Distribution of inhibitory synapses on the somata of pyramidal neurons in cat motor cortex

The mechanisms by which cortical neurons perform spatial and temporal integration of synaptic inputs are dependent, in large part, on the numbers, types, and distributions of their synapses. To further our understanding of these integrative mechanisms, we examined the distribution of synapses on identified classes of cortical neurons. Pyramidal cells in the cat motor cortex projecting either to the ipsilateral somatosensory cortex or to the spinal cord were labeled by the retrograde transport of horseradish peroxidase. Entire soma of selected corticocortical and corticospinal cells were examined using serial-section electron microscopy. The profiles of these somata and the synapses formed with each of these profiles were reconstructed from each thin section with a computer-aided morphometry system. All somatic synapses were of the symmetrical, presumably inhibitory type. For both cell types, these synapses were not homogeneously distributed over the somatic membrane, but were clustered at several discrete zones. The number and density of synapses on the somata of different corticocortical and corticospinal neurons were not significantly different. However, the density of these synapses was inversely correlated with the size of their postsynaptic somata. We discuss the significance of these findings to the integrative properties of cortical neurons.     

Perforated synapses and plasticity

Against a background of existing models relating perforated synapses to synaptic plasticity, the numerical density and frequency of perforated synapses in rat neocortex have been assessed from 1 d to 22 mo of age using the disector procedure, and changes in their morphology were assessed using 3-D computer reconstructions. The data point toward perforated and nonperforated synapses being separate synaptic populations from early in development, and with perforated synapses playing a part in the maintenance of neuronal postsynaptic density surface area from mid-adulthood onwards. This suggests that they play a crucial role in synaptic plasticity, although its nature may be different from that postulated by most recent workers.     

Fluctuation analysis of short-circuit current in a warm-blooded sodium-retaining epithelium: Site current,density, and interaction with triamterene

Summary Density and conductance of the Na-site in hen coprodeum were studied by employing fluctuation analysis of shortcircuit current at sodium concentrations from 26 to 130mm. Fluctuations of current in the frequency range 2–800 Hz were induced by triamterene, a reversible blocker of conducting epithelial Na-sites. At 130mm Na the site density was 5.8±1.0 m^–2 and the site conductance was 4 pS. This conductance is equal to that of the frog skin (W. Van Driessche and B. Lindemann, 1979,Nature (London) 282:519–520). Extrapolation of site density to zero sodium renders a total of 38±28 sites m^–2, which is compared with other estimates for the coprodeum. The site-triamterene association and dissociation constants were 9.5±0.4 rad sec^–1 m ^–1 and 255±20 rad sec^–1 and they were independent of external sodium concentration. An analysis of the affinity constant for triamterene based on the DC-short-circuit current was found to be unrelated to the external sodium concentration and identical to that obtained from fluctuation analysis indicating a noncompetitive interaction between sodium and triamterene. Due to the oxygen demand of the epithelium we have developed an experimental method using short data processing times. A new analytical approach using integration of the power density spectrum proved necessary because of low signal-to-noise ratios.     

Suprachiasmatic nucleus: Numbers of synaptic appositions and various types of synapses

Summary The suprachiasmatic nucleus (SCN) of male rats was estimated to contain 16×10⁶ synaptic appositions (unilaterally) or 250×10⁶ appositions in 1 mm³ tissue of the nucleus with an average of 1404 appositions per neuron. There are significantly fewer synaptic appositions in the suprachiasmatic nucleus of female rats (15×10⁶ per SCN or 236×10⁶ in 1 mm³ tissue of SCN with 1264 appositions per neuron on an average). Additionally, numbers of various types of synapses (axo-somatic, invaginated, dendrodendritic and optic) are estimated.Supported by National Health & Medical Research Council of Australia     

Analysis of root images from auger sampling with a fast procedure: a case of application to sugar beet

Manual line-intersect methods for estimating root length are being progressively replaced by faster and more accurate image analysis procedures. These methods even allow the estimation of some more root parameters (e.g., diameter), but still require preliminary labour-intensive operations. Through a task-specific macro function written in a general-purpose image analysis programme (KS 300 – Zeiss), the processing time of root images was greatly reduced with respect to skeletonisation methods by using a high-precision algorithm (Fibrelength). This has been previously proposed by other authors, and estimates length as a function of perimeter and area of the digital image of roots. One-bit binary images were acquired, aiming at large savings in computer memory, and automatic discrimination of roots against extraneous objects based on their elongation index (perimeter²/area), was performed successfully. Of four tested spatial resolutions (2.9, 5.9, 8.8, 11.8 pixel mm^–1), in clean samples good accuracy in root length estimation was achieved at 11.8 pixel mm^–1, up to a root density of 5 cm cm^–2 on the scanner bed. This resolution is theoretically suitable for representing roots at least 85 m wide. When dealing with uncleaned samples, a thick layer of water was useful in speeding up spreading of roots on the scanner bed and avoiding underestimation of their length due to overlaps with organic debris. A set of fibrous root samples of sugar beet (Beta vulgaris var. saccharifera L.) collected at harvest over two years at Legnaro (NE Italy) was analysed by applying the above procedure. Fertilisation with 100 kg ha^–1 of nitrogen led to higher RLD (root length density in soil) in shallow layers with respect to unfertilised controls, whereas thicker roots were found deeper than 80 cm of soil without nitrogen.     

Comparative suitability of binocular observation, burrow counting and excavation for the quantification of the mangrove fiddler crab Uca annulipes (H. Milne Edwards)

Insect emergence from three different sections of a small mountain brook in Germany was studied: a free-flowing section (`brook'), a section impounded by beavers (`beaver pond') and a beaver dam. The three sections had very different faunas with that of the dam being more similar to that of the brook than that of the pond. The heterogeneity of the fauna was greatest in the dam and lowest in the pond. Median emergence density in the dam was 443 specimens m^–2 d^–1. This was 3.2 times higher than median emergence density in the brook section and 5.5 times higher than median emergence density in the pond section. In particular, the density of emerging Trichoptera (especially filter-feeding species) was increased. Mean annual number of emerging EPTC species was 32.3/trap (779 cm² ground area) in the dam, 18.4/trap in the brook and 11.0/trap in the pond. Emerging biomass was highest in the dam (18.0 – 26.7 g m^–2 yr^–1; 5.0 times higher than in the free flowing section and 5.4 times higher than in the pond section). The composition of a recently broken beaver dam is described by measuring all pieces of wood used in building the dam and analysing sediment samples. The dam consisted of a framework of wood pieces almost entirely of a diameter <5 cm, in which cavities are partly filled with fine sediment (mean organic content = 20%). The `inner surface' of the dam was 2.09 times higher than the dam's ground area and 1.63 times higher than the dam's surface. Beavers, therefore, significantly increase the heterogeneity of faunal composition and productivity of small brooks.     

Phytoplankton assemblage structure and dynamics as indicator [-2pt] of the recent trophic and biological evolution of the western basin of Lake Como (N. Italy)

We studied the phytoplankton assemblage of the western basin of Lake Como (Northern Italy) during 1997. The phytoplankton assemblage was composed of 65 taxa, belonging to six taxonomic groups. Chlorophyta were represented by the highest number of taxa (28) followed by Bacillariophyceae (17), Cyanoprokaryota (9), Dinophyceae (6), Chrysophyceae (3) and Cryptophyta (2). The total assemblage density and biomass ranged from 902 ind ml^–1 and 134.5 mm³ m^–3 in February to 58 766 ind ml^–1 and 9360 mm³ m^–3 in October. The density and biomass variation at three stations showed a common pattern, with higher values in the southern part of the basin where TP concentrations were always greater. The phytoplankton succession was analysed by cluster analysis (average linkage clustering) and non-metric multidimensional scaling (NMDS) ordination, both applied to a dissimilarity matrix obtained from a calculation of the Bray–Curtis index. In general, the seasonal succession followed a simple pattern, with a clear spring phase. These results are discussed considering the trophic evolution of the lake and its recent colonization by Dreissena polymorpha.     

The kinetics of anion equilibrium exchange across the red blood cell membrane as measured by means of35S thiocyanate

Summary Up to a SCN^– concentration of about 110mm, the concentration dependence of SCN^– equilibrium exchange in human red cell ghosts can be represented by the superimposition of two flux components. One component shows saturation kinetics, the other does not. The saturable component has an activation enthalpy of 105 kJ/mole, exhibits arans acceleration by Cl^– and can be inhibited by H₂DIDS. The nonsaturable component has a much lower activation enthalpy of 33 kJ/mole, is slightly reduced intrans acceleration experiments with Cl^– and insensitive to H₂DIDS but susceptible to inhibition by phloretin. At SCN^– concentrations exceeding 110mm, the saturable component undergoes irreversible self inhibition while the nonsaturable component remains unaltered.The half saturation concentration of the saturable flux component increases with decreasing pH from 3.0mm at pH 7.4 to 13.3mm at pH 6.0. Over this pH range, the maximal flux is only slightly increased from 19×10^–12 to 22×10^–12 moles×cm^–2×sec^–1. The nonsaturable flux component also increases slightly.In accordance with previous observations of Wieth (J. Physiol. (London) 207:563–580, 1970), we find that SCN^– increases K⁺ and Na⁺ permeability. The induced cation-permeability is considerably smaller than the SCN^– exchange and the latter does not show the paradoxical temperature dependence that is known to pertain to the former.     

Frequency response of the electroreceptors (“small pit organs”) of the catfish,Ictalurus nebulosus LeS

Summary By electrophysiologically recording the spikes from the ramus lateralis ventralis nervi vagi of the catfishIctalurus nebulosus LeS., we determined the frequency response of the electroreceptors (small pit organs) in unanaesthetized, freely swimming specimens. The response frequency domain we were able to measure ranged from 0,03 to 25 Hz. We found a peak of greatest sensitivity at about 3 to 7 Hz (Fi. 5), using a stimulus with a current density of 5 × 10^–4A/mm² in water with a specific resistance of 20 Ohm. m. The threshold current density of the small pit organs in a homogeneous sinusoidally varying electric field (3 Hz) proved to be 3 × 10^–5 A/mm².

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Zusammenfassung Durch Ableitung von Nervenimpulsen vom ramus lateralis ventralis nervi vagi beim Zwergwels,Ictalurus nebulosus LeS., versuchten wir den Frequenzbereich der Electroreceptoren (small pit organs) bei frei herumschwimmenden, nicht narkotisierten Tieren zu bestimmen. Der wirksame Frequenzbereich, in dem Bestimmungen vorgenommen werden konnten, erstreckte sich von 0,03 bis 25 Hz. Die Empfindlichkeitskurve der small pit organs erreichte einen Gipfel bei Frequenzen von 3–7 Hz (Fig. 5). Die Reizstromdichte betrug dabei 5 × 10^–4A/mm²; der spezifische Wasserwiderstand war 20 Ohm. m. Es stellte sich heraus, daß die Schwellenstromdichte der small pit organs im homogenen elektrischen Wechselstromfeld (3 Hz) 3 × 10^–5 A/mm² betrug.

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We wish to thank Prof. S. Dijkgraaf for his stimulating interest, and Mr. W. J. G. Loos for making the drawings.     

Multiple conductance levels of the dihydropyridine-sensitive calcium channel in GH3 cells

Summary Calcium channels in GH₃ cells exhibit at least five conductance levels when examined in cell-attached or outside-out patches. These channels resemble the high threshold Ca²⁺ current in their range of activation and inactivation, and in their sensitivity to dihydropyridines (DHP). Mean open times for the five levels were brief (<1 msec) in control solutions but increased in the presence of BAY K 8644. In 100mm Ba²⁺ and BAY K 8644, the five predominant slope conductances were 8–9, 12–13, 16–18, 23–24, and 28 pS. The present study is the first report of multiple levels of the DHP-sensitive Ca²⁺ channel occurring with high frequency in native membranes. The range of conductance levels that we observed encompasses the range of conductances found for two other different types of Ca²⁺ channels and indicates that unit conductance should be used with caution as a distinguishing characteristic for identification of different channel types.     

Involvement of afadin in the formation and remodeling of synapses in the hippocampus

In the hippocampus, synapses are formed between mossy fiber terminals and CA3 pyramidal cell dendrites and comprise highly developed synaptic junctions (SJs) and puncta adherentia junctions (PAJs). Dynamic remodeling of synapses in the hippocampus is implicated in learning and memory. Components of both the nectin-afadin and cadherin-catenin cell adhesion systems exclusively accumulate at PAJs. We investigated the role of afadin at synapses in mice in which the afadin gene was conditionally inactivated in hippocampal neurons. In these mutant mice, the signals for not only nectins, but also N-cadherin and β-catenin, were hardly detected in the CA3 area, in addition to loss of the signal for afadin, resulting in disruption of PAJs. Ultrastructural analysis revealed an increase in the number of perforated synapses, suggesting the instability of SJs. These results indicate that afadin is involved not only in the assembly of nectins and cadherins at synapses, but also in synaptic remodeling.     

Effects of sensory deprivation on the development of asymmetrical synapses in mouse barrels

Alterations in the numerical density and structure of asymmetrical synapses were examined in thin sections through barrel D4 in six CD/1 mice, including three controls and three sensory deprived animals. Sensory deprivation was effected by once daily trimming of all large mystacial vibrissae on the contralateral side of the snout from P0. The mice were perfuse-fixed at P20, several days following the termination of rapid synaptic growth during barrel development (White et al. , Somatosens Mot Res 14 : 34-55, 1997). Cerebral hemispheres contralateral to the deprived side were osmicated, sectioned at 40 mum and embedded in plastic for thin sectioning. Sterio's ( J Microsc 134 : 127-136, 1984) procedure combined with serial thin section analysis (Braendgaard and Gundersen, J Neurosci Meth 18 : 39-78, 1986), was applied blindly to systematic random samples of neuropil in barrel hollows and septa. No significant difference in the numerical density, estimated total number, or in the proportion of perforated postsynaptic densities was observed. However, a significant decrease in the diameters of asymmetrical postsynaptic densities was observed in hollow (P < 0.05) and septal (P < 0.05) neuropil of deprived animals. These results demonstrate a significant morphological alteration in asymmetrical synapses of a type consistent with a reduction in synaptic activity consequent to sensory deprivation.     

Distribution of microglia and astrocytes in different regions of the normal adult rat brain

The distribution of glial cells (microglia and astrocytes) in different regions of normal adult rat brain was studied using immunohistochemical techniques and computer analysis. Antibodies against lipocortin 1 (LC1) and phosphotyrosine (PT), as well as an isolectin, GSA B4 (GSA), were used for identification of microglial units, while antibodies against protein S100β allowed us to identify astrocytes. If LC1 was used as a marker, more microglial cells were detected than with the use of PT or GSA. The highest density of LC1-positive microglial cells (on average, 130±5 cells/mm² of the brain section area) was found in the neostriatum, while the lowest density (51±4 cells/mm²) was observed in the medulla oblongata. In general, the density of an LC1-positive microglial population was higher in the forebrain and lower in the midbrain, and the smallest number of these cells was detected in the brainstem and cerebellum. The number of astrocytes was, on average, 2–3 times as large as the number of microglial cells. High density of astrocytes, was found in the hypothalamus and hippocampus (more than 260 cells/mm²); they were more, numerous in the white matter than in the gray matter. Lower densities of this type cells were observed in the cerebral cortex, neostriatum, midbrain, medulla oblongata, and cerebellum (less than 200 cells/mm²).     

Morphological synergism in root hair length,density, initiation and geometry for phosphorus acquisition in Arabidopsis thaliana: A modeling approach

Low phosphorus availability regulates root hair growth in Arabidopsis by (1) increasing root hair length, (2) increasing root hair density, (3) decreasing the distance between the root tip and the point at which root hairs begin to emerge, and (4) increasing the number of epidermal cell files that bear hairs (trichoblasts). The coordinated regulation of these traits by phosphorus availability prompted us to speculate that they are synergistic, that is, that they have greater adaptive value in combination than they do in isolation. In this study, we explored this concept using a geometric model to evaluate the effect of varying root hair length (short, medium, and long), density (0, 24, 48, 72, 96, and 120 root hairs per mm of root length), tip to first root hair distance (0.5, 1, 2, and 4 mm), and number of trichoblast files (8 vs. 12) on phosphorus acquisition efficiency (PAE) in Arabidopsis. SimRoot, a dynamic three-dimensional geometric model of root growth and architecture, was used to simulate the growth of Arabidopsis roots with contrasting root hair parameters at three values of phosphorus diffusion coefficient (D _e=1×10^–7, 1×10^–8, and 1×10^–9 cm² s^–1) over time (20, 40, and 60 h). Depzone, a program that dynamically models nutrient diffusion to roots, was employed to estimate PAE and competition among root hairs. As D _e decreased from 1×10^–7 to 1×10^–9 cm² s^–1, roots with longer root hairs and higher root hair densities had greater PAE than those with shorter and less dense root hairs. At D _e=1×10^–9 cm² s^–1, the PAE of root hairs at any given density was in the order of long hairs > medium length hairs > short hairs, and the maximum PAE occurred at density = 96 hairs mm^–1 for both long and medium length hairs. This was due to greater competition among root hairs when they were short and dense. Competition over time decreased differences in PAE due to density, but the effect of length was maintained, as there was less competition among long hairs than short hairs. At high D _e(1×10^–7 cm² s^–1), competition among root hairs was greatest among long hairs and lowest among short hairs, and competition increased with increasing root hair densities. This led to a decrease in PAE as root hair length and density increased. PAE was also affected by the tip to first root hair distance. At low D _e values, decreasing tip to first root hair distance increased PAE of long hairs more than that of short hairs, whereas at high D _e values, decreasing tip to first root hair distance increased PAE of root hairs at low density but decreased PAE of long hairs at very high density. Our models confirmed the benefits of increasing root hair density by increasing the number of trichoblast files rather than decreasing the trichoblast length. The combined effects of all four root hair traits on phosphorus acquisition was 371% greater than their additive effects, demonstrating substantial morphological synergy. In conclusion, our data support the hypothesis that the responses of root hairs to low phosphorus availability are synergistic, which may account for their coordinated regulation.     

Short-term stimulation of cellular autophagy by furosemide in the thick ascending limb of Henle's loop in the rat kidney

Summary In an effort to investigate the functional relationship between cell-specific work and intracellular degradative processes, the effect of furosemide on cellular autophagy was investigated in two different portions of the nephron, namely, the thick ascending limb of Henle's loop (TAL), which is a main target of this drug, and the proximal convoluted tubule (PCT) as a reference structure. Eight male adult rats were treated with furosemide (60 mg/kg body weight, s.c.). Eight control animals received physiological saline. 1 to 4 h after the injections the animals were killed by perfusion fixation. Small specimens of kidney tissue from the inner stripe of the outer medulla and from the outer cortex were processed for electron microscopy; they were investigated morphometrically for volume fraction and numerical density of autophagic vacuoles (AVs). A significant increase of both parameters (volume fraction: 0.42 × 10^-4 to 1.09 × 10^-4; numerical density: 4.2 × 10⁵/mm³ to 15.5 × 10⁵/mm³) was seen under the influence of furosemide in TAL cells, whereas PCT cells did not show a significant increase in volume fraction or any increase in numerical density of AVs. These data suggest that the functional unloading of TAL, via blocking of the Na⁺- 2Cl^- — K⁺ co-transport by furosemide, results in adaptative structural unloading, i.e., an increased sequestration of cytoplasmic components into AVs, within a short-time interval.     

Quantitative parameters of synaptic apparatus in the ventral nucleus of the medial geniculate body of the cat

The synaptic apparatus in the ventral nucleus of the medial geniculate body (MGBv) of the cat was examined using electron microscopy and stereological methods, which made it possible to measure the synaptic density. Within 7015 µm² of examined sections, 1586 presynaptic terminal (PST) profiles were found, which corresponds to 226.0·10³ PST per 1 mm² of section surface. The PSP were classified into five groups:RL,RS,F,P, andUT, in accordance with their ultrastructural pattern (dimension of PST profile, dimension and shape of synaptic vesicles, and type of synaptic contact, SC) [18–22]. On the above surface, there were 1012 SC formed by PST of different groups, which corresponds to 144.0·10³ SC per 1 mm² of section surface. TheRL-,RS-,F-,P-, andUT-type PST formed 14.8%, 50.1%, 13.1%, 16.8%, and 5.2% of analyzed SC, respectively. The calculated mean SC numerical density equalled (260.8±54.8)·10⁶ SC per 1 mm³ of fixed MGBv tissue. Among them, 40.2·10⁶ (15.4%) belonged toRL-PST, i.e., to axonal terminals of thecolliculus inferior neurons; 130.2·10⁶ (49.9%) toRS-PST, i.e., mostly to axonal terminals of the auditory cortex neurons; and 33.9·10⁶ (13.0%) toF-PST, i.e., to axons of the GABA-ergic interneurons and neurons of the perigeniculate division of the reticular thalamic nucleus. Group-P PST, i.e., terminal structures of the dendritic arborizations of interneurons, formed 42.7·10⁶ (16.4%) SC per 1 mm³, and 13.8·10⁶ (5.3%) SC belonged toUT-PST, i.e., to terminals of unidentified nature. Among 260.8·10⁶ SC in 1 mm³ of tissue, only 23.8·10⁶ (9% of total number)RL-SC, localized on the relay neurons, are directly involved in the MGBv relay function. All other SC transmit control influences from various structures of the nervous system, and provide adjustment of relay function to the constantly changing environmental conditions and varying status of an orgamism. The mean number of SC, localized on an averaged MGBv relay neuron, was calculated as 9100. Among them, about 1200 SC belong toRL-PST, 5200 SC toRS-PST, 1200 SC toF-PST, 1100 SC toP-PST, and 400 SC toUT-PST.Neirofiziologiya/Neurophysiology, Vol. 27, No. 3, pp. 208–219, May–June, 1995.     

Synaptic apparatus of the feline primary auditory cortex (area Al)

Electron microscope research was conducted on the synaptic apparatus of the feline primary auditory cortex (Al). A total of 2096 profiles of axonal terminals (AT) were found over a total area of 8230 µm² of ultrathin slices at different layers of this cortical layer — an average of 255 profiles per 1000 µm² of the surface area on these slices. The AT profiles occupied about 8.9% of the surface of these cross-sections. It was found that 52% of the AT containing synaptic vesicles formed asymmetrical or symmetrical synaptic contacts (83.9% and 16.1% respectively) and that AT had no contacts which could be considered synaptic junctions on 48% of slices. It was also observed that 45.3% of the AT forming contacts synapsed on spines, 48.5% on dendrites, and 6.2% on neuronal somata. Finally, 95.4% and 4.6% of axo-spinal synapses contained rounded and flattened vesicles respectively; equivalent figures for axodendritic synapses were 79.4% and 20.6% respectively and 19.8 and 80.2% for axosomatic synapses. Calculations revealed an average of 322.8 × 10⁶ AT over 1 mm³ of cat auditory cortex. Organizational aspects of synaptic apparatus at different layers of area A1 were ascertained.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 4, pp. 533–543, July–August, 1990.     

Grid-generated turbulence in a mesocosm experiment

In two mesocosm experiments designed to study the effect of nutrients and turbulence on trophic levels below fish larvae in the ecosystem, turbulence was created by oscillating grids. Utilizing two different frequencies, two levels of turbulence (`high' and `low') were generated in a total of eight mesocosms in both experiments. In each mesocosm a two-layer density structure was created. We succeeded in generating turbulence that mimics turbulence found in natural ecosystems, with turbulent eddies on larger scales than those relevant for organism interactions. Energy dissipation rates () were calculated from the frequency spectrums of the measured turbulent velocities. In the second experiment the average energy dissipation rates in the upper layer were 1.9×10^–7 W kg^–1 and 5.5×10^–8 W kg^–1 in the `high' and `low' enclosures, respectively. Finding `background' turbulence levels between 5.3×10^–9 and 3×10 ^–8 W kg^–1, we suggest that future experiments consider turbulent measurements as a standard variable.     

Comparative studies of fructose 1,6-diphosphate aldolase fromEscherichia coli 518 andLactobacillus casei var.rhamnosus ATCC 7469

A comparative study has been carried out with FDP aldolases fromEscherichia coli 518 andLactobacillus casei ATCC 7469, which had been purified 17.6- and 65-fold, respectively. The aldolase ofL.casei was stable only in the presence of mercaptoethanol, whereas that ofE.coli was strongly inhibited at low (1.0×10^–4 m) and activated at high concentrations (2.0×10^–1 m) of the same compound.p-Chloromercuric benzoic acid inhibited both aldolases, with 40% inhibition at 2×10^–5 m withE.coli aldolase against at 2×10^–4 m withL.casei aldolase. Significant differences were also observed in pH optima and Km values.E.coli aldolase exhibited a maximal activity at pH 9.0 and gave a Km value of 1.76×10^–3 m FDP with strong substrate inhibition above 7×10^–3 m, against pH 6.8–7.0 and a Km of 7.04×10^–3 m FDP forL.casei aldolase. Strong resistance ofL.casei aldolase against inhibition by EDTA, Ca²⁺ and Mn²⁺ was observed compared with complete inhibition at concentrations of 20mm, 40mm and 20mm, respectively, withE. coli aldolase. Polyacrylamide gel electrophoresis did not reveal any differences between the two enzyme preparations.The differences of the properties of FDP aldolases from different bacterial genera are discussed in relation to other Class II aldolases.     

Chronic and endogenous regulation of insulin receptors by catecholamines in adipocytes from patients with a phaeochromocytoma

Insulin binding in adipocytes from patients with a phaeochromocytoma (PH) approached that of the controls (C) at low and higher concentrations of unlabeled insulin. The apparent receptor affinity was unchanged (ED50: PH 0.50×10^–9M and C0.60×10^–9M). Scatchard analysis of the binding data using the negative cooperative model revealed a 46% decrease in the total number of receptors together with no changes in both K^–e (PH 0.55×10⁹M^–1 and C 0.36×10⁹M^–1) and K^–f (PH 0.13×10⁹ M^–1 and C 0.07×10⁹ M^–1). According to the two site model, an altered proportion in the two classes of insulin binding sites was detected. This was accompanied by a catecholamine-desensitization of the adipocytes to the antilipolytic action of insulin. These events could represent a final situation of a chronic and endogeneous regulation by high levels of catecholamines of insulin receptors in human adipose tissue.