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PubMed1998

Direct projections from the entorhinal cortical layers to the dentate gyrus, hippocampus, and subicular complex in the cat.

Projections from each layer of the entorhinal cortex (EC) of the cat were traced to the dentate gyrus (DG), Ammon's horn (CA), prosubiculum (ProSb), subiculum (Sb), presubiculum (PreSb) and parasubiculum (ParaSb); the anterograde or retrograde labeling method was used after stereotaxic injection of wheat germ agglutinin-horseradish peroxidase, cholera toxin B subunit, or Phaseolus vulgaris leucoagglutinin. On the side ipsilateral to the tracer-injection, layer II of the EC projected most abundantly to the outer half of the molecular layer (ML) of the DG, less abundantly to the almost entire thickness of the stratum lacunosum-moleculare (SLM) of CA2-3, moderately to the almost entire thickness of the SLM of CA1, and less to the outer part of the ML of the ProSb and Sb. Layer III projected abundantly to the almost entire thickness of the SLM of CA1 and outer part of the ML of the ProSb and Sb, and sparsely to the SLM of CA2-3. Layer IV projected sparsely to the pyramidal cell layer of the ProSb and Sb; Layer IV of the medial part (toward the ParaSb) of the EC projected further to the ML of the DG. Layer VI projected sparsely to the outer part of the ML of the DG, almost entire thickness of the SLM of CA1-3, and outer part of the ML of the ProSb and Sb. More temporal parts of the hippocampal region received the projections from progressively more medial and more rostral parts of layers II and III, and from progressively more rostral parts of layers IV and VI. The ML of the PreSb and ParaSb received projections from all layers of the medial part of the ipsilateral EC. The SLM of CA1 and ML of the ProSb, Sb and ParaSb received projections from layer II and/or III of the contralateral medial entorhinal area.

Animals↗

PubMed2003

Biomass accumulation and carbon utilization in layered sand filter biofilm systems receiving milk fat and detergent mixtures.

This research evaluated utilization of organic compounds and biomass accumulation in three different layered sand filter biofilm systems renovating a mixture of detergent and milk fat. Organic compounds were measured as chemical oxygen demand (COD) and biomass as ATP. A two-layer sand filter with coarse sand on the top and fine sand at the bottom; a three-layer sand filter with pea gravel on the top, coarse sand in the middle, and fine sand at the bottom; and a two-filters in-series sand filter biofilm system each received mixtures of butterfat and detergent. While the three-layer sand filter system had greater COD utilization capacity due to a longer filter run, the two-filters in-series sand filter system had greater COD utilization efficiency. The COD utilization was highly associated with the period of filter run to clogging. The decrease of COD utilization rate could lead to the clogging of sand filters. To maintain long filter runs without clogging, a COD loading rate of 2100 mg O(2)/m(2)/h was recommended for three-layer sand filters and 1800 mg O(2)/m(2)/h for two-layer sand filters for high fat content wastewater. In the two-layer sand filters and the three-layer sand filters, biomass did not seem to accumulate in any particular layer on a unit volume of sand. However, based on unit surface area, the coarse sand layer accumulated more biomass than the fine sand layer and the pea gravel layer.

Adenosine Triphosphate↗

PubMed2006

CdTe electrodeposition on InP(100) via electrochemical atomic layer epitaxy (EC-ALE): studies using UHV-EC.

The II-VI compound semiconductor CdTe was electrodeposited on InP(100) surfaces using electrochemical atomic layer epitaxy (EC-ALE). CdTe was deposited on a Te-modified InP(100) surface using this atomic layer by atomic layer methodology. The deposit started with formation of an atomic layer of Te on the InP(100) surface, as Cd was observed not to form an underpotential deposition (UPD) layer on InP(100), although it was found to UPD on Te atomic layers. On the In-terminated 'clean' InP(100) surface, Te was deposited at -0.80 V from a 0.1 mM solution of TeO2, resulting in formation of a Te atomic layer and some small amount of bulk Te. The excess bulk Te was then removed by reduction in blank solution at -0.90 V, leaving a Te atomic layer. Given the presences of the Te atomic layer, it was then possible to form an atomic layer of Cd by UPD at -0.58 V to complete the formation of a CdTe monolayer by EC-ALE. That cycle was then repeated to demonstrate the applicability of the cycle to the formation of CdTe nanofilms. Auger spectra recorded after the first three cycles of CdTe deposition on InP(100) were consistent with the layer-by-layer CdTe growth. It is interesting to note that Cd did not form a UPD deposit on the In-terminated InP(100) surface and only formed Cd clusters at an overpotential. This issue is probably related to the inability of the Cd and In to form a stable surface compound.

Journal Article↗

PubMed2000

Development of terfenadine-pseudoephedrine double-layer tablet dissolution-equivalent to core tablet.

The terfenadine-pseudoephedrine dosage form discussed here is the sustained-release core tablet composed of outer (fast-release) and inner (sustained-release) layers. To develop the double-layer tablet dissolution-equivalent to a core tablet, the fast-release and sustained-release layers were prepared using various disintegrants and polymers, respectively. The layer composed of terfenadine/pseudoephedrine/lactose/cornstarch/sodium bicarbonate/hydroxypropylcellulose (HPC)/sodium lauryl sulfate/microcrystalline cellulose (60/10/90/30/20/1/40/1/293 mg), which gave the fast disintegration time and high dissolved amounts of drugs, was selected as the fast-release layer. The dissolved amounts of pseudoephedrine from sustained-release layers increased more with a smaller ratio of ethylcellulose and hydroxypropylmethylcellulose (HPMC). Dissolution mechanism analysis showed the release of pseudoephedrine was proportional to the square root of time, indicating that drug might be released from the layers by Fickian diffusion. The layer composed of pseudoephedrine/ethylcellulose/HPMC (110/30/155 mg), which had similar dissolution amounts of pseudoephedrine as the inner layer of a core tablet, was selected as the sustained-release layer. Furthermore, the dissolved amounts of drugs from the core and double-layer tablets had deviations of less than 5% against the average dissolved amounts of drugs at each time. There was no significant difference between the dissolved amounts of drugs from these tablets at each time in pH 1.2, 4.0, and 6.8 (P > .05). Our results suggest that this double-layer tablet was a dissolution equivalent to the core tablet.

Chemistry, Pharmaceutical↗

PubMed2004

Optical measurements of absorption changes in two-layered diffusive media.

We have used Monte Carlo simulations for a two-layered diffusive medium to investigate the effect of a superficial layer on the measurement of absorption variations from optical diffuse reflectance data processed by using: (a) a multidistance, frequency-domain method based on diffusion theory for a semi-infinite homogeneous medium; (b) a differential-pathlength-factor method based on a modified Lambert-Beer law for a homogeneous medium and (c) a two-distance, partial-pathlength method based on a modified Lambert-Beer law for a two-layered medium. Methods (a) and (b) lead to a single value for the absorption variation, whereas method (c) yields absorption variations for each layer. In the simulations, the optical coefficients of the medium were representative of those of biological tissue in the near-infrared. The thickness of the first layer was in the range 0.3-1.4 cm, and the source-detector distances were in the range 1-5 cm, which is typical of near-infrared diffuse reflectance measurements in tissue. The simulations have shown that (1) method (a) is mostly sensitive to absorption changes in the underlying layer, provided that the thickness of the superficial layer is approximately 0.6 cm or less; (2) method (b) is significantly affected by absorption changes in the superficial layer and (3) method (c) yields the absorption changes for both layers with a relatively good accuracy of approximately 4% for the superficial layer and approximately 10% for the underlying layer (provided that the absorption changes are less than 20-30% of the baseline value). We have applied all three methods of data analysis to near-infrared data collected on the forehead of a human subject during electroconvulsive therapy. Our results suggest that the multidistance method (a) and the two-distance partial-pathlength method (c) may better decouple the contributions to the optical signals that originate in deeper tissue (brain) from those that originate in more superficial tissue layers.

Absorption↗

PubMed1999

Intracortical excitation of spiny neurons in layer 4 of cat striate cortex in vitro.

Recordings were made from pairs of neurons in cat striate visual cortex in vitro to study the AMPA-channel-mediated components of intracortical excitatory synaptic connections between layer 4 spiny neurons and between layer 6 and layer 4 spiny neurons. Forty-six of the 72 cells recorded were identified morphologically. They consisted of spiny stellate and pyramidal cells in layer 4, and pyramidal cells in layer 6. Connections between layer 4 excitatory cells involve excitatory postsynaptic potentials (EPSPs) averaging 949 microV, with an average coefficient of variation of 0.21 (n = 30). The synapses operate at very high release probabilities (0.69-0.98). With repetitive stimulation these EPSPs show varying degrees of depression, largely mediated by presynaptic changes in release probability. Four pairs of layer 4 cells were reciprocally connected. The connections from layer 6 to layer 4 involve smaller, more variable EPSPs, with an average amplitude of 214 microV, and average coefficient of variation 0.72 (n = 7). These synapses operate at moderately high release probabilities (0.37-0.56). They show facilitation with repetitive stimulation, mediated largely by presynaptic changes in release probability. One excitatory connection from a layer 4 neuron to a layer 6 pyramidal cell was also detected. Thus, layer 4 spiny neurons receive effective excitation from two intracortical sources that have different synaptic dynamics and are likely to contribute significantly to the temporal properties of these cells in vivo.

Animals↗

PubMed2005

Disruption of layer 4 development alters laminar processing in ferret somatosensory cortex.

Treatment with the anti-mitotic agent methylazoxymethanol (MAM) on embryonic day 33 (E33) in ferrets changes features of somatosensory cortex. These include dramatic reduction of cells in layer 4, and altered distributions of thalamocortical afferent terminations and GABA(A) receptors. To determine the effect of the relative absence of layer 4 on processing of sensory stimuli we used current source-density profiles to assess laminar activity patterns. Nearly synchronous activation occurs across all layers in treated animals, which contrasts with the normal cortical activation pattern of initial sinks in layer 4. This change after MAM treatment is consistent with the absence of layer 4 cells and widespread termination of thalamocortical afferents. Using periodic stimulation at 'flutter' frequency, layer 4 neurons in normal somatosensory cortex fire reproducibly to the stimulus rate; the capacity for entrainment is best for layer 4 and weaker in the extragranular layers. The capacity to encode periodic sensory stimuli is disrupted in MAM-treated somatosensory cortex; after an initial response to the onset of periodic stimuli, neurons in all cortical layers show weak entrainment. Neural responses to sensory drive in E33 MAM-treated cortex are also embedded in levels of neural activity substantially above those in normal somatosensory cortex. Sustained stimulation additionally reveals different capacities in each layer for improved signal-to-noise ratios, with layer 4 neurons in normal animals exhibiting the most improved signaling over time. We conclude that normal thalamic terminations, an intact layer 4 and subsequent intracortical processing are integral to proper encoding of stimulus features.

Animals↗

PubMed1997

Importance of the lipid layer in human tear film stability and evaporation.

PURPOSE: Previous work on rabbits has demonstrated a four-fold increase in tear evaporation when the tear lipid layer is removed. However, in vitro work has suggested that the lipid layer does not play a role in retarding evaporation of the aqueous layer. The importance of the lipid layer in human tear film stability and evaporation was determined in the current study by measurement of these parameters in the same individuals. METHODS: The left eyes of 161 normal and dry eye subjects (72 males, 89 females), with an age range of 13 to 85 years, were examined. Tear evaporation was derived from the vapor pressure gradient measured with a modified Servomed evaporimeter. Lipid layer structure and noninvasive break-up time (NIBUT) were assessed clinically, by specular reflection, with the Keeler Tearscope. Lipid layer structure was categorized into marmoreal (open and closed meshwork), flow, amorphous, and colored fringe (normal and abnormal) patterns. These observed patterns reflect lipid layer thickness. Ambient temperature and relative humidity remained fairly constant throughout the experiment. RESULTS: Tear evaporation rate was found to vary significantly with different lipid layer patterns. Eyes with no visible lipid layer, or exhibiting an abnormal colored fringe pattern (with clumping of lipid amidst areas of little or no lipid cover), demonstrated a significantly higher rate of evaporation of the tear film (p < 0.001). There were no significant differences amongst the remainder of the patterns. The NIBUT was also found to vary significantly with lipid layer pattern (p < 0.001), with the absent or abnormal colored fringe lipid patterns exhibiting the poorest stability. CONCLUSIONS: Where the human lipid layer is absent, or is not confluent, and the tear film is unstable, tear evaporation is increased four-fold. However, where there is a stable, intact lipid layer, regardless of lipid thickness, tear evaporation is retarded.

Adolescent↗

PubMed2000

Upper bounds on convective heat transport in a rotating fluid layer of infinite prandtl number: case of intermediate taylor numbers

By means of the Howard-Busse method of the optimum theory of turbulence we obtain upper bounds on the convective heat transport in a heated from below layer of fluid of infinite Prandtl number rotating with a constant angular velocity about the vertical axis. We consider the region of intermediate Taylor numbers: alpha41<<Ta<<alpha61 where alpha(1) is the wave number connected to the 1-alpha-solution of the variational problem. The studied optimum fields possess a three-layer or four-layer structure: in addition to the internal, intermediate, and boundary layers, Ekman layers could arise between the intermediate and boundary ones. For the discussed interval of Taylor numbers the intermediate layers do not expand in the direction of the internal layers. We present an asymptotic theory for the case of the fluid layer with rigid lower boundary and stress-free upper boundary. We use an improved solution of the Euler-Lagrange equations of the variational problem for the intermediate sublayer of the optimum field. This solution leads also to correction of the thicknesses of the boundary layers and to lowering of the upper bounds on the convective heat transport for the cases of fluid layer with stress-free or with rigid boundaries. Thus the known upper bounds for these cases can be treated as upper bounds on the upper bounds on the convective heat transport. For the case of the fluid layer with stress-free boundaries the four-layer optimum fields leads to bounds on the convective heat transport which change from R(1/3) at the lower boundary of their interval of validity to values slightly large than R(2/7) near the upper boundary of the interval of validity. Finally we discuss the area of application of the obtained bounds with respect to the Taylor number Ta and Rayleigh number R.

Journal Article↗

PubMed2006

Suppression of instability in liquid flow down an inclined plane by a deformable solid layer.

The linear stability of a liquid layer flowing down an inclined plane lined with a deformable, viscoelastic solid layer is analyzed in order to determine the effect of the elastohydrodynamic coupling between the liquid flow and solid deformation on the free-surface instability in the liquid layer. The stability of this two-layer system is characterized by two qualitatively different interfacial instability modes: In the absence of the deformable solid layer, the free surface of the liquid film undergoes a long-wave instability due to fluid inertia. With the presence of the deformable solid layer, the interface between the fluid and the solid undergoes a finite-wavelength instability when the deformable solid becomes sufficiently soft. The effect of the solid layer deformability on the free-surface instability of the liquid film flow is analyzed using a long-wave asymptotic analysis. The asymptotic results show that for a fixed Reynolds number and inclination angle, the free-surface instability is completely suppressed in the long-wave limit when the nondimensional (inverse) solid elasticity parameter Gamma=Va eta/(GR)increases beyond a critical value. Here, Va is the average velocity of the liquid film flow, eta is the viscosity of the liquid, G is the shear modulus of the solid layer, and is R the thickness of the liquid layer. The predictions of the asymptotic analysis are verified and extended to finite wavelengths using a numerical solution, and this indicates that the suppression of the free-surface instability indeed continues to finite wavelength disturbances. Further increase of Gamma is found to have two consequences: first, the interface between the liquid and the deformable solid layer could become unstable at finite wavelengths; second, the free-surface interfacial mode could also become unstable at finite wavelengths due to an increase in solid layer deformability. However, our numerical results demonstrate that, for a given average velocity, there exists a sufficient window in the value of shear modulus G where both the unstable modes are absent at all wavelengths. Our study therefore suggests that soft solid layer coatings could potentially provide a passive method of suppressing free-surface instabilities in liquid film flows.

Journal Article↗

PubMed1995

Epileptiform activity in the guinea-pig neocortical slice spreads preferentially along supragranular layers--recordings with voltage-sensitive dyes.

The spread of epileptiform activity was monitored in guinea-pig neocortical slices by the use of a voltage-sensitive dye (RH795) and a fast optical recording technique. Epileptiform activity induced by bicuculline methiodide (10-20 microM) and single-pulse stimulation spread from the stimulation site in layer I or in the white matter across most of the slice. Different lesions were made in the slice in order to specify the neuronal connections used for spread in the horizontal direction. In the slice, intracortical connections are necessary for the spread of epileptiform activity, as shown by vertical cuts through all cortical layers but sparing the white matter. Horizontal connections were interrupted by cuts parallel to the axis of pyramidal neurons through either supragranular or infragranular layers. Vertical connections were interrupted by cuts perpendicular to the axis of pyramidal neurons separating supragranular and infragranular layers. Spread of epileptiform activity in the horizontal direction was not hindered by horizontal cuts. Vertical cuts through infragranular layers also did not hinder the spread of epileptiform activity. In contrast, vertical cuts through supragranular layers either abolished completely (nine slices) or delayed significantly (ten slices) the spread of epileptiform activity. The mean delay at the supragranular lesion was 44 ms in layer III and 30 ms in layer V; at the infragranular lesion the mean delay was 2 ms in layer III and 6 ms in layer V. Also, with horizontal cuts, in three out of five slices the velocity of spread was significantly lower in infragranular as compared to supragranular layers. It is concluded that both supra- and infragranular layers if isolated possess the ability to initiate and propagate epileptiform activity independently. However, in the intact slice the influence of the supragranular networks on initiation and propagation of epileptiform activity appears to dominate.

Animals↗

PubMed2006

Identification of the retrotrigonal layer as a key anatomical landmark during robotically assisted radical prostatectomy.

OBJECTIVE: To define the gross and microscopic anatomy, and the surgical significance, of the newly described retrotrigonal layer of the bladder neck, as an aid during robotic radical prostatectomy (RRP). MATERIALS AND METHODS: The data for this study were obtained from five fresh cadaveric dissections and 100 consecutive RRPs. Five male cadavers with no previous pelvic or urethral surgery were dissected to expose the posterior bladder neck and identify the retrotrigonal layer. This same layer was reproducibly identified in all 100 RRPs. The gross appearance of this layer, and its relationship to neighbouring structures, was documented using still photographs and intraoperative video. The microscopic composition of this layer was assessed using routine haematoxylin and eosin stains, and special staining for smooth muscle (desmin), neural tissue (S-100), and elastin. RESULTS: Grossly, the retrotrigonal layer is a well-defined midline strip located posterior to the bladder neck, that extends from the trigone (superiorly) to the base of the prostate (inferiorly). Microscopically, this layer was predominantly composed of smooth muscle with intervening connective tissue. The retrotrigonal layer serves as a key anatomical landmark to facilitate posterior prostatic dissection, particularly in men with large prostates, prominent median lobes, or previous transurethral prostatic surgery. Further, in our practice, this layer marks the posterior limit of dissection in which electrocautery is still used. Finally, the retrotrigonal layer serves to buttress the posterior layer of the urethrovesical anastomosis. CONCLUSION: The retrotrigonal layer is a key landmark during the posterior bladder neck dissection in the antegrade technique of RRP.

Adult↗

PubMed1983

Electrical activities of the muscle layers of the canine colon.

The spontaneous electrical and mechanical activities of the circular and longitudinal muscle layers of the canine colon were studied. The smooth muscle cells of the circular muscle layer exhibited regular, omni-present myogenic slow-wave activity at a frequency ranging from 4 to 7 c/min. With intracellular micro-electrodes, the slow-wave amplitude was 21-38 mV and its duration 3-6 sec. The 'resting' membrane potential was -60 to -76 mV. Some slow waves had superimposed spike bursts on their peak depolarizations and only these were associated with phasic contractions. It is concluded that they serve a pace-maker function similar to their counterpart in the small intestine. The longitudinal muscle layer exhibited periods of electrical activity alternating with periods of electrical quiescence. During the activity periods electrical oscillations occurred at a frequency of 13-35 c/min with spikes on top of them. Each electrical activity period was associated with a prolonged 'tonic' contraction. The duration of these periods was 30-120 sec and their frequency 0.4-1.1 period/min. This activity is similar to that recorded from the longitudinal muscle of the guinea-pig caecum despite the anatomical differences. The electrical activity periods of the longitudinal muscle appeared to require an excitatory input (stretch and/or acetylcholine release). Provided the strips were not excessively stretched, atropine abolished all electrical and motor activity. Stretching prolonged the electrical activity periods until they eventually fused together and the muscle developed maintained tone. Simultaneously recording from both layers showed that, although electrotonic spread between the two layers is probably insignificant, the activity of the two layers was co-ordinated. Only those slow waves of the circular layer that occurred during the electrical activity periods of the longitudinal layer had superimposed spikes. It is suggested that this co-ordination may indicate that the two muscle layers may be commanded by a common input from periodically active, cholinergic intramural neurones. It is proposed that the complex patterns of colonic electrical and motor activities may be explained as consisting of two major components: one arising from the longitudinal (long spike bursts, high-frequency oscillations and tonic contractions) and the other from the circular layer (slow waves, short spike and phasic contractions). Simultaneous electrical records from the two muscle layers and the mucosa failed to show a consistent relationship between the mucosal record and the activity of either layer. Caution should be exercised in the interpretation of intraluminally derived electrical recordings.

Action Potentials↗

PubMed2006

The modeling of the radiation and response green's function of a fluid-loaded cylindrical shell with an external compliant layer.

Scattering and radiation of acoustic waves from a fluid-loaded cylindrical shell with an external compliant layer are of interest. The compliant layer can be modeled by a normally reacting impedance layer, which has the advantage that complex compliant layer geometries, such as partial compliant layers, can be considered. A question may, however, arise as to the accuracy of this approach. A more rigorous approach is to model the elastic shell and compliant layer using a multilayer shell theory, which has the disadvantage that it cannot be extended to consider partial layers. In this paper scattering results from the normally reacting compliant layer model are compared to those from the multilayer shell model to show that the two approaches produce similar results, except for thickness resonances of the compliant layer. Having established the consistency between the two approaches, results for the far-field acoustic radiation as a function of frequency and radiation angle for a fluid-loaded shell with an external compliant layer excited by an internal ring force are obtained using the normally reacting impedance layer model. These results clearly show the reduction in the far field radiation due to the presence of the compliant layer.

Journal Article↗

PubMed1998

The S-layer proteins of two Bacillus stearothermophilus wild-type strains are bound via their N-terminal region to a secondary cell wall polymer of identical chemical composition.

Two Bacillus stearothermophilus wild-type strains were investigated regarding a common recognition and binding mechanism between the S-layer protein and the underlying cell envelope layer. The S-layer protein from B. stearothermophilus PV72/p6 has a molecular weight of 130,000 and assembles into a hexagonally ordered lattice. The S-layer from B. stearothermophilus ATCC 12980 shows oblique lattice symmetry and is composed of subunits with a molecular weight of 122,000. Immunoblotting, peptide mapping, N-terminal sequencing of the whole S-layer protein from B. stearothermophilus ATCC 12980 and of proteolytic cleavage fragments, and comparison with the S-layer protein from B. stearothermophilus PV72/p6 revealed that the two S-layer proteins have identical N-terminal regions but no other extended structurally homologous domains. In contrast to the heterogeneity observed for the S-layer proteins, the secondary cell wall polymer isolated from peptidoglycan-containing sacculi of the different strains showed identical chemical compositions and comparable molecular weights. The S-layer proteins could bind and recrystallize into the appropriate lattice type on native peptidoglycan-containing sacculi from both organisms but not on those extracted with hydrofluoric acid, leading to peptidoglycan of the A1gamma chemotype. Affinity studies showed that only proteolytic cleavage fragments possessing the complete N terminus of the mature S-layer proteins recognized native peptidoglycan-containing sacculi as binding sites or could associate with the isolated secondary cell wall polymer, while proteolytic cleavage fragments missing the N-terminal region remained unbound. From the results obtained in this study, it can be concluded that S-layer proteins from B. stearothermophilus wild-type strains possess an identical N-terminal region which is responsible for anchoring the S-layer subunits to a secondary cell wall polymer of identical chemical composition.

Amino Acid Sequence↗

PubMed2006

The suprabasal layer of corneal epithelial cells represents the major barrier site to the passive movement of small molecules and trafficking leukocytes.

AIMS: To investigate the site of barrier function to the passive diffusion of a small molecule (phalloidin) in the corneal epithelium in the mouse. METHODS: Penetration of phalloidin (molecular weight 1115 daltons) into the cornea was evaluated by studying fluorescent binding of phalloidin to actin in tissue sections, in whole mount preparations, and in the fixed intact globe by confocal microscopy. In addition, the location of tight junction proteins in the individual layers of the corneal epithelium was determined by immunohistochemistry. RESULTS: Phalloidin staining of corneal sections was positive in all corneal layers in tissue sections and in all layers of the corneal epithelium except the suprabasal layer in excised fixed whole mounts of the cornea. However, when phalloidin staining was attempted in intact fixed globes, before excision of the cornea for whole mount preparation, only the most superficial layer of cells was stained indicating that phalloidin could not penetrate the tissue beyond the suprabasal epithelial layer. Detergent (Triton X-100) treatment of the excised cornea and the intact fixed globe, allowed penetration of phalloidin into the suprabasal epithelial layer. Tight junction proteins occludin, ZO-1 and claudin were present in most layers of the cornea but while ZO-1 and occludin were distributed in a typical pericellular pattern, claudin seemed to be particularly prominent in the suprabasal layer and appeared only as a discontinuous punctate pericellular pattern in the superficial layer. Intraepithelial leukocytes were detected in the superficial epithelium and the basal epithelium but not in the suprabasal epithelium. CONCLUSION: The suprabasal epithelium cell layer appears to represent the main barrier site to the passage of small molecules and cells in the mouse cornea and this property may be attributable to prominent claudin expression in this layer.

Actins↗

PubMed1993

Visual resolution and sensitivity of single cells in the primary visual cortex (V1) of a nocturnal primate (bush baby): correlations with cortical layers and cytochrome oxidase patterns.

1. This study describes the response properties of V1 cortical cells in a nocturnal primate and examines the receptive field organization of these cells in relationship to anatomically defined layers and cytochrome oxidase (CO) rich blobs and CO poor interblob compartments. Visual resolution and contrast sensitivity are consistent with other physiological and behavioral measures in this species. Comparisons are made with response properties of the same zones in macaque monkey, as well as of area 17 of a distantly related species (cat) that also occupies a nocturnal niche. 2. The responses of single cells to drifting sinusoidal gratings were recorded in V1 (striate cortex) of anesthetized, paralyzed bush babies (Galago crassicaudatus). Cells tended to be grouped with respect to ocular dominance, orientation preference, and direction selectivity. There was a high proportion of monocularly driven cells as in macaque monkey. Only 6% of the cells were nonoriented. These were poorly tuned complex cells and bore no resemblance to nonoriented lateral geniculate nucleus (LGN)-like cells reported in layer IV of macaque monkeys. Unidirectional cells were most frequently encountered in cortical layers that receive input from the magnocellular layers of the LGN. 3. Cells were classified as simple (31%) or complex (69%) according to standard criteria. Simple cells were significantly more narrowly tuned than complex cells for both orientation and spatial frequency. Complex cells had significantly higher average optimal spatial frequencies and spatial frequency cutoffs than simple cells. Contrast sensitivity of simple and complex cells averaged 38 and 34, respectively. Spatial resolution and sensitivity of these cells matches behavioral measures in bush baby. The spatial and temporal resolution of bush baby cells are similar to those of cats, which is likely related to the nocturnal niche of both species. 4. Cells in supragranular (I-III) and infragranular (V, VI) layers differed significantly in their response characteristics. The cells in the supragranular layers had significantly higher contrast sensitivity than did the cells in the infragranular layers. Cells in the supragranular layers likewise had higher temporal frequency cutoffs, significantly lower optimal spatial frequencies, lower spatial frequency cutoffs, and tighter orientation tuning than did cells in the infragranular layers. 5. Properties of cells in individual layers and CO blob and interblob compartments also showed differentiation. Layer III had the narrowest orientation and spatial frequency tuning with the tightest tuning in layer IIIC (IVB).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1996

Layer I neurons of the rat neocortex. II. Voltage-dependent outward currents.

1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to study voltage-dependent potassium currents in layer 1 neurons and layer II/III pyramidal cells. 2. In the presence of tetrodotoxin, step depolarizations evoked an outward current. This current had a complex waveform and appeared to be a composite of early and late components. The early peak of the composite K+ outward current was larger in layer I neurons. 3. In both layer I and pyramidal cells, the composite outward K+ current could be separated into two components based on kinetic and pharmacological properties. The early component was termed I(A) because it was a transient outward current activating rapidly and then decaying. I(A) was more sensitive to blocking by 4-aminopyridine (4-AP) than tetraethylammonium (TEA). The second component, termed the delayed rectifier or I(DR), activated relatively slowly and did not decay significantly during a 200-ms test pulse. I(DR) was insensitive to blocking by 4-AP at concentrations up to 4 mM and blocked by > 60% by 40-60 mM TEA. 4. I(A) kinetics were examined in the presence of 40-60 mM TEA. Under these conditions, I(A) began to activate between -40 and -30 mV. Half-maximal activation occurred around 0 mV. In both layer I and pyramidal cells, the half-inactivation potential (Vh-inact) was around or more positive than -50 mV. At -60 mV, > 70% of I(A) conductance was available. I(A) decayed along a single exponential time course with a time constant of approximately 15 ms. This decay showed little voltage dependence. 5. In both layer I and pyramidal cells, I(DR) was studied in the presence of 4 mM 4-AP to block I(A) and in saline containing 0.2 mM Ca2+ and 3.6 mM Mg2+ to reduce contributions from Ca2+-dependent K+ currents. Under these conditions, I(DR) began to activate at -35 to -25 mV with Vh-act of 3.6 +/- 4.5 mV (mean +/- SD). The 10-90% rise time of I(DR) was 15 ms at 30 mV. At 2.2 ms after the onset of the command potential to +30 mV, I(DR) could reach a significant amplitude (approximately 1.5 nA in layer I neurons and 2.2 nA in pyramidal cells depending on the cell size). When long test pulses (> or = 1,000 ms) were used, a decay time constant approximately 800 ms at +40 mV was observed. In both layer I and pyramidal cells, steady state inactivation of I(DR) was minimal. 6. These results indicate that I(A) and I(DR) are the two major hyperpolarizing currents in layer I and pyramidal cells. The kinetics and pharmacological properties of I(A) and I(DR) were not significantly different in fast-spiking layer I neurons and regular-spiking layer II/III pyramidal cells. The relatively positive activation threshold (more than or equal to -40 mV) of both I(A) and I(DR) suggest that they do not play a role in neuronal behavior below action potential (AP) threshold and that their properties are more suitable to repolarize AP. The greater density of I(A) in layer I neurons appears responsible for fast spike generation.

Action Potentials↗