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Biomedical subjects

D Purves

Publications and source records attributed to D Purves.

106 records · Page 6Linked to original sources
PubMed1976

Visual identification of two kinds of nerve cells and their synaptic contacts in a living autonomic ganglion of the mudpuppy (Necturus maculosus).

1. Many of the nerve cells comprising the cardiac parasympathetic ganglion of the mudpuppy are spread out in a thin, transparent sheet of tissue, enabling one to see cellular details in living preparations with differential interference contrast optics. The aim of this study was twofold: to establish the morphology of the nerve cells and their synaptic connections by light and electron microscopy, and to determine which aspects of the ganglion's structure could be reliably identified in the living tissue. 2. There are two types of neurones in the ganglion: (a) principal cells that send post-ganglionic axons to cardiac muscle fibres, and (b) interneurones whose processes are confined to the ganglion. 3. Interneurones are distinguished from principal cells by the presence of numerous granular vesicles seen with the electron microscope, and by intense formaldehyde-induced fluorescence. The interneurones are thus similar to catecholamine-containing interneurones in autonomic ganglia of other vertebrates. 4. Principal cells are innervated by processes that terminate mainly on the cell body, forming up to forty-five synaptic boutons and covering, on the average, 5% of the perikaryal surface. The synaptic terminals are derived from three sources: (a) axons from the vagus nerves, (b) interneurones and (c) other principal cells. Vagal terminals contacting principal cells contain agranular vesicles typical of preganglionic cholinergic endings. At regions of contact between processes of interneurones and principal cells, the interneurones have granular vesicles focused at membrane specializations; in addition there are small areas of close plasma membrane apposition, probably gap junctions. Some of the contacts between principal cells are characterized by gap junctions; others are structurally similar to vagal endings but persist after vagal degeneration. 6. Interneurones are innervated by axons that make contact mainly with their processes. The axon terminals on processes of interneurones contain agranular vesicles similar to vagal terminals on principal cells. 7. In live preparations principal cells are distinguished from interneurones by their size and the appearance of their organelles. Synaptic contacts on principal cells could often be identified and, in some cases, large contacts from interneurones or those from other nearby principal cells could be traced back to their cell bodies of origin. The validity of these identifications was confirmed by subsequent electron microscopic examination of the same cells.

Animals↗
PubMed1976

Functional and structural changes in mammalian sympathetic neurones following colchicine application to post-ganglionic nerves.

1. The effects of post-ganglionic colchicine application on neurones of the guinea-pig superior cervical ganglion were studied with intracellular recording and electron microscopy. 2. Local colchicine application for 30 min to one of the major post-ganglionic nerves caused several electrophysiological changes after 4-7 days in many neurones whose axons run in this nerve. These changes include: (a) a reduction in the amplitude of synaptic potentials elicited by supramaximal preganglionic stimulation; (b) a decrease in the number of preganglionic fibres innervating individual neurones; (c) the development of regenerative responses in dendrites; and (d) the failure of antidromic action potentials to fully invade the neuronal soma. These functional changes occurred in the absence of impaired impulse conduction or axon degeneration, and were not observed in nearby neurones whose axons ran in an untreated post-ganglionic nerve. The effects of colchicine are similar to the changes produced by axotomy. 3. Counts of synapses in thin sections from the region of the ganglion where the affected neurones were located showed a reduction, compared to the number of synapses in other regions of the colchicine treated ganglia, or normal control ganglia. This finding indicates that synaptic depression after colchicine treatment, like that after axotomy, is due primarily to a loss of synaptic contacts from the dendrites of affected nerve cells. Unusual profiles containing numerous vesicular and tubular organelles frequently seen after interruption of the axons were also observed in thin sections after colchicine treatment. 4. The similarity of the electrophysiological and ultrastructural effects of colchicine treatment and axon interruption offers further support for the view that synaptic contacts on sympathetic neurones are normally regulated by an interaction of the neuronal soma with its axonal extension to the periphery.

Action Potentials↗
PubMed1976

Competitive and non-competitive re-innervation of mammalian sympathetic neurones by native and foreign fibres.

The ability of native (sympathetic preganglionic) and foreign (vagal) nerve fibres to re-innervate neurones of the guinea-pig superior cervical ganglion, either alone or in competition with each other, has been studied by means of intracellular recording and electron microscopy. 1. Native fibres make synaptic contacts with nearly all ganglion cells within one month of cervical trunk section; within 6 months the degree of innervation, judged by measurement of excitatory post-synaptic potential (e.p.s.p.) amplitude and electron microscopical synapse counts, approaches normal. However, even after 15 months innervation was weaker than in normal control ganglia. 2. Vagal fibres are less successful during re-innervation. Although a similar number of foreign fibres grown into denervated ganglia and make contact with nearly all ganglion cells within a month, after 6-12 months e.p.s.p. amplitudes in response to foreign nerve stimulation remain relatively small, and counts of synapses are only about 60% as great as in ganglia re-innervated with the native nerve. 3. When both native and foreign fibres are allowed to re-innervate ganglion cells simultaneously, about half the neurones in the ganglion receive synapses from both sources after 1 month. The proportion of dually invervated cells remains roughly constant for at least 14 months. Neither set of preganglionic fibres dominates or displaces the other, although neurones generally are re-innervated more effectively by native than foreign fibres, as is true during non-competitive re-innervation. 4. Thus during re-innervation of mammalian sympathetic neurones native fibres are preferred to foreign ones only in the sense that roughly the same number of native fibres form many more synapses on ganglion cells than do vagal axons. A foreign synapse, once formed, is as stable as a native one, and shows no tendency to be replaced by native terminals. These findings are discussed in relation to other evidence which has suggested specificity and selectivity during re-innervation of mammalian autonomic neurones.

Action Potentials↗
PubMed1975

Functional and structural changes in mammalian sympathetic neurones following interruption of their axons.

The effects of interrupting the axons of principal neurones in the superior cervical ganglion of adult guinea-pigs were studied by means of intracellular recording, and light and electron microscopy. 1. Within 72 hr of axon interruption, the amplitude of exitatory postsynaptic potentials potentials (e.p.s.p.s) recorded in principal neurons in response to maximal preganglionic stimulation declined. E.p.s.p.s were maximally reduced (by more than 70% on average) 4-7 days following interruption, and failed to bring many cells to threshold. E.p.s.p.s. recorded in nearby neurones whose axons remained intact were unaffected. 2. In ganglia in which axon interruption was achieved by means of nerve crush (thus allowing prompt regeneration), mean e.p.s.p. amplitudes began to increase again after about 1-2 weeks. One month after the initial injury many neurones had e.p.s.p.s of normal amplitude, and by 2 months affected neurones were indistinguishable from control cells. Functional peripheral connexions were re-established during the period of synaptic recovery. 3. The mean number of synapses identified electron microscopically in ganglia in which all the major efferent branches had been crushed decreased by 65-70% in parallel with synaptic depression measured by intracellular recording. However synapse counts did not return to normal levels even after 3 months. 4. During the period of maximum synaptic depression, numerous abnormal profiles which contained accumulations of vesicular and tubular organelles, vesicles, and mitochondria were observed in electron microscopic sections. Injection of horseradish peroxidase into affected neurones demonstrated dendritic swelling which probably correspond to these profiles. 5. Little or no difference was found in the electrical properties of normal neurones and neurones whose axons had been interrupted 4-7 days previously. However, the mean amplitude of spontaneously occurring synaptic potentials was reduced, and the amplitude distribution was shifted. This abnormality of the synapses which remain on affected neurones also contributes to synaptic depression. 6. Counts of neurones in normal and experimental ganglia showed that approximately half the principal cells died 1-5 weeks after crushing the major efferent brances. This finding presumably explains the failure of synapse counts to return to control levels after recovery. 7. If axons were prevented from growing back to their target organ by chronic ligation, surviving neurones whose axons were enclosed by the ligature did not generally recover normal synaptic function. Following ligation, most affected cells died within a month. 8. Thus the integrity of a principal cell's axon is necessary for the maintenance of preganglionic synaptic contacts, and ultimately for neuronal survival. The basis of neuronal recovery from the effects of axon interruption appears to be some aspect of regeneration to the peripheral target.

Animals↗
PubMed1974

The effect of contractile activity on fibrillation and extrajunctional acetylcholine-sensitivity in rat muscle maintained in organ culture.

1. The effect of contractile activity on the initiation of spontaneous action potentials (fibrillation) and on extrajunctional acetylcholine-sensitivity has been studied in single fibres in strips of previously denervated rat diaphragm maintained in organ culture for up to 10 days.2. Following removal of the diaphragm from the animal, fibrillation slowed and usually stopped altogether for about 24-36 hr. Thereafter, spontaneously active fibres were found in all cultured muscle strips.3. At any one time, about (1/4) to (1/3) of fibres impaled with micro-electrodes were active (defined as more than one action potential/10 sec), with a mean discharge frequency of 4.5/sec (range 0.1-24/sec).4. The duration of continuous activity in single fibres was, on average, 21-22 hr; a period of activity was followed by a longer inactive interval. Thus activity in single fibres is cyclical.5. Direct stimulation of fibrillating strips for 24 hr at 10/sec suppressed spontaneous activity for 1-3 days.6. Conversely, blockade of spontaneous activity with tetrodotoxin for 72 hr led to a two- to threefold increase in the number of fibrillating fibres when the drug was washed out; in some strips nearly all fibres became spontaneously active.7. The mean rate of activity of diaphragm fibres during normal breathing, determined by recording single units from the phrenic nerve in lightly anaesthetized animals, is about 18/sec.8. Direct stimulation of cultured diaphragm strips in a pattern similar to breathing for 7-8 days at an average rate of 10-12/sec (or 5/sec in some experiments), resulted in a marked reduction (about 95% in experiments at 10/sec) in extrajunctional sensitivity to ionophoretically applied ACh.9. Direct stimulation for 24 hr at 10/sec (comparable to a period of spontaneous activity) caused only a small reduction in extrajunctional ACh-sensitivity.10. We conclude that spontaneous activity in single fibres under these conditions occurs cyclically because activity, over a period of hours, inhibits the ability of the fibrillating fibre to initiate further action potentials. Repeated self-inhibition of spontaneous activity probably explains why denervated muscle fibres remain highly sensitive to extrajunctionally applied ACh.

Acetylcholine↗
PubMed1974

Membrane properties underlying spontaneous activity of denervated muscle fibres.

We have examined the events underlying the initiation of spontaneous action potentials (fibrillation) in fibres of previously denervated rat diaphragm maintained in organ culture for up to 10 days.1. Based on discharge pattern, two classes of spontaneously active fibres were found: rhythmically discharging fibres, and fibres in which action potentials occur at irregular intervals.2. Sites of action potentials initiation were located by exploration along the fibre length with two independent extracellular recording electrodes. The majority of sites of origin in both regular and irregular fibres were at the former end-plate zone; however, there was no region along the length that could not, at least in some fibres, be a site of origin.3. Intracellular recording at or near sites of origin of action potential discharge showed two types of initiating events. Irregularly discharging fibres were brought to threshold by discrete depolarizations of up to 15 mV in amplitude, while regularly occurring action potentials were associated with oscillations of the membrane potential.4. Discrete depolarizations (called fibrillatory origin potentials or f.o.p.s) at sites of origin in irregularly discharging fibres have the following properties: (a) random occurrence and nearly constant amplitude outside a refractory period during which both amplitude and probability of a second f.o.p. are reduced; (b) associated inward current flow which is localized to about 100 mum or less along the fibre length, and (c) dependence of amplitude and frequency on membrane potential.5. Oscillation of membrane potential found at sites of origin of action potential discharge in regular fibres also occurred locally along the fibre length and was sensitive to changes in membrane potential.6. Both f.o.p.s and oscillations of membrane potential were reversibly abolished by low Na(+)-Ringer fluid or tetrodotoxin.7. Neither type of initiating event was appreciably affected by concentrations of D-tubocurarine which blocked extrajunctional sensitivity to acetylcholine.8. We conclude that spontaneous action potentials under these conditions arise from a localized Na(+)-conductance change in the membrane of the active fibre; this conductance change is distinct from the increased Na(+)-conductance which follows the interaction of acetylcholine with its receptor. Spontaneous activity in single, denervated muscle fibres is cyclical and self-inhibiting (Purves & Sakmann, 1974); thus the Na(+)-conductance change underlying the initiation of spontaneous action potentials is affected by muscle fibre activity.

Action Potentials↗
PubMed1972

The distribution of synapses on a physiologically identified motor neuron in the central nervous system of the leech. An electron microscope study after the injection of the fluorescent dye procion yellow.

The fine structure of a physiologically identified motor neuron in the segmental ganglion of the leech central nervous system and the morphology of synapses on it were studied after injection of the fluorescent dye Procion yellow as a marker. The injected cell and its processes within the neuropil were located in thick or thin sections with fluorescence optics after initial fixation with glutaraldehyde and brief treatment with osmium tetroxide. The same or adjacent thin sections could then be examined in the electron microscope. Comparison with uninjected cells showed that the general features of the injected cell are retained although some organelles are distorted. The main features of the geometry of this neuron are the same from animal to animal: a single large process runs from the soma through the neuropil to bifurcate and enter the contralateral roots. Within the neuropil the main process gives off long branches (up to 150 micro), but these are greatly outnumbered by short branches and spines, one or a few microns in length, which were not appreciated in previous light microscope studies after injection of Procion yellow. Serial thin sections of selected areas along the main process within the neuropil showed that there are synapses on most of the shorter branches and spines; occasional synaptic contacts were also made on the main process itself and on longer branches. At least two morphologically distinct types of synapse could be recognized. A minimum estimate of the total number of synapses on the motor cell is 300, based on their occurrence in reconstructed segments.

Animals↗
PubMed1972

A comparison of chemical and electrical synaptic transmission between single sensory cells and a motoneurone in the central nervous system of the leech.

In leech ganglia, three sensory cells of different modality converge on a motoneurone, where they form chemical and electrical synapses. Each of these synapses behaves in a characteristic manner and the nature of the transmission mechanism has significant functional consequences for the operation of the reflexes. An analysis has been made of the effects of trains of impulses on synaptic transmission through these pathways, using frequencies that correspond to natural firing.1. At the chemical synapse between the nociceptive sensory cell and the motoneurone, two opposing events occur: facilitation and depression. Thus, with trains of impulses, the synaptic potentials first increase in amplitude and then decrease. The two processes could be separated by altering the Mg and Ca content of the bathing fluid. In concentrations of Mg that reduced the amplitude of a single control chemical synaptic potential, pure facilitation occurred during a train. Depression predominated during brief trains in raised concentrations of Ca, although synaptic potentials were initially larger. These results suggest that changes in the amount of transmitter released by each presynaptic action potential can account for the changes observed in chemical synaptic transmission.2. In contrast, electrical transmission between the sensory cell responding to touch and the same motoneurone did not show facilitation or depression. The electrical coupling potential in the motoneurone was relatively constant when the touch cell fired at high or low frequencies in normal Ringer fluid, high Mg, or high Ca fluid.3. Further differences between chemical and electrical synapses were apparent when the preparation was cooled to 4 degrees C. In the cold the latency of chemically evoked synaptic potentials in the motoneurone increased and their amplitude declined drastically with repetitive stimulation, while electrical coupling potentials were unaffected.4. A brief hyperpolarization of the presynaptic cell by injected current produced a marked and prolonged increase in chemically evoked synaptic potentials, but did not influence electrical synaptic transmission.5. The synapses of the sensory cell responding to pressure, which are both chemical and electrical, behaved as expected: the chemical synaptic potentials showed facilitation and depression while electrical transmission remained relatively constant.6. These experiments emphasize the different functional consequences of electrical or chemical synapses in reflex pathways for the transmission of signals that arise as a result of natural sensory stimuli.

Action Potentials↗
PubMed1970

Monosynaptic chemical and electrical connexions between sensory and motor cells in the central nervous system of the leech.

The synaptic connexions that underlie three different segmental shortening reflexes have been traced by recording intracellularly from individual sensory and motor nerve cells in the C.N.S. of the leech. The fourteen sensory cells involved in these reflexes respond specifically to one of three modalities: touch, pressure, or noxious stimuli applied to the skin. All three types of sensory neurone give rise to excitatory synaptic potentials in two large motoneurones. Each of these motor cells provides excitatory innervation to the longitudinal muscle fibres of the opposite side of the segment. The mechanism of synaptic transmission is, however, different for each type of sensory cell.1. An impulse in a sensory cell that responds to touch gives rise to a short-latency depolarizing potential in the large longitudinal motoneurones by way of an electrical synapse. This junction rectifies so that excitation can spread in only one direction (from the sensory to the motor cell), whereas a hyperpolarizing potential can pass only in the opposite direction.2. The synaptic potential evoked in the motoneurone by an action potential in a sensory cell responding to noxious stimuli can be attributed to the action of a chemical transmitter agent and has different properties: the post-synaptic potential arises after a delay of about 2-4 msec, is abolished by high concentrations of Mg, and enhanced by high concentrations of Ca. Several lines of evidence show that this connexion is monosynaptic.3. The synaptic potential following an impulse in a pressure cell is produced by both chemical and electrical synaptic mechanisms. Rectification, similar to that described for the touch cell, also occurs at this electrical synapse.4. One or more impulses in any one of the fourteen mechanoreceptor cells in the ganglion can initiate impulses in the large longitudinal motoneurones to produce a shortening of the segment. The contraction is abolished by blocking impulse initiation in the motoneurones.5. The arborizations of the sensory cells and the motoneurone within the neuropile have been studied histologically after injecting a fluorescent dye. Their processes are intertwined in a highly complex manner so that the sites of the synaptic junctions cannot be determined with the resolutions so far achieved. Nevertheless, taken together the histological and the electrical results support the idea that individual cells are connected in a stereotyped pattern and operate by distinctive mechanisms.6. These findings provide a basis for studying the functional role of chemical and electrical synaptic mechanisms in these pathways.

Action Potentials↗
PubMedDate not supplied

Changes in the dendritic branching of adult mammalian neurones revealed by repeated imaging in situ.

A major obstacle to understanding the mechanism of long-term change in the vertebrate nervous system has been the inability to observe the same nerve cell at different times during the life of an animal. The possibility that changes in neural connectivity underlie the remarkable flexibility of the nervous systems of mammals has therefore not been tested by direct observation. Here, we report studies in which we have visualized the same neurone in the superior cervical ganglion of young adult mice at intervals of up to 33 days. This collection of nerve cells is particularly accessible and therefore well suited to our approach. We find that the dendritic branches of the neurones examined change appreciably over intervals of 2 weeks or more; some branches retract, others elongate and others seem to form de novo. The apparent remodelling of these postsynaptic elements implies that the synaptic connections of these cells normally undergo significant rearrangement beyond what is usually considered to be the developmental period.

Animals↗
PubMedDate not supplied

Structure of the human sensorimotor system. I: Morphology and cytoarchitecture of the central sulcus.

We have studied the morphology of the central sulcus and the cytoarchitecture of the primary sensorimotor cortex in 20 human brains obtained at autopsy. Although the surface appearance of the central sulcus varies greatly from brain to brain (and between hemispheres of individual brains), its deep structure is remarkably consistent. The fundus of the central sulcus is divided into medial and lateral limbs by a complex junction midway between the sagittal and Sylvian fissures. Based on functional imaging studies, this junction appears to be a structural hallmark of the sensorimotor representation of the distal upper extremity. We also identified and measured area 4 (primary motor cortex) and area 3 (primary somatic sensory cortex) in Nissl-stained sections cut orthogonal to the course of the central sulcus. Although the positions of the cytoarchitectonic boundaries in the paracentral lobule showed considerable interindividual variation, the locations of the borders of areas 4 and 3 along the course of the sulcus were similar among the 40 hemispheres examined. In addition to describing more thoroughly this portion of the human cerebral cortex, these observations provide a basis for evaluating lateral symmetry of the human primary sensorimotor cortex.

Cerebral Cortex↗
PubMedDate not supplied

Structure of the human sensorimotor system. II: Lateral symmetry.

We have evaluated the lateral symmetry of the human central sulcus, brainstem and spinal cord using quantitative histological and imaging techniques in specimens from 67 autopsy cases. Our purpose was to determine whether the preferred use of the right hand in the majority of humans is associated with grossly discernible asymmetries of the neural centers devoted to the upper extremities. In the accompanying report, we described a consistent set of morphological features in the depths of the central sulcus that localize the sensorimotor representation of the distal upper extremity. Measurements of the cortical surface in this region, and indeed throughout the entire central sulcus, showed no average lateral asymmetry. Cytoarchitectonic measurements of area 4 and area 3 confirmed this similarity between the left and right hemispheres. The medullary pyramids, which contain the corticospinal tracts, were also symmetrical, as were the cross-sectional areas of white and gray matter in the cervical and lumbar enlargements of the spinal cord. Finally, we found no lateral difference in the size and number of motor neurons in the ventral horns at these levels of the cord. Based on these several observations, we conclude that the preferred use of the right hand in humans occurs without a gross lateral asymmetry of the primary sensorimotor system.

Adult↗