Structure of the intramural nerves of the rat bladder

The bladder of adult female rats receives ～16,000 axons (i.e., is the target of that many ganglion neurons) of which at least half are sensory. In nerves containing between 40 and 1200 axons cross-sectional area is proportional to number of axons; >99% of axons are unmyelinated. A capsule forms a seal around nerves and ends abruptly where nerves, after branching, contain ～10 axons. A single blood vessel is present in many of the large nerves but never in nerves of <600 axons. The number of glial cells was estimated through the number of their nuclei. There is a glial nucleus profile every 76 axonal profiles. Each glial cell is associated with many axons and collectively covers ～1,000 μm of axonal length. In all nerves a few axonal profiles contain large clusters of vesicles independent of microtubules. The axons do not branch; they alter their relative position along the nerve; they vary in size along their length; none has a circular profile. All the axons are fully wrapped by glial cells and never contact each other. The volume of axons is larger than that of glial cells (55%–45%), while the surface of glial cell is twice as extensive as that of axons; there are ～2.27 m² of axolemma and ～4.60 m² of glial cell membrane per gram of nerve. Of the mitochondria of a nerve ～3/4 are in axons and ～1/4 in glial cells.     

Freeze-fracture observations on the plasma membrane,the cell wall and the cuticle of growing protonemata of Adiantum capillus-veneris L.

Using freeze-fracture electron microscopy we have examined the morphology of the plasma membrane and the cell wall of single-celled protonemal filaments of the fern Adiantum capillus-veneris grown under continuous red light. The surface of the protonemal cell wall is completely covered by a multilayered, lipid-like coat, probably consisting of cuticular waxes. The rhizoid seems to lack this type of coat. The cell walls of the protonemata contain 8-nm thick, randomly oriented fibrils. In rapidly growing protonemata the P-face of the plasma membrane contains both randomly distributed particles and distinct particle rosettes. The rosettes consist of six 8–9-nm-wide particles in a ring-like configaration and have an outer diameter of 24 nm. They closely resemble the particle rosettes seen on the P-face of the plasma membrane of green algae and of higher plants, which recently have been implicated in the synthesis of cellulose fibrils. Within 20 m from the tip of the protonemata, and coinciding with the region of maximal cell-wall growth and expansion and thus cellulose-fibril synthesis, the greatest density of rosettes (20/m²) is observed. Beyond 20 m from the tip this number drops rapidly to near zero at 50 m. The rosettes have a tendency to form small, irregular clusters, but only very rarely are three or more rosettes found in a row or in a geometrical pattern. Our measurements of the size and the density of the randomly distributed plasma membrane particles indicate that the tip region must be specialized with respect to other plasma-membrane activities as well. Thus the tip region contains not only the highest density of randomly destributed intramembrane particles, but also particles of different sizes than those found elsewhere in the plasma membrane.     

Astrocytes alter their polarity in organotypic slice cultures of rat visual cortex

The ultrastructure of astrocytes in an organotypic slice culture of the rat visual cortex was investigated using ultrathin sections and freeze-fracture replicas. After a culture period of 9–15 days, a glial scaffold formed that separated the bulk of the slice neuropil from the medium and the underlying plasma clot. However, the glial cells and processes did not build a dense barrier but allowed the outgrowth of neurites. A basal lamina covering the medium-oriented surface of the astrocytes was not found. In freeze-fracture replicas, orthogonal arrays of particles (OAP) were characteristic components of astrocytic membranes. The OAP density in membranes bordering the medium was 35±13 OAP/m², corresponding to 2.5% of this membrane area; the OAP density in membranes within the slice neuropil was 22±12 OAP/², corresponding to 1.4% of this membrane area. Although the difference was significant, it was greatly reduced when comparing OAP densities in endfoot and non-endfoot membranes in vivo. Another mode of polarity was recognized in astrocytes of the organotypic slice culture. In membranes of astrocytes bordering upon the medium, the density of non-OAP intramembranous particles (IMP) was clearly higher (1130±136 IMP/ m²) than in membranes of astrocytes in the center of the slice (700±172 IMP/m²). This pronounced IMP-related polarity was observed neither in vivo nor in cultured astrocytes. The present study suggests, together with data from the literature, that the distribution of astrocytic OAP across the cell surface is influenced by the existence of a basal lamina and neuronal activity, and that astrocytes possess a more remarkable plasticity of membrane structure than previously suspected.     

An electron microscopical study of the ventral nerve cord of the leech

Summary Three types of fibrillar structure can be seen with the electron microscope in nerve cells of the vental nerve cord of the leech: the neurofibrillar bundles, the tubules and the tonofibrils. In neuroglial cells only the tonofibrils are present. The three types are structurally distinct, and, contrary to past suggestions, there is no evidence that neurofibrillar bundles may consist of tightly packed or badly fixed tubules.In vertebrates the electron microscope reveals bundles of discrete neurofilaments that form the basis for the argyrophilic neurofibrillae seen by light microscopy. Each neurofilamentous unit appears as a dot in cross section. In contrast, in the leech, the electron microscope shows compact fibrillar bundles that clearly correspond to the neurofibrils described by light microscopists. These bundles are made up of closely packed units rather than discrete filaments and where the units occur singly they are seen to have an angular or stellate outline in cross section. To make this distinction clear these have been termed neurofibrillar bundles rather than neurofilaments.Attachment plaques occur in both neurons and neuroglia. These plaques have tonofibrils attached, and the glial tonofibrils are far more numerous than the neuronal tonofibrils. The glial fibrils are identical with the tonofibrils in the glial cells.The attachment plaques are invariably related to an extracellular space that contains material identical with the basement membrane. This material is continuous, by a complex system of channels and diverticulae, with the outer basement membrane in the neuron packets, but forms isolated patches in the other parts of the nervous system.We are grateful to Prof. J. Z. Young, F. R. S., for his encouragement to Mrs. Astafiev for the drawings, to Miss B. Shirra and Mr. K. Watkins for technical assistance and to Mr. S. Waterman for photography.     

Qualitative and quantitative comparison of the distribution of phosphorylated and non-phosphorylated neurofilament epitopes within central and peripheral axons of adult hamster (Mesocricetus auratus)

Summary The distribution of phosphorylated and nonphosphorylated neurofilament epitopes was determined immunocytochemically in adjacent 2 m-thick sections of sciatic nerve, ventral root and spinal cord. Staining was scored as either intense, moderate or absent and the proportion of labeled axons was calculated for each category. Nearly all sciatic nerve and ventral root axons were immunoreactive with both antibodies against phosphorylated and non-phosphorylated neurofilaments and there were no significant differences in the number of intensely- or moderately-labeled axons. Within the spinal cord however, while the majority of large caliber axons was stained with both antibodies, there was a significant number of small caliber axons which stained only with antibodies against phosphorylated neurofilaments. These results show that phosphorylated and nonphosphorylated neurofilaments are extensively codistributed in CNS and PNS axons, and that in the CNS, staining intensity for non-phosphorylated epitopes is less in the smaller axons.     

ELECTRON MICROSCOPE AND EXPERIMENTAL INVESTIGATIONS OF THE NEUROFILAMENTOUS NETWORK IN DEITERS'' NEURONS : Relationship with the Cell Surface and Nuclear Pores

The assembly of filamentous elements and their relations to the plasma membrane and to the nuclear pores have been studied in Deiters' neurons of rabbit brain. Electron microscopy of thin sections and of ectoplasm spread preparations have been integrated with physicochemical experiments and differential interference microscopy of freshly isolated cells. A neurofilamentous network extends as a continuous, three-dimensional, semilattice structure throughout the ectoplasm, the "plasma roads," and the perinuclear zone of the perikaryon. This space network consists of ~90-Å wide neurofilaments arranged in fascicles which are interconnected by an exchange of neurofilaments. The neurofilaments consist of intercoiled ~20-Å wide unit-filaments and are associated through cross-associating filaments with other neurofilaments of the fascicle and with microfilaments. The ~20–50-Å wide microfilaments display intimate associations with the plasma membrane and with the nuclear pores. Electron microscopy of thin sections from glycerinated and heavy meromyosin-treated Deiters' neurons shows that actin-like filaments are present in the pre- and postsynaptic regions of synapses terminating on these neurons. It is proposed that the neurofilamentous space network serves a transducing function by linking plasma membrane activities with the genetic machinery of the neuron.     

Intracellular chloride activity of leech neurones and glial cells in physiological,low chloride saline

Leech blood apparently contains considerably less chloride than generally used in physiological experi ments. Instead of 85–130 mM Cl^– used in experimental salines, leech blood contains around 40 mM Cl^– and up to 45 mM organic anions, in particular malate. We have reinvestigated the distribution of Cl^– across the cell membrane of identified glial cells and neurones in the central nervous system of the leech Hirudo medicinalis L., using double-barrelled Cl^–- and pH-selective micro electrodes, in a conventional leech saline, and in a saline with a low Cl^– concentration (40 mM), containing 40 mM malate. The interference of anions other than Cl^–to the response of the ion-selective microelectrodes was estimated in Cl^–-free salines (Cl^– replaced by malate and/or gluconate). The results show that the absolute intracellu lar Cl^– activities (aCl_i) in glial cells and neurones, but not the electrochemical gradients of Cl^– across the glial and the neuronal cell membranes, are altered in the low Cl^–, malate-based saline. In Retzius neurones, aCl_i is lower than expected from electrochemical equilibrium, while in pressure neurones and in neuropil glial cells, aCl_i is distributed close to its equilibrium in both salines, re spectively. The steady-state intracellular pH values in the glial cells and Retzius neurones are little affected (0.1 pH units) in the low Cl^–, malate-based saline.     

Cytochemical demonstration of nucleoside phosphatase activity in myelinated nerve fibers of the rat

Summary The histochemical and cytochemical localization of end product from the Wachstein-Meisel reaction was examined in transverse frozen sections of rat ischiatic nerve and spinal nerve roots. A variety of fixatives, substrates, and inhibitors were used at varying pH. Following glutaraldehyde fixation, nucleoside phosphatase activity was noted in the axon-Schwann cell interface of both unmyelinated and myelinated nerve fibers. The presence of this enzymatic activity at this strategic location in the nerve fiber complex attests to the importance of this space as a metabolically active site.Additional deposits of end product occurred on neurofilaments of myelinated and unmyelinated axons and were observed as diffuse axonal stains by light microscopy. These precipitates had a predilection for tissue fixed in formalin or hydroxyadipaldehyde and were relatively more prominent following acidic incubations. The resemblance between these deposits and spurious axonal acid phosphatase precipitates was discussed.This study was supported by Grant NB 04161 of the National Institute of Neurological Disease and Blindness.     

“Tight” junctions in the sheath of normal and regenerating motor nerves of the crayfish,Orconectes virilis

Summary Tight or occluding intercellular junctions occur between adjacent glial processes in normal and regenerating crayfish motor nerve sheaths. Although infrequent, these junctions possess the ridge and groove configuration characteristic of freeze-cleaved occluding junctions. When present, nerve sheath tight junctions consist of a single, or at most a few, parallel intramembrane ridges situated on the EF membrane face of the glial plasma membrane. Consequently, such contacts are rarely recognized in thin sections of plasticembedded nerve sheaths. Crayfish nerve sheath tight junctions are of the fascia occludens type and, therefore, do not impede solute flow across the nerve sheath. Fasciae occludentes of regenerating nerve sheaths occur in close proximity to discoid plaque-like aggregates of particles assumed to represent maculae adhaerentes. This relationship, which was not observed in normal nerve sheaths, suggests a functional association between the two types of junctions, perhaps developmental transformation of one junction type into the other. Although ridges and grooves of tight junctions occur next to crossfractured trans-glial channels, no functional significance is proposed for this relationship. This study is the first report of tight intercellular junctions in crustacean glial nerve sheaths.Supported by the National Research Council of Canada     

Beneficial effects and mechanism of action of Momordica charantia juice in the treatment of streptozotocin-induced diabetes mellitus in rat

This study investigated the beneficial effects and mechanism of action of the juice of Momordica charantia in streptozotocin (STZ)-induced diabetes mellitus in rats. Diabetes mellitus was associated with significant (p < 0.01) time course reductions in body weight, plasma insulin and the number of insulin positive cells per islet and significant (p < 0.01) time course elevation in blood glucose and osmolarity and systolic blood pressure compared to age-matched healthy controls. Oral intake of M. charantia juice by STZ-induced diabetic rats partially reversed all the diabetes-induced effects measured. Daily oral administration of M. charantia juice to STZ-induced diabetic rates significantly (p < 0.01) reduced the Na⁺- and K⁺ -dependent absorptions of glucose by the brush border membrane vesicles of the jejunum compared to the responses obtained in STZ-induced diabetic rat. Either insulin (100 MM) or the fruit juice lyophilised extract (5 g · ml^–1) can stimulate ¹⁴C-D-glucose uptake in L6 myotubes. These effects were completely blocked by wortmannin, an inhibitor of phosphatidylinositol 3-kinase. High concentrations (10–200 g · ml^-1) of M. charantia juice extract inhibited ¹⁴C-D-glucose uptake in L6 myotubes compared to the control response. The effect of M. charantia treatment was also investigated on myelinated fibre abnormalities in the tibial nerve of STZ-induced diabetic and control rats. The results show that diabetes was associated with significant (p < 0.05) reduction in the mean cross-sectional myelinated nerve fibres, axonal area, myelin area and maximal fibre area compared to end controls. Treatment of STZ-induced diabetic rats with M. charantia juice normalised the structural abnormalities of peripheral nerves. The results indicate that M. charantia can exert marked beneficial effects in diabetic rats, and moreover, it can regulate glucose uptake into jejunum membrane brush border vesicles and stimulate glucose uptake into skeletal muscle cells similar to the response obtained with insulin. (Mol Cell Biochem 261: 63–70, 2004)     

Density of Thy-1 on Axonal Membrane of Different Rat Nerves

Abstract: The density of the cell surface antigen, Thy-1, has been measured on the axonal plasma membrane of the optic and hypoglossal nerves, and on the sympathetic (preganglionic) chain of the superior cervical ganglion in the rat. For each, the amount of Thy-1 in a standard length of nerve was determined. The amount of axonal plasma membrane in a similar length was then measured by use of a computer graphics system which determined the total length of membrane in electron micrographs of cross sections of nerve. The axons of the two peripheral nerves were found to have a similar density (1100–1500 molecules/μm²) of Thy-1 on their surface, and this was two- to threefold higher than the density on axons of optic nerve (500 molecules/μm²). These figures indicate that the density of Thy-1 on the surface of these axons is somewhat lower than that found on the cells of the lymphoid system. Moreover, contrary to the impression gained from previous determinations of Thy-1 levels, axons in the peripheral nervous system do not necessarily have a lower density of Thy-1 on their surface than do those of the central nervous system.     

Specific binding of the calcium channel blocker [3H]verapamil to membrane fractions of Chlamydomonas reinhardtii

Specific binding of the calcium antagonist [³H]verapamil to a microsomal fraction, a presumptive plasma membrane fraction and an intracellular membrane fraction of the phototactic unicellular green alga Chlamydomonas reinhardtii has been demonstrated. The specific activity of the plasma membrane marker enzyme K⁺-stimulated, Mg²⁺-dependent ATPase was severalfold higher in the upper (polyethylene glycol-rich) than in the lower (dextran-rich) phase, and the reverse was established for the marker enzymes of intracellular membranes such as cytochrome c oxidase for mitochondria and antimycin Aresistant NADPH-cytochrome c reductase for endoplasmic reticulum. Chlorophyll as a marker for thylakoid fragments was exclusively found in the lower phase. In the microsomal fraction two specific binding sites of [³H]verapamil were found at 22°C, one with higher and a second with lower affinity to [³H]verapamil. Separation of plasma membranes from intracellular membranes revealed that the highaffinity binding site is attributed to the plasma membrane fraction whereas the low-affinity binding site can be attributed to the intracellular membrane fraction. Specific binding to both separated membrane fractions is saturable and reversible. [³H]Verapamil binding to plasma membranes was not inhibited by the calcium channel blockers diltiazem and nifedipine. However, in the intracellular membrane fraction [³H]verapamil could be displaced by diltiazem but not by nifedipine. Increasing concentrations of calcium chloride inhibited [³H]verapamil binding in both fractions.Abbreviations B_max maximum density of binding sites - BSA bovine serum albumin - Cyt.c cytochrome c - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - EGTA ethyleneglycol-bis(2-amino-ethylether)N,N-tetraacetic acid - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - IC₅₀ concentration causing 50% inhibition - Mes [N-morpholino]ethanesulfonic acid - PEG polyethylene glycol - PMSF phenylmethylsulfonylfluoride - PVPP polyvinylpolypyrrolidone - TCA trichloroacetic acid     

Occurrence of innervation in labral glands of Daphnia obtusa (crustacea,cladocera)

Electron microscopy discloses nerve endings in contact with gland cells situated in the labrum of Daphnia. Swellings of nerve fibers are in close contact with gland cell membranes, either on the cell surface or inserted into infoldings of plasma membrane. The axonal processes are single or double and lack glial wrappings. Inside the nerve fibers are vesicles of different sizes and electron density. These include granular vesicles, which often are dense-cored, and also clearer vesicles. Some presynaptic differentiations lie along the contact line of the axonal process with the gland cell membrane. The significance of the vesicles is discussed in terms of their possible content of biogenic amines, as described in other invertebrates.     

Synaptic organisation of the pelvic ganglion in the guinea-pig

Summary A semi-quantitative electron-microscopic study of neuronal cell bodies, nerve profiles and synapses in the anterior pelvic ganglia of the guinea-pig has been carried out following in vivo labelling of adrenergic nerve endings with 5-hydroxydopamine. Ganglion cells of three main types have been distinguished: 1) the majority (about 70%) not containing granular vesicles, probably cholinergic elements; 2) those containing large granular vesicles of uniform size (80–110 nm), with granules of medium density and prominent halos; and 3) those containing vesicles of variable size (60–150 nm), with very dense eccentrically placed granular cores. Some non-neuronal granule-containing cells were present, mainly near small blood vessels. Some 95% of the total axon profiles within the ganglia were cholinergic, the remaining 5% were adrenergic. However, 99% of synapses (i.e. axons within 50 nm of nerve cell membrane with pre-and post-synaptic specialisations) were cholinergic, and 1 % were adrenergic. Only three examples of nerve cell bodies exhibiting both cholinergic and adrenergic synapses were observed. Unlike the para-and prevertebral ganglia, the pelvic ganglia contained large numbers of axo-somatic synapses. As many as 20% of the nucleated neuronal cell profiles displayed two distinct nuclei.     

Structure and ultrastructure of the frog motor endplate

Summary The frog motor endplate in its simplest form consists of an elongated, slender nerve ending embedded in a gutter-like depression of the sarcolemma. This nerve terminal contains the usual synaptic organelles. It is covered by a thin coating of Schwann cell cytoplasm which embraces the terminal with thin finger-like processes from both sides, thereby sub-dividing it into 300–1000 regularly spaced compartments. The individual synaptic compartments correspond to the strings of varicosities or grape-like configurations of motor nerve terminals in endplates of other species and in the cerebral neuropil of vertebrates.Each compartment contains one or more bar-like densities of the presynaptic membrane, active zones, which are associated with the attachment sites between synaptic vesicles and plasmalemma. Active zones have a regular transverse arrangement and occur at specific loci opposite the junctional folds. The attachment sites for synaptic vesicles are at the edges of the bars which are bilaterally delineated by a double row of 10 nm particles attached to the A-face. The structural appearance of vesicle attachment sites in freeze-etch replicas corresponds to that of micropinocytosis. The active zones are often fragmented and the frequency of their association with vesicle attachment sites is highly variable.The junctional folds are characterized by specific sites in which intramembranous particle aggregations occur at relatively high packing density (7500/m²). These sites are located opposite the active zones at the juxtaneural lips, a location where one would expect ACh-sensitive receptors on the postsynaptic membrane.This work was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 38, Projekt N), The Swiss National Foundation for Scientific Research (Grants Nr. 3 82372 and 3 77472) and the Dr. Eric Slack-Gyr Foundation Zürich.     

再生神经中微管,神经丝与轴突截面积的变化

用电镜及图象分析的方法研究了再生轴突中微管、神经丝与轴突截面积的变化,发现神经再生过程中微管及神经丝的密度增加,并与轴突截面积呈相关关系,而且微管的变化更早,更明显。由于微管参与了轴浆转运的机制,微管的增加提示其在神经再生中起了重要的作用。     

The cytoskeleton of Cobaea seed hairs:

The cell wall of Cobaea scandens seed hairs developed in a characteristic sequence, with the deposition of a cellulose thread onto a pectic swelling layer was the final event. The cellulose thread was intracellularly accompanied by a band of 10–18 microtubules. During the formation of the swelling layer the microtubules were homogeneously distributed; they ran circumferentially normal to the cell axis. When cellulose-thread formation started, the microtubules became arranged in a helical band. The density of the microtubules varied during the different phases of development. The highest density was observed before cellulosethread formation and ranged from 6–15 m·m^-2. The length of the microtubules, 20–30 m, was determined by direct measurements, as well as estimated from the total microtubular length in a given area and the counted free ends. With the indirect immunofluorescence technique the microtubules of the band stained inhomogeneously. Those which were located at the edges of the band fluoresced more intensely than those of the central part. Attempts to visualize actin filaments in the hair cells with rhodaminyl-conjugated phalloidin resulted in a homogeneous staining of the area of the microtubular band, indicating that actin filaments may be present in this region. Though, in thin sections and dry-cleaved cells, filamentous structures were observed between the microtubules, caution is expressed that the observed fluorescence was, indeed, due to actin filaments. The role of the filamentous structures is discussed with respect to formation and maintenance of the microtubular band. Microtubules apparently did not cross coated pits which were visualized in the plasma membrane through the dry-cleaving technique.Abbreviations IFT indirect immunofluorescence technique - RP rhodaminyl-conjugated phalloidin - SEM scanning electron microscopy     

Protein turnover in the attached leaves of non-stressed and stressed barley seedlings

Protein turnover was examined, using tritiated water, in various 2-cm regions of 7-11-d-old, first leaves of barley (Hordeum vulgare). Differences were found between the regions in their protein turnover and their responses to stress. The rate constant for degradation for total protein was the same throughout the leaf and the average half-life (t_1/2) of protein=approx. 220 h. Only in the older regions did a 24-h pulse of³H₂O preferentially label protein with a t_1/2 (90 h) considerably shorter than the t_1/2 for total protein. Soluble protein was degraded faster than insoluble protein and contained an appreciable short-lived protein component observable by short-pulse labelling. The rate of protein synthesis was greatest in the cells of the youngest region and declined as each region aged. The mean rate of protein synthesis over the 4-d period was 4 and 7 nmol h^-1 of amino-N with respect to the regions 1–3 and 7–9 cm from the leaf tip. Seedlings, stressed by adding polyethylene glycol (2.0 MPa) to the roots, showed a marked loss of protein from the older leaf regions with only small losses in the younger regions. Amino acids accumulated in the younger region continuously whereas in the older region little accumulation occurred until day 3 of stress when proline levels increased. Protein synthesis was decreased by between 30% and 50% in all leaf regions. In the region 1–3 cm from the leaf tip, the rate of protein degradation of total protein was enhanced and equalled the rate of degradation of 24-h-pulse-labelled protein which was not itself significantly affected by stress (t_1/2=approx. 90 h). In the region 3–5 cm, the degradation of both 4-d and 24-h-labelled protein was enhanced by stress to rates similar to those found in the region 1–3 cm. This was largely through increases in the degradation of the insoluble protein, but the degradation of soluble protein was also raised. Protein degradation in the region 7–9 cm was not affected by stress.Abbreviations t_1/2 average half-life - PEG polyethylene glycol     

DISTRIBUTION OF LEUCINE-3H DURING AXOPLASMIC TRANSPORT WITHIN REGENERATING NEURONS AS DETERMINED BY ELECTRON-MICROSCOPE RADIOAUTOGRAPHY

The distribution of leucine-³H in neurons was determined by electron-microscope radioautography after infusion of label into the spinal cord or sensory ganglia of regenerating newts. In the nerve cell bodies 3 days after infusion, the highest concentration of label per unit area occurred over the rough-surfaced endoplasmic reticulum. In the large brachial nerves, the silver grains were not distributed uniformly in the axoplasm, indicating that the labeled materials are restricted in their movement to certain regions of the axon. Almost all of the radioautographic grains observed in myelinated nerves could be accounted for by the presence of a uniformly labeled band occupying the area 1500–9000 A inside the axolemma. This region of the axon was rich in microtubules and organelles while the unlabeled central core of the axon contained mainly neurofilaments. This observation supports the hypothesis that microtubules are related to axonal transport. In small, vesicle-filled nerve terminals in the blastema, labeled material was restricted to a thin zone a short distance beneath the plasma membrane while the central region of the terminal was largely unlabeled. The peripheral pattern of labeling in the nerve endings is consistent with successive addition of newly synthesized proteins at the periphery of the growth cone and release of substances such as trophic factors at the nerve terminal.     

Capacitance,short-circuit current and osmotic water flow across different regions of the isolated toad skin

Summary The amphibian antidiuretic hormone, arginine vasotocin, stimulated osmotic water flow across isolated skin from the pelvic but not the pectoral skin of the toad, Bufo woodhouseii. Changes in the apical membrane capacitance were not observed for either region of the skin following treatment with arginine vasotocin when there was an osmotic gradient across the tissue. In the absence of an osmotic pressure gradient, the apical membrane capacitance of the pelvic skin increased from 2.8±0.5 to 3.3±0.6 F · cm^-2 after treatment with 5 · 10^-8 M arginine vasotocin. Under these conditions, apical membrane capacitance of the pectoral skin was 1.8±0.1 F · cm^-2 and did not change significantly after arginine vasotocin treatment. The amiloride-sensitive short-circuit current across the pelvic skin was stimulated by arginine vasotocin as was the density of channels in the apical membrane as determined by fluctuation analysis. Values for channel density in the pelvic skin also correlated with apical membrane capacitance and increased from 90 to 273 channels per m² of estimated membrane area following arginine vasotocin treatment. In the pectoral skin the stimulation of short-circuit current following arginine vasotocin treatment was small and an increase in channel density could not be demonstrated. The current through single Na⁺ channels in both regions of the skin did not different either before or after arginine vasotocin treatment.Abbreviations A amiloride - ADH antidiuretic hormone - AVT arginine vasotocin - C capacitance - C _a capacitance of apical membrane - f _c corner frequency - i single-channel current - osmotic water flow - IMP intramembrane particles - I _sc short-circuit current - amiloride-sensitive short-circuit current - M channel density - P _o probability of a channel being open - R channel receptor - R _a apical resistance - R _p paracellular resistance