Estimation of spatio-morphofunctional characteristics of presynaptic terminals in the lateral zone at the base of the dorsal horn

This article further develops a statistical/stereological model for estimating 3-dimensional population characteristics of cell elements based on analysis of random (2-dimensional) cross section of planes. This model was used to conduct morphometric electron microscope research on presynaptic terminals (PT) at lateral sites at the base of the dorsal horn of the cat spinal cord. Estimated distributions of PT volumes were thus found, as well as areas of PT axolemma with "corrugation" size of synaptic zones (AZ), and estimates of mean numbers of AZ on a 3-dimensional PT and statistical parameters of AZ location in relation to the point at which the axon passes into the synaptic bouton.Dnepropetrovsk State University. A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 6, pp. 803–811, November–December, 1990.     

Stereometric characteristics of ultrastructure of presynaptic terminals in the dorsal horn of the cat spinal cord

Morphometric statistical techniques were used to investigate geometric aspects of 240 axodendritic presynaptic terminals (PT) located in the dorsal horn of the gray matter of the cat spinal cord. Parameters describing the configuration and average spatial radius of the three-dimensional PT (approximated by irregular ellipsoid were estimated on the basis of the morphometry of random PT sections applying aspects of the theory of probability. Lack of any correlation between location of the active zone and the configurational pattern (extent) of terminals was demonstrated. Densitometric distribution of average spatial PT radii was obtained (mean: 0.820 µm). The relatively limited scatter of this distribution would indicate extremely close similarity between parameters of the PT comprising the test group.Dnepropetrovsk University. A. A. Bogomolets Institute of Physiology, Academy of the Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 3, pp. 403–410, May–June, 1989.     

Statistical estimation of the membrane area and numbers of active sites at presynaptic terminals

A study was made of geometrical features of electron micrographic profiles of presynaptic terminals (PT) in the dorsal horn of the cat spinal cord. A statistical approach whereby an area of enclosed membrane may be estimated on the basis of the parameters of its flat random sections is described. An estimation of the mean area of the plasmalemma of the test PT was obtained using this approach. The probability estimate of the mean number of active sites per presynaptic bouton is considered, as well as the possible use of statistical data so obtained for determining characteristics of physical processes occurring at presynapses.Dnepropetrovsk State University. Translated from Neirofiziologiya, Vol. 22, No. 1, pp. 29–36, January–February, 1990.     

Long-Term Posttetanic Changes of Synaptic Transmission and Synapse Morphology of the Pike <Emphasis Type="Italic">Esox lucius</Emphasis> Olfactory Bulb

In the olfactory bulb (OB) of the pike, phenomena of long posttetanic potentiation (LPTP) and long posttetanic depression (LPTD) are reproduced. Probability of induction of LPTP and level of its expression are shown to depend direct on parameters of density of axo-dendritic synapses in the glomerular neuropil and size of its active zones (AZ). In pikes with the high level of these parameters (the synapse density 5.2 per 20 µm², the mean length of AZ cross-section 293 nm), potentiation appear in 95% of cases, while in pikes with low values (the density 3.1, the AZ length 239 nm) in as low as 30%, with LPTD developing rather often instead of LPTP. The LPTP in the phase of its maximal development (60 min after tetanus) corresponds to an increase of the mean length of AZ cross-section of axodendritic synapses by 13.6%, while the LPTD—to its decrease by 16.5% as compared with control. The LPTP induction phase (5–15 min after tetanus) is not associated with any ultrastructural transformations of postsynaptic AZ of axo-dendritic synapses. By the morphometric method, it has been established that LPTP in the pike OB is of the homosynaptic nature, i.e., is due to an enhancement of synaptic efficiency of the excitatory afferent input just subjected to tetanization. On the whole, by using axo-dendritic synapses of the pike OB as an example it was shown that in simple, flat, asymmetric excitatory synapses a steady enhancement of their effectiveness was accompanied by an increase of length of AZ cross-sections, while a reduction of their efficiency—by its decrease.__________Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 41, No. 3, 2005, pp. 285–292.Original Russian Text Copyright © 2005 by Ruzhinskaya, Gdovskii.     

Morphometric analysis of several intracellular events occurring during the vegetative life cycle of the unicellular algaPolytoma papillatum

Summary Quantitative electron microscopy of serial sections was used to study thePolytoma papillatum cell and some of its constituents (nucleus, chondriome, leucoplast) during its vegetative life cycle.The volumes of cells just entering into or passing through mitosis varied considerably and seemed to determine the number of subsequent division processes.Whereas a volumetric balance existed between the cell (100%) and the chondriome (8–9%) during the whole life cycle, there was a correlation between cell and nuclear volumes (8–10%) only during interphase growth and the onset of mitosis. At telophase the nucleus-to-cell-volume ratio was reduced to 2%, but gradually increased during cytokinesis (4.6% at early cytokinesis; 6.5% at late cytokinesis) until it reached the initial value again in newly formed daughter cells. The leucoplast-to-cell-volume ratio (10–26%) varied considerably without any recognizable dependence upon cell cycling.The mean short axis of mitochondrial profiles was proportional to the mean diameter (=thickness) of the mitochondria; the specific surface (outer membrane area per 100 m³ mitochondrial volume), and the surface-to-volume ratio changed rhythmically. Changes in mitochondrial surface-to-volume ratio (Sc/Vc) were apparently correlated with changes in mitochondrial diameter (Dc). This relationship can be approximately described by the function Sc/Vc=4/Dc.Deviations of the surface-to-volume ratios of the nuclei from the surface-to-volume ratios of idealized spheres of equal size, indicating profound changes in nuclear shape, were found mainly during mitosis.Results were compared with those obtained from other morphometric investigations and discussed with regard to their functional meaning.     

Altered processes of vesicle recycling in hippocampal presynapses during modelling of glutamate neurotoxicity

Morphometric electron microscope research was conducted on hippocampal slices from zone CAl in 1-month-old rats after raising extracellular glutamate concentration to 500 µM for 0.5 h and in control animals. Distributions of principal spatial parameters of presynaptic terminals (PT) were obtained using an original statistical-stereological approach. An abrupt increase was demonstrated in numbers of membrane caveolae associated with phases of vesicle recycling (averaging 13 to a single PT), and an increase in mean area of axolemma, as well as minor changes in the volume of test PT. The findings obtained are examined from the aspect of equivalent changes in the physicochemical mechanisms of endo- and exocytosis. A discussion follows of the potential role of the discovered effects in long-term potentiation processes and in those of neurotoxicity in general.Dnepropetrovsk State University. A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 6, pp. 819–826, November–December, 1990.     

Inhibition in the neurons of the somatosensory cortex and the possibility of intracortical propagation of excitation

Extra- and intracellular leads were used to study the reactions of neurons in the pyramidal tract (PT) of the cat brain to antidromic and afferent effects. It was shown that afferent activation of PT neurons proceeds heterogeneously. Three types of PT neurons were identified, successively involved in the impulse response to afferent stimulation. By means of paired stimuli we determined the heterogeneous changes in sensitivity of late reacting PT neurons. It was found that, under certain conditions, the different IPSP evoked by afferent stimulation or PT stimulation do not prevent the appearance of impulse responses to secondary synaptic activation. A conclusion was drawn from these experiments on the localization of the excitatory intracortical terminals on the somas of the PT neurons and on the limited effect of inhibitory processes upon intracortical propagation of the afferent signal reaching the cortex. A functional scheme of intracortical PT neuron links is presented.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the USSR, Kiev. Translated from Neirofiziologiya, Vol. 3, No. 5, pp. 465–473, September–October, 1971.     

Mitochondrial growth and division during the cell cycle in HeLa cells

The growth and division of mitochondria during the cell cycle was investigated by a morphometric analysis of electron micrographs of synchronized HeLa cells. The ratio of total outer membrane contour length to cytoplasmic area did not vary significantly during the cell cycle, implying a continuous growth of the mitochondrial outer membrane. The mean fraction of cytoplasmic area occupied by mitochondrial profiles was likewise found to remain constant, indicating that the increase in total mitochondrial volume per cell occurs continuously during interphase, in such a way that the mitochondrial complement occupies a constant fraction( approximately 10-11(percent)) of the volume of the cytoplasm. The mean area, outer membrane contour length, and axis ratio of the mitochondrial profiles also did not vary appreciably during the cell cycle; furthermore, the close similarity of the frequency distributions of these parameters for the six experimental time-points suggested a stable mitochondrial shape distribution. The constancy of both the mean mitochondrial profile area and the number of mitochondrial profiles per unit of cytoplasmic area was interpreted to indicate the continuous division of mitochondria at the level of the cell population. Furthermore, no evidence was found for the occurrence of synchronous mitochondrial growth and division within individual cells. Thus, it appears that, in HeLa cells, there is no fixed temporal relationship between the growth and division of mitochondria and the events of the cell cycle. A number of statistical methods were developed for the purpose of making numerical estimates of certain three-dimensional cellular and mitochondrial parameters. Mean cellular and cytoplasmic volumes were calculated for the six time-points; both exhibited a nonlinear, approx. twofold increase. A comparison of the axis ratio distributions of the mitochondrial profiles with theoretical distributions expected from random sectioning of bodies of various three-dimensional shapes allowed the derivation of an "average" mitochondrial shape. This, in turn, permitted calculations to be made which expressed the two-dimensional results in three-dimensional terms. Thus, the estimated values for the number of mitochondria per unit of cytoplasmic volume and for the mean mitochondrial volume were found to remain constant during the cell cycle, while the estimated number of mitochondria per cell increase approx. twofold in an essentially continuous manner.     

Changes in the topography of transmitter release in a neuromuscular synapse reflect plastic alterations occurring in active zones

The topography of transmitter release along the motor nerve terminals (NT) was studied on the frogcutaneous pectoris muscle under normal conditions and following denervation. Coordinates of release sites (RS) of transmitter quanta were determined by extracellular recording of postsynaptic signals using three microelectrodes. It was shown that RS form groupings that reflect transmitter release in individual active zones (AZ). The topography of transmitter release in the distal parts of the NT under normal conditions was shown to differ from that observed in the proximal parts. The difference consists in a lower probability of transmitter release in AZ and a higher probability of this process between AZ, as well as in a change of release profile in individual AZ. Similar differences were found following denervation. It is suggested that these properties may reflect plastic reorganization occurring in AZ in the course of remodelling of neuromuscular synapse and its degeneration.Neirofiziologiya/Neurophysiology, Vol. 27, No. 4, pp. 253–260, July–August, 1995.     

Oviposition bioassay responses of Culex tarsalis and Culex quinquefasciatus to neem products containing azadirachtin

Neem (Azadirachta indica A. Juss) products have been shown to exert pesticidal properties against a variety of insect species. In mosquito control programs, such products may have the potential to be used successfully as larvicides. In exploring other advantages of neem products, we studied the oviposition responses of Culex tarsalis Coquillett and Cx. quinquefasciatus Say to two experimental azadirachtin (AZ) formulations, wettable powder Azad ^– WP10 (WP) and emulsifiable concentrate Azad ^– EC4.5 (EC). Gravid Cx. tarsalis exhibited a distinct preference for the neem suspension of the WP, where significantly more egg rafts were collected from the treatment than from the control. The minimum effective AZ concentration for this activity was 0.5 ppm. The aged suspensions from 1–7 days at 0.5 and 1 ppm AZ were more active in eliciting oviposition responses in Cx. tarsalis than the fresh preparations. This activity of the aged suspensions lasted up to 14 and 21 days at 0.5 and 1 ppm AZ, respectively. Negative ovipositional responses were indicated in the tests of the EC vs. Cx. tarsalis, as well as both neem formulations vs. Cx. quinquefasciatus. In the tests of the EC formulation, significantly less gravid females were trapped by oviposition cups in the treatment than in the control, and in the tests of the WP significantly less egg rafts were collected from the treatment than from the control. The minimum effective concentrations for oviposition avoidance activity were 5 ppm AZ for Cx. tarsalis and 10 ppm AZ for Cx. quinquefasciatus, which lasted up to 1 and 4 days for these two species respectively. Neem products potentially used as mosquito larvicides may have many additional benefits in mosquito control programs, the oviposition modification noted in the current studies is one such example.     

Spectrally Resolved Morphometry of the Nucleus in Hepatocytes Stained by Four Histological Methods

A novel concept of spectrally resolved morphometry for histological specimens was developed using light microscopy combined with spectrally resolved imaging. The spectroscopic characteristics of rat hepatocytes stained by Haematoxylin and Eosin, Romanowsky–Giemsa, periodic acid–Schiff and Masson's trichrome were assessed. Light intensity in the range 450–850 nm was recorded from 10000 pixels of nuclear domains of each stained cell and represented as light transmittance spectra and optical density. In order to identify spectral shifts caused by stain– macromolecule interactions, we compared the spectra of individual stain components with those of DNA and bovine serum albumin. Chromatin and interchromatin areas were classified spectrally using a chosen spectral library followed by morphometric calculations of nuclear domains for each staining method. The spectral fingerprints of Masson's trichrome stain distinguished the nucleolus from the rest of the nuclear c hromatin, enabling the demarcation and calculation of the nucleolar area. Spectrally resolved imaging of human hepatocytes stained by Masson's trichrome stain revealed marked differences between the nucleolar area in normal human hepatocytes compared with hepatocellular carcinoma. Masson's trichrome stain also distinguished the nucleolar area in human breast carcinoma cells and keratinocytes.     

Age-related changes in olfactory sensitivity of laboratory mice

Age-related alterations in the electrical response of olfactory epithelium to odorant-induced stimulation and certain morphometric indices were investigated in male and female laboratory mice of strains C57BL/6 (B6) and AKR (AK). It was found that maximum amplitude of response to odorants characterized young and adolescent animals. Ageing is accompanied by a decline in response level in the olfactory epithelium. Age-related distinctions between morphometric characteristics of the olfactory organ such as overall area and depth of the olfactory epithelium were noted.Applied Mathematics and Cybernetics Research Institute, N. I. Lobachevskii University, Gor'kii. Translated from Neirofiziologiya, Vol. 21, No. 6, pp. 723–729, November–December, 1989.     

Quantitative Evaluation of Diffusion and Dynamic Contrast-Enhanced MR in Tumor Parenchyma and Peritumoral Area for Distinction of Brain Tumors

Purpose

To quantitatively evaluate the diagnostic efficiency of parameters from diffusion and dynamic contrast-enhanced MR which based on tumor parenchyma (TP) and peritumoral (PT) area in classification of brain tumors.

Methods

45 patients (male: 23, female: 22; mean age: 46 y) were prospectively recruited and they underwent conventional, DCE-MR and DWI examination. With each tumor, 10–15 regions of interest (ROIs) were manually placed on TP and PT area. ADC and permeability parameters (K^trans, Ve, Kep and iAUC) were calculated and their diagnostic efficiency was assessed.

Results

In TP, all permeability parameters and ADC value could significantly discriminate Low- from High grade gliomas (HGG) (p<0.001); among theses parameters, Ve demonstrated the highest diagnostic power (iAUC: 0.79, cut-off point: 0.15); the most sensitive and specific index for gliomas grading were K^trans (84%) and Kep (89%). While, in PT area, only K^trans could help in gliomas grading (P = 0.009, cut-off point: 0.03 min^-1). Moreover, in TP, mean Ve and iAUC of primary central nervous system lymphoma (PCNSL) and metastases were significantly higher than that in HGG (p<0.003). Further, in PT area, mean K^trans (p≤0.004) could discriminate PCNSL from HGG and ADC (p≤0.003) could differentiate metastases with HGG.

Conclusions

Quantitative ADC and permeability parameters from Diffusion and DCE-MR in TP and PT area, especially DCE-MR, can aid in gliomas grading and brain tumors discrimination. Their combined application is strongly recommended in the differential diagnosis of these tumor entities.     

Autonomic Cardiac Rhythm Regulation and Temperament in Infancy

The specific features of the cardiac rhythm (CR) and temperament were studied in 92 children aged from 15 to 31 months. The average cardiac interval in a recumbent position and its variability (standard deviation) were determined. The parasympathetic tone (PT) was determined by analyzing the CR wave structure according to CR spectrum power within the range of 0.2 to 1.2 Hz. The sympathetic tone was measured by subtracting the PT contribution from the spectrum power within the range of 0 to 0.2 Hz by means of regression. The children's temperament was measured using their parents' questionnaires. Repeated examination showed the stability of the CR characteristics. The PT was shown to correlate with the children's motor activity (0.52), approaching a new subject (0.39), distraction (–0.34), and the age of entering a nursery (–0.35). The results evidenced a relationship between the parasympathetic tone and the behavioral activity during the first years of life and the role of the parasympathetic tone in physiological regulation of approach/avoidance responses and sustained attention.     

The mitochondrial megachannel is the permeability transition pore

Single-channel electrophysiological recordings from rat liver mitoplast membranes showed that the 1.3-nS mitochondrial megachannel was activated by Ca⁺⁺ and inhibited by Mg⁺⁺, Cyclosporin A, and ADP, probably acting at matrix-side sites. These agents are known to modulate the so-called mitochondrial permeability transition pore (Gunter, T. E., and Pfeiffer, D. R. (1990)Am. J. Physiol. 258, C755–C786) in the same manner. Furthermore, the megachannel is unselective, and the minimum pore size calculated from its conductance is in agreement with independent estimates of the minimum size of the permeabilization pore. The results support the tentative identification of the megachannel with the pore believed to be involved in the permeabilization process.Abbreviations used: PT: permeability transition; PTP: permeability transition pore; MMC: mitochondrial megachannel; IMAC: inner membrane anion channel. P_A: permeability of ion A. CSP: Cyclosporin A.     

Leaf features of silver birch (Betula pendula Roth). Variability within and between two populations (uncontaminated vs Pb-contaminated and Zn-contaminated site)

Fourteen morphometric variables of silver birch (Betula pendula Roth) leaves from two sites in the Upper Silesia province were analyzed to establish the pattern of intrapopulational and interpopulational relations. The leaves were collected from ten trees growing on a zinc–lead dump as well as from ten trees in the unpolluted area of Mirów within the same bioclimatic region. Leaf samples were collected from the trees during the vegetative seasons 1999 and 2000. The size and shape of leaves were studied using standard biometric methods. Cluster analysis indicated overall differences between the populations. Both populations differed with respect to almost all the morphometric variables (P<0.05). Most variables of the leaves, collected from single trees (or combined as a total) on the zinc–lead dump showed more variability than those from the unpolluted site. Discriminant function analysis confirmed the angle of the leaf base as the most important variable in the evaluation of interpopulational variability of leaves.     

Transmitter release at the active zone of motor nerve endings

Transmitter release sites were located in the motor nerve ending of the frog cutaneous-pectoris muscle using three extracellular electrodes. Transmitter release sites were found to be grouped in a direction cutting across the nerve ending and reflecting transmitter release and active release zones (AZ). Measurements from these groups showed that most transmitter release takes place at the center of the AZ, declining towards the periphery and to either side of this zone. All AZ were found to take place in spontaneous release with a low extracellular concentration of calcium ions present, compared with only a proportion in evoked release. Advantages of the triple as opposed to the dual micro-electrode technique are analyzed. It was found that transmitter release in spatially isolated AZ at the nerve ending leads to a polymodal distribution pattern of the amplitude of uniquantal signals during extracellular recording. The part played by AZ in transmitter release is discussed.S. V. Kurashov Medical Institute, Ministry of Health of the RSFSR, Moscow. V. I. Ul'yanov-Lenin State University, Kazan'. Translated from Neirofiziologiya, Vol. 22, No. 3, pp. 318–327, May–June, 1990.     

Regional morphological variations within the crayfish eye

Summary The existence of structural asymmetries has been quantitatively demonstrated in the crayfish compound eye. Variations in the size of the rhabdomes and corneal facets, as well as the size and extent of the accessory reflecting pigment cells, have been found. It was determined that the mean rhabdome diameter within a 70° arc in the dorsal quadrant of the retina is 11–19% smaller than the mean rhabdome diameter in the remaining areas of the eye. Also, the extent of the accessory reflecting pigment cells is diminished over an area corresponding generally to the dorsal region of smaller rhabdomes. Corneal facet size and shape vary over the surface of the cornea, with smaller facets occurring in the dorsal region. Both the mean rhabdome diameter and the mean corneal facet area for whole eyes increases linearly in animals ranging in size from 3.9–12 cm. The estimated number of corneal facets, and therefore the number of rhabdomes, increases from an average of 4700 in the 3–6.9 cm size range to about 6000 in 7–12 cm animals. These data indicate that structural asymmetries and various size-related parameters exist in the crayfish eye and should be considered in any quantitative analysis of this structure.Supported by a grant from the National Science Foundation (BNS 80-04587)     

Numerical characteristics of synaptic vesicular apparatus structure in the dorsal horn of the cat spinal cord

Statistical parameters characterizing the structure of the vesicular apparatus of presynaptic endings (PE) were determined from the findings of ultrastructural morphometric analysis of 135 synapses in the dorsal horn of the cat spinal cord. Quantitative estimates of vesicles in the PE (averaging about 470) were obtained, based on stereometric principles. The bimodal pattern of distribution of distances from the center of each vesicle to the nearest portion of the active zone was demonstrated — viewed as the structural correlate of the two-pool model of transmitter storage. The possibility of classifying PE according to the sign of vesicle spatial distribution is discussed as well as the relationship between this distribution and parameters of transmitter mobilization and synaptic release.Dnepropetrovsk State University. A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 5, pp. 597–605, September–October, 1989.