxidase (CO) activity was absent in the first few tangential sections through the barrel field in all cases before arriving at the CO-dense barrel domains. These findings indicate that the lesions were quite consistent from animal to animal. Controls consisted of applying vehicle without NMDA under similar conditions. Responses of D2 barrel cells were assessed for spontaneous activity and level of response to stimulation of the principal D2 whisker and four surround whiskers D1, D3, C2, and E2. In two additional groups of animals treated in the same way, sensory plasticity was assessed by trimming all whiskers except D2 and either D1 or D3 (called Dpaired) for 7 days before recording cortical responses. Such whisker pairing normally potentiates D2 barrel cell responses to stimulation of the two intact whiskers (D2 + Dpaired). After NMDA lesions, cortical cells still responded to all whiskers tested. Cells in lesioned cortex showed reduced response amplitude compared with sham-operated controls to all D-row whiskers. In-arc surround whisker (C2 or E2) responses were normal. Spontaneous activity did not change significantly in any remaining layer at the time tested. Modal latencies to stimulation of principal D2 or surround D1 or D3 whiskers showed no significant change after lesioning. These findings indicate that there is a reasonable preservation of the response properties of layer IV, V, VI neurons after removal of layer II-III neurons in this way. Whisker pairing plasticity in layer IV-VI D2 barrel column neurons occurred in both lesioned and sham animals but was reduced significantly in lesioned animals compared with controls. The response bias generated by whisker trimming (Dpaired/Dcut + Dpaired ratio) was less pronounced in NMDA-lesioned than sham-lesioned animals. Proportionately fewer neurons in layer IV (52 vs. 64%) and in the infragranular layers (55 vs. 68%) exhibited a clear response bias to paired whiskers. We conclude that receptive-field plasticity can occur in layers IV-VI of barrel cortex in the absence of the supragranular layer circuitry. However, layer I-III circuitry does play a role in normal receptive-field generation and is required for the full expression of whisker pairing plasticity in granular and infragranular layer cells.

Animals↗
PubMed2003

[Expression of mRNAs for GHR, IGF-IR, FSHR and LHR in granulosa and theca layers of ovarian follicles of Shaoxing ducks].

Expression of genes encoding growth hormone receptor (GHR), type I insulin-like growth factor receptor (IGF-IR), follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) was measured in granulosa and theca layers of the largest (F1), third largest (F3), fifth largest (F5) preovulatory follicles and large white follicles (LWF) in the ovary of Shaoxing ducks, with relative quantitative reverse transcription-polymerase chain reaction (RT-PCR) using beta-actin as an internal standard. The results showed that GHR mRNA was more abundant in theca layer than in granulosa layer in all the follicles investigated, in theca layer, the LWF follicle expressed highest the level of GHR mRNA, while no differences in granulosa layer were observed among follicles at different stages of development. In contrast, the expression of IGF-IR mRNA in theca layer was evidently lower than that in granulosa layer, but no significant changes were found among different stages of follicles in either layers, except that LWF trended to express higher IGF-IR mRNA in the theca layer compared to other preovulatory follicles. Similar to IGF-IR mRNA, FSHR mRNA was more abundant in granulosa layer and no significant differences were found among different stages of follicles in either layers. In contrast, the expression of LHR mRNA followed a different pattern. In the theca layer, the expression level maintained consistent, while in the granulosa layer, a significant stepwise increase was observed as the follicles matured, resulting in higher mRNA level of LHR in the granulosa layer of largest follicles (F1), but lower mRNA level of LHR in the granulosa layer of smaller follicles (F5 and LWF). These results suggest that GH and IGF-I regulate ovary function by respective dominant sites in the follicles, and they cooperate with LH and FSH to regulate the follicle development via their respective receptors. The high expression of LHR mRNA in granulosa layer of large follicles might be related to the establishment of the follicle hierarchy and ovulation.

Actins↗
PubMed2001

Aggressiveness of contemporary self-etching systems. I: Depth of penetration beyond dentin smear layers.

OBJECTIVE: This study examined, with the use of transmission electron microscopy (TEM), the aggressiveness of three self-etching adhesive systems in penetrating dentin smear layers of different thickness. METHODS: Dentin disks were produced from extracted human third molars. For the control group, the middle dentin surface was cryofractured to create a bonding surface that was devoid of a smear layer. The experimental teeth were polished with wet 600 or 60-grit SiC paper to produce bonding surfaces with thin and thick smear layers. They were bonded using one of the three self-etching systems: Clearfil Mega Bond (Kuraray), Non-Rinse Conditioner and Prime&Bond NT (Dentsply DeTrey) and Prompt L-Pop (ESPE). Bonded specimens were then demineralized and embedded in epoxy resin for TEM examination. RESULTS: For Mega Bond, thin authentic hybrid layers between 0.4-0.5 microm were found. Smear layer and smear plugs were retained as part of the hybridized complex. For Non-Rinse Conditioner/Prime&Bond NT, the authentic hybrid layers were between 1.2-2.2 microm thick. Smear layer and smear plugs were completely dissolved in dentin with thin smear layers, but were partially retained as part of the hybridized complex in those with thick smear layers. For Prompt L-Pop, authentic hybrid layers were 2.5-5 microm thick and smear layer and smear plugs were completely dissolved even in dentin with thick smear layers. SIGNIFICANCE: Contemporary self-etching systems may be classified as mild, moderate and aggressive based on their ability to penetrate dentin smear layers and their depth of demineralization into the subsurface dentin. The more aggressive system completely solubilized the smear layer and smear plugs and formed hybrid layers with a thickness approaching those of phosphoric acid conditioned dentin.

Acid Etching, Dental↗
PubMed1998

Reattachment rate of human retinal pigment epithelium to layers of human Bruch's membrane.

OBJECTIVES: To determine the reattachment rate of human retinal pigment epithelium (RPE) to different layers of human Bruch's membrane (BM). METHODS: Explants of BM were prepared from 10 human cadaver eyes by removing native RPE. The RPE basal lamina, inner collagenous layer, elastin layer, and outer collagenous layer were exposed by sequentially removing each apical layer by mechanical or enzymatic means. First-passage human RPE was plated onto the surface and the RPE reattachment rate to each layer of BM was determined. RESULTS: Retinal pigment epithelial cell reattachment was highest to the inner aspects of BM and decreased as deeper layers of BM were exposed (ie, reattachment rate to basal lamina was higher than to the inner collagenous layer, which was higher than to the elastin layer, which was higher than to the outer collagenous layer). The reattachment rate to the inner collagenous layer, elastin layer, and outer collagenous layer harvested from elderly donors (age >60 years) was less than the reattachment rate to corresponding layers harvested from younger (age <50 years) donors. CONCLUSIONS: Retinal pigment epithelial cell reattachment depends on the anatomical layer of BM present in the host tissue. Age-related changes in BM may interfere with RPE reattachment. Our observations may have implications for understanding the pathogenesis of age-related macular degeneration and its potential treatment with RPE transplantation techniques.

Adult↗
PubMed2003

Stress distribution in the layered wall of the rat oesophagus.

The topic of this study is to obtain the constitutive equations for two layers in the oesophagus from inflation experiment of each layer in the in vivo state and to calculate the corresponding stress distribution referenced to a new stress-free state for multi-layered tissue. The oesophagus is treated as a two-layered structure consisting of an inner submucosa layer and an outer muscle layer. An anisotropic exponential pseudo-strain energy density function is used as the constitutive equation to fit the experimental loading curve and for the calculation of the stress distribution in each layer. Significant differences in the constitutive parameters and zero-stress parameter were found between the submucosa layer and the muscle layer. The stress in the submucosa layer was higher than that in the muscle layer and intact (non-separated) wall under the same loading conditions. The submucosa layer was stiffer than the muscle layer and the intact wall. The zero-stress state of the intact oesophagus and the zero-stress state after separation were used as a reference to compare the stress-strain distributions. The residual strain was discontinuous when using the zero-stress state after separation as a reference. Furthermore, the circumferential stress when using the zero-stress state of the intact wall as a reference was about 100 times higher than that referenced to the zero-stress state after separation. Hence, it is important to use the zero-stress state obtained after the layer separation as the stress-free state in the study of multi-layered tissue.

Animals↗
PubMedDate not supplied

The functional organization of area V2, I: specialization across stripes and layers.

We used qualitative tests to assess the sensitivity of 1043 V2 neurons (predominantly multiunits) in anesthetised macaque monkeys to direction, length, orientation, and color of moving bar stimuli. Spectral sensitivity was additionally tested by noting ON or OFF responses to flashed stimuli of varied size and color. The location of 649 units was identified with respect to cycles of cytochrome oxidase stripes (thick-inter-thin-inter) and cortical layer. We used an initial 8-way stripe classification (4 stripes, and 4 "marginal" zones at interstripes boundaries), and a 9-way layer classification (5 standard layers (2-6), and 4 "marginal" strata at layer boundaries). These classes were collapsed differently for particular analyses of functional distribution; the main stripe-by-layer analysis was performed on 18 compartments (3 stripes x 6 layers). We found direction sensitivity only within thick stripes, orientation sensitivity mainly in thick stripes and interstripes, and spectral sensitivity mainly in thin stripes. Positive length summation was relatively more frequent in thick stripes and interstripes, and negative length/size summation in thin stripes. All these "majority" characteristics of stripes were most prominent in layers 3A and 3B. By contrast, "minority" characteristics (e.g. spectral sensitivity in thick stripes; positive size summation in thin stripes) tended to be most frequent in the outer layers, that is, layers 2 and 6. In consequence, going by the four functions tested, the distinctions between stripes were maximal in layer 3, moderate in layer 2, and minimal in layer 6. Pooling all layers, there was some indication of asymmetry in the stripe cycle, in that thin stripe characteristics (spectral sensitivity, orientation insensitivity, and negative size summation) were also evident in the marginal zone and interstripe immediately lateral to a thin stripe, but less so medially. Within thin stripes, spectral and orientation selectivities were negatively correlated; this was still more accentuated amongst the minority spectrally tuned cells of thick stripes, but absent from interstripes, where these two properties were randomly assorted. Directional and spectral sensitivities were each coupled to negative size summation, but not to each other. We conclude that these functional characteristics of stripes are consistent with segregated, specialized pathways ascending through their middle layers, whilst the outer layers, 1, 2, and 6, utilize feedback from higher areas to adopt a more integrative role.

Animals↗
PubMedDate not supplied

Thalamic control of cat area-18 supragranular layers: simple cells, complex cells, and cells projecting to the lateral suprasylvian visual area.

The goal of this study was to determine the effects of inactivating layer A or the C layers of the cat lateral geniculate nucleus on the supragranular layers of area 18, including cells antidromically activated from the lateral suprasylvian visual area (LS). Isolated cells were visually driven via the contralateral eye while the retinotopically corresponding regions of layer A or, in some cases, the C layers were reversibly inactivated with injections of cobaltous chloride. Simple cells were frequently encountered and were on average more dependent upon layer A than were complex cells, a result qualitatively similar to that found previously in area 17 (Malpeli, 1983; Malpeli et al., 1986). However, the influence of the C layers on area 18 was much more apparent than for area 17. In area 18, as in area 17, the dependence of simple cells on particular geniculate layers appears to follow the terminal patterns of the major direct geniculate inputs. Those simple cells most dependent on layer A were located in lower layer 3. Simple cells in upper layer 3, like complex cells, showed little dependence on layer A, but were strongly dependent upon the C layers. All cells antidromically activated from LS were simple cells with rapidly conducting axons. They had, on average, the same moderately strong dependence on layer A as the patches of LS receiving area 18 input (Lee et al., 1997), supporting the conclusion that the influence of layer A in these patches is largely transmitted via association inputs from area 18. These results demonstrate that simple cells play a major role in association pathways.

Animals↗
PubMed2001

Dual coupling of opioid receptor-like (ORL1) receptors to adenylyl cyclase in the different layers of the rat main olfactory bulb.

The coupling of opioid receptor-like (ORL1) receptors to adenylyl cyclase has been investigated in specific layers of the rat main olfactory bulb. Membranes prepared from the olfactory nerve-glomerular layer (ON-G layer), external plexiform layer (EP layer) and granule cell layer (GR layer) displayed specific binding sites for [(3)H]-nociceptin/orphanin FQ ([(3)H]Noc/OFQ). In each layer, the presence of high-and low-affinity binding sites, with K(D) values in the picomolar and nanomolar range, respectively, was detected. The binding of [(3)H]Noc/OFQ was displaced by unlabelled Noc/OFQ, but not by opioid antagonists. In each layer, Noc/OFQ significantly stimulated [(35)S]GTPgammaS binding with nanomolar potencies. In ON-G layer, Noc/OFQ inhibited basal adenylyl cyclase activity and the enzyme stimulations by corticotropin releasing hormone (CRH), Ca(2+)/calmodulin (Ca(2+)/CaM) and forskolin (FSK). In EP layer, Noc/OFQ inhibited Ca(2+)/CaM-and FSK-stimulated enzyme activities. Conversely, in GR layer the peptide stimulated basal cyclase activity and potentiated the enzyme activation by CRH. The Noc/OFQ stimulation was counteracted by the GDP-bound form of the alpha subunit of transducin and was mimicked by transducin betagamma subunits. In the same tissue layer, Ca(2+)/CaM-and FSK-stimulated enzyme activities were inhibited. Naloxone failed to antagonize all the actions of Noc/OFQ. Western blot and RT-PCR analysis revealed the expression of Ca(2+)-insensitive and -sensitive adenylyl cyclases in the three layers. These results demonstrate that in rat main olfactory bulb ORL1 receptors can differentially affect distinct forms of adenylyl cyclase in a layer specific manner.

Adenylyl Cyclases↗
PubMed1970

Quantitation, chemical characteristics, and ultrastructure of the three outer cell wall layers of a gram-negative bacterium.

The cell wall of the gram-negative marine pseudomonad (American Type Culture Collection 19855) consists of three layers: the loosely bound outer layer, the outer double-track layer, and the underlying layer. These three layers constitute 4.7, 7.9, and 6.1%, respectively, of the dry weight of the whole cells. All three layers contained protein, lipid, and carbohydrate. The loosely bound outer layer and underlying layer were lower in protein and lipid and higher in amino and nonamino carbohydrate than the outer double-track layer. All three layers contained proteins with similar amino acid compositions. Minicell-like forms attached to the ends of cells were separated with and fractionated from the units of loosely bound outer layer. Examination of negatively stained preparations by electron microscopy revealed the loosely bound outer layer to be composed largely of units ranging from 400 to 1000 nm in diameter. The outer double-track layer, by the same technique, appeared as large, usually rounded sheets, each with a distinct rim. Washing this layer changed the gross chemical composition but did not affect the bimolecular leaflet appearance in thin sections. The underlying layer, when negatively stained, appeared to be composed of a heterogeneous mixture of particles differing in size and shape. It was separated by gel filtration into a large fraction with a molecular weight range in excess of 20 x 10(6) to 40 x 10(6) and a small fraction with a lower range of molecular weight. The larger fraction contained both protein and hexosamine, whereas the smaller one contained protein and only traces of hexosamine. A cytochrome-like pigment separated with this latter fraction.

Journal Article↗
PubMed1995

Physical and functional S-layer reconstitution in Aeromonas salmonicida.

The various functions attributed to the S-layer of Aeromonas salmonicida have been previously identified by their conspicuous absence in S-layer-defective mutants. As a different approach to establish the multifunctional nature of this S-layer, we established methods for reconstitution of the S-layer of A. salmonicida. Then we investigated the functional competence of the reconstituted S-layer. S-layers were reconstituted in different systems: on inert membranes or immobilized lipopolysaccharide (LPS) from purified S-layer protein (A-protein) or on viable cells from either A-protein or preassembled S-layer sheets. In the absence of divalent cations and LPS, purified A-protein in solution spontaneously assembled into tetrameric oligomers and, upon concentration by ultrafiltration, into macroscopic, semicrystalline sheets formed by oligomers loosely organized in a tetragonal arrangement. In the presence of Ca2+, purified A-protein assembled into normal tetragonal arrays of interlocked subunits. A-protein bound with high affinity (Kd, 1.55 x 10(-7) M) and specificity to high-molecular-weight LPS from A. salmonicida but not to the LPSs of several other bacterial species. In vivo, A-protein could be reconstituted only on A. salmonicida cells which contained LPS, and Ca2+ affected both a regular tetragonal organization of the reattached A-protein and an enhanced reattachment of the A-protein to the cell surface. The reconstitution of preformed S-layer sheets (produced by an S-layer-secreting mutant) to an S-layer-negative mutant occurred consistently and efficiently when the two mutant strains were cocultured on calcium-replete solid media. Reattached A-protein (exposed on the surface of S-layer-negative mutants) was able to bind porphyrins and an S-layer-specific phage but largely lacked regular organization, as judged by its inability to bind immunoglobulins. Reattached S-layer sheets were regularly organized and imparted the properties of porphyrin binding, hydrophobicity, autoaggregation, adherence to and invasion of fish macrophages and epithelial cells, and resistance to macrophage cytotoxicity. However, cells with reconstituted S-layers were still sensitive to complement and insensitive to the antibiotics streptonigrin and chloramphenicol, indicating incomplete functional reconstitution.

Aeromonas↗