Structure of visual pathways in the nervous system of freshwater pulmonate molluscs

Central nervous system of freshwater pulmonate molluscs Lymnaea stagnalis and Planorbarius corneus was stained using retrograde transport of neurobiotin in the optic tract fibers. In both species, perikarya and fibers of the stained neurons are found in all ganglia except the buccal ones. Afferent fibers of the optic nerve form dense sensory neuropil located in relatively small volume of cerebral ganglia. Typical neuronal groups sending their processes into the optic nerves of ipsilateral and contralateral body halves are described. Among them, neurons of visceral and parietal ganglia innervating both eyes concurrently as well as sending projections into peripheral nerves are revealed. These neurons, supposedly, have a function to integrate sensory signals, which may be a basis for regulation of light sensitivity of retina and functioning of peripheral organs. Bilateral links of the molluscan eye with the pedal ganglia cells and statocysts are found, which is, likely, a structural basis of certain known behavioral patterns related to stimulation of visual inputs in the studied gastropod molluscs.     

Some aspects of the structural organization of the arthropod ganglia

Summary This paper deals with the fine structure of the abdominal ganglia of several species of arthropods belonging to the classes Arachnida, Crustacea, Myriapoda and Insecta. The tissues were fixed in osmium tetroxide and embedded in n-butyl methacrylate or fixed in potasium permanganate and embedded in a mixture of X 133/2097 and Araldite.A comparative study was made in order to discriminate between those structural characteristics of the nervous system appearing only in determined taxonomic groups and those belonging to a fundamental plan common to the whole Phylum. This work covers the morphology of neurons, glial cells, neuropilic nerve fibers and neuronal connections.Most arthropod neurons are pear-shaped with only one prolongation and the nucleus is located in the center of the soma, enveloped by two membranes showing numerous pores. Cisternae of the ER have frequently been observed in continuity with this nuclear envelope. After osmic fixation the nuclear content appears to consist of small dense granules distributed at random in the nucleoplasm. In addition to these small perticles there are, in some species, large chromatin blocks. The use of Permanganate as fixative introduces important changes in the nuclear aspect; most of the nuclei look washed and the nuclear content acquires an homogeneous appearance.The cytoplasm of the neurons contains a complex system of internal membranes consisting of cisternae and tubuli of the ER system, lamellae of the Golgi complex and invaginations of the plasma membrane. In most species the elements of the ER system are distributed at random in the cytoplasm but in the neurons of Bothriurus bonariensis there are parallel aggregations of membranes similar to the Nissl bodies found in vertebrates.It was found in some of the species studied (Armadillidium vulgare and Lithobius Sp.) that the internal membrane system of the nerve cells is mainly represented by Golgi elements while the ER system seems to be poorly developed.Besides the membranous components, the neuronal cytoplasm contains mitochondria, multivesicular bodies and dense granules of neurosecretory material.Neuroglial cells are mainly characterized by their nuclear structure. After the action of osmium tetroxide, glial nuclei show irregular masses of chromatin inmersed in a nucleoplasm of low electron density. In permanganate fixed material these chromatin blocks appear as blank spaces.In the cytoplasm of these cells there are mitochondria, membranes pertaining to the ER system and elements of the Golgi complex but in some of the species studied gliofibrils and granules of pigment were found.Three main types of neuroglial cells have been recognized in an arthropod ganglia. These are: subcapsular glial cells, neuron satellites and nerve fiber satellites.The neuropile occupies the central region of the ganglion and consists of a great number of nerve fibers intermingled with glial processes. The neuropilic n. fibers consistently show profiles of ER membranes and tubuli pertaining to the ER system. In some of these fibers the ER reaches a high degree of development. In Armadillidium there is a special type of n. fiber containing a regular sequence of transversally oriented cisternae. Arthropod fibers sometimes contain thin parallel filaments as well as typical ER elements.Mitochondria, small vesicles and dense granules are commonly found within the neuroplasm of the neuropilic fibers. It is important to note that in arthropods, microvesicles are not restricted to the terminal region of the nerve fibers but that they may also occur all along the fibers.Arthropod neurons are enveloped by a glial insulating capsule and therefore interneuron contacts may only occur at neuropile level. These contacts are of three different morphological types: cross contacts, longitudinal contacts and end-knob contacts. At the level of longitudinal and cross contacts the neuroplasm shows no increase in the number of microvesicles or mitochondria. In the end-knob contacts, on the contrary, large numbers of microvesicles appear concentrated in the pre-synaptic fiber only, and occasionally in both fibers the pre-synaptic and the post-synaptic.It is maintained that funcional interneuron connections may result not only from contacts between fibers containing vesicles, but also between fibers in which vesicles are absent.     

Orientation of intrinsic proteins in photosynthetic membranes. Polarized infrared spectroscopy of chloroplasts and chromatophores

In order to estimate the degree of orientation of the α-helices of intrinsic proteins in photosynthetic membranes, polarized infrared spectroscopy has been used to measure the dichroism of the amide I and amide II absorption bands of air-dried oriented samples of purple membranes, chloroplasts and chromatophores from Rhodopseudomonas sphaeroides. Using purple membrane, in which the orientation of the α-helices is precisely known (Henderson, R. (1977) Annu. Rev. Biophys. Bioeng. 6, 87–109), as a standard to calibrate our measurements and estimating the mosaic spread (extent of orientation) of the membranes from linear dichroism measurements performed in the visible spectral range, it is concluded that in photosynthetic membranes, the α-helices of intrinsic proteins are tilted at less than 40° with respect to the normal to the plane of the membrane.     

OBLIQUELY STRIATED MUSCLE : IV. Sarcoplasmic Reticulum, Contractile Apparatus, and Endomysium of the Body Muscle of a Polychaete, Glycera, in Relation to Its Speed

Body muscle cells of the bloodworm Glycera, a polychaete annelid, were studied by electron microscopy and compared with muscle cells of the more slowly acting nematode Ascaris, which have been described previously. Both muscles are obliquely striated. The predominant type of bloodworm fiber is characterized by a prominent transversely oriented sarcoplasmic reticulum with numerous dyads at the surface of each cell. Thick myofilaments are ~3 µ long and overlap along ~60% of their length in extended fibers and ~80% in shortened fibers. There is virtually no endomysium and very little intracellular skeleton, and the cells are attached by desmosomes to one another rather than to connective tissue. Dense bodies are absent from the fibers and in their place are Z lines, which are truly linear rather than planar. Scattered among the predominant fibers are others, less orderly in arrangement, in which the SR is much less prominent and in which the thick filaments are thicker and longer and overlap to an even smaller degree. It is suggested that physiological differences between bloodworm and Ascaris muscles derive from differences in the proportion of series to parallel linkages between the contractile elements, differences in the amount and disposition of the SR, and differences in the impedance to shear within the myofibrils.     

ORIENTATION AND LOCUS OF TROPIC PHOTORECEPTOR MOLECULES IN SPORES OF BOTRYTIS AND OSMUNDA

Study of the tropic responses of Botrytis cinerea and Osmunda cinnamomea spores to blue light shows the photoreceptor molecules to be highly dichroic and oriented: in Botrytis their axes of maximum absorption lie perpendicular to the nearby cell surface; in Osmunda, parallel. The chief evidence lies in a comparison of their responses to plane polarized light—both germinate parallel to the vibration planes (defined by the axis of vibration of the electric vector and the axis of light propagation)—with those to partial illumination with unpolarized light: Botrytis grows from its brighter part; Osmunda, from its darker. The degree of orientation produced by polarized light corresponds, at high intensities, to that produced by the imposition of such large (about 100 per cent) intensity differences across a cell as to preclude all alternatives to oriented dichroic receptors. The photoreceptors of the Botrytis spore lie within the cell wall's inner half. The chief evidence lies in the component of its tropic responses to polarized light within the vibration plane: germination peaks about 10° off the vibration axis. This deviation arises from focusing which is effective only in the wall's inner half. At high intensities, anomalies appear in Botrytis which are interpreted as "centering," i.e., a tendency toward growth from the center of two or more equally illuminated points of a cell rather than from one of them.     

The conformation and orientation of copper (II)-bleomycin intercalated with DNA

EPR data are used to describe the conformation and identity of the atoms coordinated to Cu(II) in Cu(II)-bleomycin bound to oriented DNA fibers. The fibers were slowly drawn from viscous solutions of Cu(II)-bleomycin-DNA containing one Cu(II)-bleomycin to 200 basepairs. EPR measurements were made at room temperature and 90 K for different orientations of the external magnetic field with respect to the helical axes of the fibers. The g-values (g parallel = 2.21, g perpendicular = 2.04) and the hyperfine constant (A parallel = 175 G) are consistent with values expected for Cu(II) chelated to a square planar array of ligands. In the oriented fibers, the square planar arrays do not all have the same orientations with respect to the fiber axes. At room temperature the chelated ions have rotational freedom in which the normal to the planar array has almost complete freedom of rotation about axes perpendicular to the DNA fiber axes. The normal maintains an angle of 75 degrees with respect to the axis, in the plane of the basepair, about which it rotates. Nine superhyperfine peaks on the high field side of the EPR spectrum were partially resolved. The number and splitting (12 G) of these superhyperfine peaks indicate that four nitrogen atoms are chelated to Cu(II) in a square planar array. These data on Cu(II)-bleomycin bound to DNA give information on the orientation of the metal-containing portion of bleomycin which lies outside to double helix.     

Structure of visual pathways in the nervous system of fresh water pulmonary molluscs

The central nervous system of freshwater pulmonary molluscs Lymnaea stagnalis and Planorbarins corneus was stained by the method of neurobiotin retrograde transport along optic nerve fibers. In the animals of both species, bodies and fibers of stained neurons are found in all ganglia except for the buccal ones. Afferent fibers of the optic nerve form a dense sensor neuropil located in a small volume of cerebral ganglia. Characteristic groups of neurons sending their processes into optic nerves both of ipsi- and of contralateral half of the body are described. Revealed among them are neurons of visceral and parietal ganglia, which simultaneously innervate both eyes as well as give projections into peripheral nerves. It is suggested that these neurons can perform function of integration of sensor signals and, on its base, regulate photosensitivity of retina as well as activity of peripheral organs. There is established the presence of bilateral connections of the mollusc eye with cells of pedal ganglia and statocysts, which seems to be the structural basis of manifestation of the known behavior forms associated with stimulation of visual inputs of the studied gastropod molluscs.     

Natural polymers according to NMR data: cross-relaxation in hydrated collagen macromolecules from two connective tissues

Spin-lattice relaxation and cross-relaxation in oriented and randomly oriented collagen fibers from two connective tissues (15-month-old calf and 8-year-old steer) at a water content of 0.6 g H2O/g dry matter were studied. Collagens were chosen according to different numbers of covalent nonreducible cross-links, which increase during the life of the animal. The spin-lattice relaxation curves for all the collagens after a 180 degree-tau-90 degree pulse sequence were described by two exponential components. The dependences of two components of spin lattice relaxation time and their populations on the length of the 180 degree-pulse were obtained. On the basis of data of Goldman-Shen sequence and the two-phase model, the populations of proton fractions (p(w) and p(c)) as well as the rates of transfer of magnetization between water protons and collagen protons (k(w) and k(c)) were calculated. No significant difference between k(w) (k(c)) in oriented and randomly oriented fibers as well as in fibers with different cross-linking was found. The estimates of the cross-relaxation times for low cross-link collagen and high cross-link one were done. The correlation times of dipole-dipole interactions for both connective tissues were calculated using the cross-relaxation theory.     

The roles of polarity and symmetry in the perceptual grouping of contour fragments

We describe two experiments that investigate the roles of polarity and symmetry in the perceptual grouping of contour fragments. Observers viewed, for one second on each presentation, arrays of oriented, spatial-frequency band-pass, elements, in which a subset of the elements was aligned along a twisting curve. In each of five conditions we measured observers' ability to detect aligned combinations of even- and odd-symmetric elements, of the same and different polarities, against a background of 'noise' elements. As with previous experiments we found that the 'path' could be reliably detected, even when the elements of the path were oriented at angles of up to +/- 60 deg relative to each other. Detection of the path was still possible when the polarity of path elements alternated. However, the probability of detection of the path was raised significantly when the path elements were all of the same polarity. Perceptual grouping of even-symmetric elements was no different to perceptual grouping of odd-symmetric elements. The results provide evidence, that in achieving integration of contour fragments, the visual system uses a process that is to some degree phase selective. We use the results to describe how the visual system may resolve natural contours when they occur against backgrounds that vary over a wide range of intensities. The data presented here have been published in conference-abstract form (Hayes et al., 1993; Field et al., 1997).     

Das anatomische und das physiologische Sehfeld der Ommatidien im Komplexauge von Musca

Summary The sensitivity distribution of visual elements in the front region of the compound eye of Musca is investigated by means of intracellular micropipette recording. This distribution approximates a Gauss-function of 7.7 degree half-value in the horizontal plane. Using a bleaching effect of the Musca compound eye as an indicator, the angular distances between the optical axes of adjacent ommatidia are measured and found to vary between 2.3 and 3.9 degrees. Thus the visual fields of adjacent ommatidia strongly overlap. Based on these findings a calculation reveals that less than 12% of the mean efficient light flux is received from the anatomically determined visual field of the ommatidium. Similar percentages for Calliphora (less than 20%) and Limulus (less than 19%) result from evaluation of data collected by other investigators. — Light entering the visual element from different directions (more than 5° apart) is demonstrated to be 1.2 to 1.4 times more effective than light received from the same direction. — Consequences of the overlap of visual fields of adjacent ommatidia for perception of motions and patterns by the compound eye are discussed.

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Mit Unterstützung durch die Stiftung Volkswagenwerk. Kybernetik, Band 2     

A look into the cock-pit of the fly

Summary An anatomical investigation has been carried out on the third optic ganglion of the fly, Musca domestica. Two systems of giant units, the dendritic arborizations of which are arranged orthogonally relative to each other, dominate the neuropile of this ganglion. The elements of the two systems have been reconstructed using a graphical procedure based on histological sections. One system branches predominantly in the dorsoventral direction, the other one in the anterior-posterior direction. Both systems of the giant units have a twin system composed of elements smaller in diameter and strictly parallel to the main units. The two systems have been termed the Vertical and Horizontal Systems.The elements of the two systems of fibers project into the periesophageal region where they come into contact with other descending elements. Electron microscopic investigations show that the two systems are post-synaptic at the level of the ganglion from which they originate. The horizontal system has been shown to be post and pre-synaptic in nature during its course in the mid-brain and ultimately presynaptic at its endings in the periesophageal ring. The peculiar geometric arrangement of the two anatomical systems of fibers suggests a precise function in relation to the visual world and in particular to the detection of the direction of motion. The accuracy of the structural pattern displayed by the giant units in the lobular plate seems to suggest that this optic ganglion represents the ultimate orderly projection of the external world in the brain of the fly.A short review of the electrophysiological data concerning this ganglion has been tentatively correlated with some behavioral data related to the visual orientation and fixation in insects.     

Optimized Transection: A Prelude to Oriented Sectioning

A technique is described which permits blocks of tissue to be flat-embedded in euhedral plastic castings and then to be transected along a plane so that sections may be cut which are optimally oriented to the internal ultrastructure of the block. In the transection procedure a hollow plastic cylinder is placed on the specimen trimming block. The cylinder's top prescribes a plane to which the tissue block is accurately oriented and clamped at a predetermined level. Two hand files and a burnisher are worked across the cylinder's top to 1) remove extraneous material above the plane of transection, 2) expose the tissue for sectioning and 3) smooth the block face. The clear plastic at the periphery of the exposed tissue is then easily trimmed away with a razor blade. The result is a block face with a flat, reflective surface which may be quickly aligned to the knife on the ultramicrotome. The effort needed to transect, align and face the block is minimal and 1-micron or semithin sections produced will be precisely parallel to, and at, the plane of transection. Dust produced by the transection procedure is easily eliminated from the work area by use of a small disposable vacuum cleaner. The technique of producing optimally oriented light microscope sections, using the transector, is enhanced by application of solvents to the block face which cause it to develop a temporary low relief, exactly matching the structural detail of sections cut from the block face. Areas of interest can be accurately located and isolated on the block face, using only a hand-held razor blade, so that oriented ultrathin sections of important regions can be routinely cut and examined in the electron microscope.     

Plastid apportionment and preprophase microtubule bands in monoplastidic root meristem cells ofIsoetes andSelaginella

Summary Ultrastructural observations on monoplastidic root tip cells ofIsoetes andSelaginella demonstrate two important phenomena associated with preprophasic preparation for mitotic cell division, 1. the preprophase band and 2. precise orientation of the dividing plastid relative to the preprophase band. Both of these phenomena accurately predict the future plane of cell division. The plastid divides in a plane parallel to the spindle and each cell inherits a single plastid which caps the telophase nucleus. When succesive transverse divisions occur, the plastid migrates prior to prophase from a position near an old transverse wall to a lateral position in the cell. The plastid is oriented with its median constriction precisely intersected by the plane of the preprophase band. When a longitudinal division follows a transverse division, the plastid remains in its position adjacent to an old transverse wall where it is bisected by the plane of the longitudinally oriented preprophase band microtubules.     

Intramural ganglia of the human gallbladder: morphological and functional observations]

The purpose of this study was the reinvestigation of the intrinsic innervation of human gall bladder with an immunohistochemical technique named peroxidase anti-peroxidase (PAP). The antigen demonstrated was the S100 protein normally present in the surface of glial cells, Schwann cells and satellite cells in ganglia. The tissues used were taken from 20 human gall bladders, fixed after surgery. This technique is not specific to demonstrate adrenergic or cholinergic innervation but it reveals just myelinated fibers. The current study was undertaken in order to study the organization and the function of plexus of nerves and ganglia present in the wall of the gall bladder. The neck of the gall bladder was the region in which the higher number of nerve cells and nervous fibers was present. The technique used has demonstrated ganglionated plexus and nerves in submucosal layer, fibromuscular and adventitial layer according to the enteric nervous system. All ganglia are postganglionic stations related with preganglionic cholinergic fibers. These results confirm that the intramural ganglia of the gall bladder are analogous to those of the enteric nervous system according to their common origin.     

Distribution and anatomy of GABA-like immunoreactive neurons in the central and peripheral nervous system of the snail Helix pomatia

Gamma-aminobutyric acid (GABA)-like immunoreactive neurons were studied in the central and peripheral nervous system of Helix pomatia by applying immunocytochemistry on whole-mount preparations and serial paraffin sections. GABA-immunoreactive cell bodies were found in the buccal, cerebral and pedal ganglia, but only GABA-immunoreactive fibers were found in the viscero-parietal-pleural ganglion complex. The majority of GABA-immunoreactive cell bodies were located in the pedal ganglia but a few could be found in the buccal ganglia. Varicose GABA-ir fibers could be seen in the neuropil areas and in distinct areas of the cell body layer of the ganglia. The majority of GABA-ir axonal processes run into the connectives and commissures of the ganglia, indicating an important central integrative role of GABA-immunoreactive neurons. GABA may also have a peripheral role, since GABA-immunoreactive fibers could be demonstrated in peripheral nerves and the lips. Glutamate injection did not change the number or distribution of GABA-immunoreactive neurons, but induced GABA immunoreactivity in elements of the connective tissue ensheathing the muscle cells and fibers of the buccal musculature. This shows that GABA may be present in different non-neural tissues as a product of general metabolic pathways.     

Optimized transection: a prelude to oriented sectioning

A technique is described which permits blocks of tissue to be flat-embedded in euhedral plastic castings and then to be transected along a plane so that sections may be cut which are optimally oriented to the internal ultrastructure of the block. In the transection procedure a hollow plastic cylinder is placed on the specimen trimming block. The cylinder's top prescribes a plane to which the tissue block is accurately oriented and clamped at a predetermined level. Two hand files and a burnisher are worked across the cylinder's top to 1) remove extraneous material above the plane of transection, 2) expose the tissue for sectioning and 3) smooth the block face. The clear plastic at the periphery of the exposed tissue is then easily trimmed away with a razor blade. The result is a block face with a flat, reflective surface which may be quickly aligned to the knife on the ultramicrotome. The effort needed to transect, align and face the block is minimal and 1-micron or semithin sections produced will be precisely parallel to, and at, the plane of transection. Dust produced by the transection procedure is easily eliminated from the work area by use of a small disposable vacuum cleaner. The technique of producing optimally oriented light microscope sections, using the transector, is enhanced by application of solvents to the block face which cause it to develop a temporary low relief, exactly matching the structural detail of sections cut from the block face.(ABSTRACT TRUNCATED AT 250 WORDS)     

The conformation and orientation of copper(II)-bleomycin intercalated with DNA

EPR data are used to describe the conformation and identity of the atoms coordinated to Cu(II) in Cu(II)-bleomycin bound to oriented DNA fibers. The fibers were slowly drawn from viscous solutions of Cu(II)-bleomycin-DNA containing one Cu(II)-bleomycin to 200 basepairs. EPR measurements were made at room temperature and 90 K for different orientations of the external magnetic field with respect to the helical axes of the fibers. The g-values ($\text{g}{}_{\mathrm{}}\text{=2.21}$, g_⊥ =2.04) and the hyperfine constant ($\text{A}{}_{\mathrm{}}\text{=175}\text{G}$) are consistent with values expected for Cu(II) chelated to a square planar array of ligands. In the oriented fibers, the square planar arrays do not all have the same orientations with respect to the fiber axes. At room temperature the chelated ions have rotational freedom in which the normal to the planar array has almost complete freedom of rotation about axes perpendicular to the DNA fiber axes. The normal maintains an angle of 75° with respect to the axis, in the plane of the basepair, about which it rotates. Nine superhyperfine peaks on the high field side of the EPR spectrum were partially resolved. The number and splitting (12 G) of these superhyperfine peaks indicate that four nitrogen atoms are chelated to Cu(II) in a square planar array. These data on Cu(II)-bleomycin bound to DNA give information on the orientation of the metal-containing portion of bleomycin which lies outside the double helix.     

异丙肾上腺素对心内神经节中肽能递质VIP影响

异丙肾上腺素是临床常用的心脏骤停的抢救药物。为了研究该药对心内神经节中肽能递质的影响,本文在大鼠皮下注射异丙肾上腺素５ｍｇ／ｋｇ,连续三天,后固定取心房后壁,用免疫组化结合图像分析,观察心内神经节中肽能递质ＶＩＰ的变化。对照组大鼠心内神经节中含有ＶＩＰ免疫反应（ＶＩＰ－ＩＲ）阳性神经纤维和胞体;实验组大鼠心内神经节中含有ＶＩＰ－ＩＲ阳性神经纤维和胞体呈不同程度增多。其中ＶＩＰ－ＩＲ阳性神经纤维积分光密度较对照组增加２５．３％,而神经胞体积分光密度只增加８．１％。结果提示：１．大鼠心内神经节中ＶＩＰ－ＩＲ阳性神经纤维可能有两个来源：即心内ＶＩＰ－ＩＲ阳性神经节细胞和心外副交感神经元;２．异丙肾上腺素对心脏的作用并非单一的直接作用,其中部分是通过影响心内神经节中肽能递质的变化而发挥间接作用。     

Pathways for the transmission of visual information in the protocerebrum of the drone fly Eristalis tenax]

Studies have been made on the structure of neuropiles and visual pathoways in the brain of the fly E. tenax L. (Diptera, Syrphidae). The retina is projected on laminar structures in the visual ganglia only; other protocerebrum neuropiles lack this projection. All the comissures connecting contralateral visual ganglia, consist of several hundreds of fibers, whereas the binocular zone of both eyes includes more than 4,000 ommatidia. Neither the visual ganglia, nor other protocerebrum neuropiles may serve as a substrate for topographic imposition of projections of the corresponding parts in both retines. The mechanism of binocular interaction in insects presumably differs from that in mammals (primates, carnovores).