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D Purves

Publications and source records attributed to D Purves.

At least 91 records · Page 5Linked to original sources

Regulation of synaptic connections in the rabbit ciliary ganglion.

One of the intriguing questions about the establishment of synaptic connections is how appropriate numbers of different axons come to innervate each target neuron. A reorganization of connections in early postnatal life appears to be an important aspect of this process, since many of the axons terminals that initially innervate target cells are subsequently lost. The rabbit ciliary ganglion is a remarkably simple neural ensemble in which to examine this rearrangement of developing synaptic connections. Using this system we have found that a reduction in the number of axons innervating each cell occurs without any change in the number of ciliary ganglion cells or preganglionic neurons; therefore the rearrangement is not based on cell death. The number of different axons that ultimately innervate each cell is, however, influenced in some way by the geometry of individual target neurons. Thus, mature ganglion cells that lack dendrites are generally innervated by a single axon, while neurons with increasingly complex dendritic arbors receive innervation from a commensurate number of different axons. At birth, on the other hand, neurons with or without dendritic processes receive about the same number of preganglionic inputs. These results suggest that the geometry of the target cell influences the competitive interaction between different axons innervating the same neuron. Indeed, an important function of dendrites may be to regulate the number of axons that innervate each nerve cell.

Aging↗

Re-innervation of ganglia transplanted to the neck from different levels of the guinea-pig sympathetic chain.

Thoracic and lumbar sympathetic ganglia from donor guinea-pigs were transplanted to the bed of an excised superior cervical ganglion in host animals. Homotopic transplants of superior cervical ganglia served as controls. In this way the same set of preganglionic axons (the cervical sympathetic trunk) was confronted with ganglia from different levels of the sympathetic chain. Re-innervation of the transplants was studied after 3-5 months. 1. Neurones in ganglia transplanted from different levels of the sympathetic chain were re-innervated to about the same over-all degree by the preganglionic axons of the host's cervical sympathetic trunk. Thus, the mean amplitude of post-synaptic potentials, the estimated number of innervating axons, and the number of spinal segments providing innervation to each neurone were similar in transplanted thoracic, lumbar and superior cervical ganglion cells. 2. Neurones in transplanted mid-thoracic ganglia, however, were re-innervated more frequently, and more strongly, by axons arising from more caudal thoracic segments than neurones in transplanted superior cervical ganglia. Stimulation of axons arising from the fourth thoracic spinal segment (T4), for example, elicited post-synaptic potentials that on average were twice as large in transplanted fifth thoracic ganglion cells as in transplanted superior cervical ganglion cells; conversely, axons arising from T1 re-innervated neurones in the superior cervical ganglion about 2-3 times more effectively than fifth thoracic ganglion cells. This difference in the re-innervation of the fifth thoracic and the superior cervical ganglion is in the same direction as (although less pronounced than) the normal difference in the segmental innervation of these ganglia. 3. Transplanted lumbar ganglia were also re-innervated more effectively by relatively caudal segments compared to re-innervated cervical ganglia, but this difference was no greater than that observed for transplanted thoracic ganglia. 4. We conclude that preganglionic axons can distinguish (or be distinguished by) ganglia derived from different levels of the sympathetic chain. Our findings are consistent with the view that ganglion cells have some permanent property that biases the innervation they receive.

Animals↗

Post-natal reduction of neural unit size in the rabbit ciliary ganglion.

We have studied the innervation of adult and neonatal ciliary ganglia in the rabbit to determine the average number of ganglion cells innervated by each preganglionic neurone at different stages of development. 1. The adult ciliary ganglion comprises about 380 ganglion cells which are innervated by about forty preganglionic neurones. 2. Ciliary ganglion cells in adult rabbits are on average innervated by 2.2 different axons; in contrast, neonatal ganglion cells are on average innervated by 4.6 different axons. The transition to the adult pattern of innervation occurs gradually during the first few post-natal weeks. 3. The numbers of ganglion cells and preganglionic neurones do not change appreciably after birth. Accordingly, the loss of some innervation to individual neurones during post-natal development indicates that each preganglionic axon innervates progressively fewer ciliary ganglion cells. 4. The number of synaptic boutons found in ganglia at birth, however, is less than the number of synaptic boutons found in adult ganglia. 5. We conclude that synaptic connexions in this ganglion age gradually rearranged in early post-natal life such that each preganglionic neurone focuses an increasing number of synaptic contacts on a progressively smaller subset of the ganglion cell population.

Aging↗

The relation of postsynaptic geometry to the number of presynaptic axons that innervate autonomic ganglion cells.

We have studied the shape of rabbit ciliary ganglion cells in relation to the number of axons that innervate each neuron. Adult ganglion cells receive synapses from one to seven different preganglionic axons. Some neurons lack dendrites altogether, whereas others have complex arborizations of up to eight primary dendrites. The neurons that receive all of their synaptic contacts from a single preganglionic axon usually have no dendrites; on the other hand, multiply innervated ganglion cells receive synapses from a number of different axons that increases in proportion to the number of primary dendrites that they possess. A possible explanation of these results is that individual ciliary ganglion cells comprise a number of separate spatial domains, each of which is largely constrained to receive innervation from a single preganglionic axon.

Animals↗

Segmental organization of sympathetic preganglionic neurons in the mammalian spinal cord.

We have used retrograde transport of horseradish peroxidase to determine the distribution of the preganglionic cell bodies whose axons join particular rami of the thoracic spinal cord in a series of guinea pigs, and in a small number of hamsters and cats. In contrast to other recent studies, our results show that the neurons sending axons to a ramus are confined to a single segment at the corresponding spinal level. This segmental organization supports the idea that the rostro-caudal position of preganglionic cell bodies is one determinant of selective synapse formation between preganglionic axons and sympathetic ganglion cells.

Adrenergic Fibers↗

Innervation of sympathetic neurones in the guinea-pig thoracic chain.

We have investigated the organization of the guinea-pig thoracic chain by studying the innervation of the stellate and fifth thoracic sympathetic ganglia with intracellular recording. 1. These ganglia receive preganglionic innervation from different but overlapping sets of spinal cord segments: the stellate ganglion is innervated by preganglionic axons from spinal segments more rostral than those supplying the fifth thoracic ganglion, but somewhat more caudal than those innervating the superior cervical ganglion. 2. Individual thoracic ganglion cells are innervated by only some of the spinal segments that supply each ganglion as a whole. In general, the subset of spinal segments innervating a ganglion cell is contiguous; one of these segments provides the strongest innervation, with progressively weaker innervation arising from spinal levels adjacent to the dominant one. This selective pattern of innervation is similar to that in the superior cervical ganglion (Njå & Purves, 1977 a). 3. Preganglionic axons frequently innervate neurones in more than one ganglion. 4. Although neurones innervated by the same spinal cord segments are found in both the stellate and the fifth thoracic ganglion, as well as in the superior cervical, the number of ganglion cells receiving innervation from particular spinal segments is different in each ganglion. Moreover, neurones dominated by the same segment but located in different ganglia receive somewhat different average innervation from adjacent segments as a function of the ganglion in which they reside. 5. These results indicate that neurones in the thoracic chain ganglia, as those in the superior cervical ganglion, are selectively innervated by particular spinal cord segments. We suggest that the different average innervation of sympathetic ganglia reflects at least two related factors: the selective qualities of their constituent neurones, and the availability of different preganglionic axons to each ganglion.

Animals↗

The elimination of redundant preganglionic innervation to hamster sympathetic ganglion cells in early post-natal life.

The superior cervical ganglion of adult and neonated hamsters has been studied with intracellular recording. 1. Neurones in adult hamster ganglia are innervated by an average of 6-7 preganglionic axons. During the first week of post-natal life, however, these cells are innervated by at least eleven to twelve axons. Ganglion cells in animals 2-3 weeks old are innervated to an intermediate degree, indicating that these neurones lose a substantial portion of their initial synaptic contacts during the first weeks after birth. 2. The over-all innervation of the superior cervical ganglion in adult hamsters arises from thoracic segments T1-T5; no additional segments contribute significantly to the innervation of neonatal ganglia. 3. The average number of segments innervating each adult ganglion cell is 2 . 8 compared to 3 . 7 segments innervating neonatal neurones. Throughout post-natal development the innervation of individual neurones arises from a contiguous subset of the spinal segments that innervate the ganglion as a whole. 4. We conclude that the elimination of redundant innervatin in early life is not limited to those nerve and muscle cells contacted by a sigle axon in maturity, but also occurs in sympathetic ganglia where adult neurones remain multiply innervated. Moreover, the loss of some synaptic contacts during development refines the selective innervation of individual neurones.

Animals↗

On the purpose of selective innervation of guinea-pig superior cervical ganglion cells.

Preganglionic axons arising from different levels of the mammalian spinal cord make preferential connexions with different classes of superior cervical ganglion cells (Langley, 1892, 1900; Njå & Purves, 1977a). For example, preganglionic axons from the first thoracic segment (T1) make relatively strong connexions with ganglion cells activating end-organs of the eye; conversely, axons arising from T4 selectively innervate ganglion cells projecting to the ear. In the present work we have asked whether this selectivity reflects the function of the pre- and post-synaptic cells, and aspect of their respective positions, or some other criterion. 1. End-organs with different functions at the same locus (the eye) respond to stimulation of the same ventral roots; end-organs of a single modality (hairs or blood vessels) at different positions, however, tend to be activated by different spinal segments. Thus the segmental innervation of ganglion cells is correlated with the position rather than the function of post-ganglionic targets. 2. The role of target position in ganglion cell innervation was examined directly by recording from neurones sending axons to different destinations. Superior cervical ganglion cells running dorso-medially in a spinal nerve receive, on average, innervation from more caudal segments than cells projecting ventro-laterally. 3. These selective connexions do not depend on intraganglionic cell position: neurones located at different points along the major axes of the superior cervical ganglion receive, on average, the same segmental innervation. In accord with this observation, retrogradely labelled neurones innervating a particular target such as the eye or ear are widely and randomly distributed within a large portion of the ganglion. Thus the importance of post-ganglionic target position in ganglion cell innervation is not simply a reflexion of ganglionic topography. 4. We conclude that one purpose of the selective connexions in the superior cervical ganglion is to bring together preganglionic axons arising from different levels of the spinal cord and ganglion cells whose axons innervate particular regions of the superior cervical territory.

Action Potentials↗

The effects of post-ganglionic axotomy on selective synaptic connexions in the superior cervical ganglion of the guinea-pig.

Stimulation of preganglionic axons arising from different levels of the thoracic spinal cord causes different effects on end-organs supplied by the superior cervical ganglion (Langley, 1892; Nja & Purves, 1977a; Lichtman, Purves & Yip, 1979). For example, stimulation of the first thoracic ventral root (T1) causes pupillary dilatation and widening of the palpebral fissure; stimulation of T4, on the other hand, has little effect on the eye, even though axons arising from this level innervate about as many superior cervical ganglion cells as those from T1. Thus ganglion cell innervation is selective. (1) Three months after crushing the major post-ganglionic branches of the superior cervical ganglion this differential effectiveness is lost: T1 and T4 stimulation have approximately equal effects on the end-organs of the eye. (2) In normal animals, the cellular counterpart of selective end-organ effects is the innervation of each ganglion cell by a contiguous subset of the spinal segments that innervate the ganglion as a whole. One of these segments is usually dominant, the strength of innervation from adjacent segments falling off as a function of distance from the dominant one (Nja & Purves, 1977a). Intracellular recordings from ganglion cells 3 months after post-ganglionic axotomy showed that this selective pattern is re-established. (3) Since the innervation of ganglion cells appears normal, the abnormal end-organ responses after post-ganglionic axotomy suggest that ganglion cell axons are not limited to their original targets during peripheral re-innervation. This suggestion is supported by the finding that ganglion cells sending axons to different peripheral destinations via the second and third cervical spinal nerves were no longer distinguishable on the basis of their segmented inputs 3 months after post-ganglionic axotomy. (4) Similar results were obtained when the preganglionic cervical trunk was cut at the same time as the post-ganglionic axons were crushed; the pattern of end-organ responses was abnormal, whereas individual ganglion cells were re-innervated according to the rules of contiguity and segmental dominance. (5) These results indicate that ganglion cells do not undergo a compensatory change in the segmental innervation they receive when their axons regenerate to targets different from, or in addition to those they originally innervated, even when an entirely new set of ganglionic connexions is formed. This suggests that ganglion cells, or some aspect of their immediate environment, possess a permanent label that determines the segmental innervation they receive.

Animals↗

Specificity of initial synaptic contacts made on guinea-pig superior cervical ganglion cells during regeneration of the cervical sympathetic trunk.

1. Largely appropriate synaptic connexions are formed with neurones in the superior cervical ganglion at long intervals after interruption of the preganglionic nerve. In the present study we have assessed the accuracy of connexions during the early stages of re-innervation by observing end-organ responses to ventral root stimulation in vivo, and by recording intracellularly from ganglion cells during ventral root stimulation in isolated preparations. 2. Appropriate, but weak, end-organ responses were elicited by stimulation of the first and fourth thoracic ventral roots (T1 and T4) 15--30 days after freezing the cervical sympathetic trunk. 3. Intracellular recordings from ganglion cells during stimulation of the ventral roots C8--T7 in vitro showed that synaptic contacts are first re-established 8--11 days after freezing the preganglionic nerve. The proportion of re-innervated cells, and the strength of innervation of individual neurones, increased rapidly for up to about 3 months after nerve injury, but showed little change thereafter. Innervation remained weaker than normal even after 6 months. 4. Patterns of segmental innervation recorded intracellularly during the early stages of regeneration were similar to, but more restricted than normal. Even 13--19 days after interruption of the preganglionic nerve, neurones re-innervated by more than one spinal cord segment tended to be innervated by a contiguous subset of the spinal segments which contribute innervation to the ganglion. The incidence of neurones receiving innervation from a discontinuous segmental subset was about the same at early and late stages or re-innervation. 5. Throughout the course of nerve regeneration, re-innervated neurones tended to receive dominant synaptic input from axons arising at a particular spinal level, as do normal cells, with adjacent segments contributing a synaptic influence that diminished as a function of distance from the dominant segment. 6. The results of these experiments argue against the initial formation of imprecise connexions with subsequent retention of appropriate contacts and a loss of inappropriate ones. Rather our findings suggest that the re-innervation of ganglion cells proceeds by a gradual accumulation of synaptic connexions which are, from the outset, appropriate.

Animals↗

The effects of nerve growth factor and its antiserum on synapses in the superior cervical ganglion of the guinea-pig.

1. The effects of nerve growth factor (NGF) and its antiserum on synapses in the superior cervical ganglion of the guinea-pig have been examined by intracellular recording and electron microscopy. 2. Exogenous NGF, supplied locally from a silicone rubber pellet implanted near ganglia for 4-7 days, had little effect on either the function or the number of ganglionic synapses. 3. However, the depression of synaptic transmission and loss of synaptic contacts on ganglion cells which follow post-ganglionic axotomy were diminished by about 50% in the presence of exogenous NGF. 4. Other post-axotomy changes such as the development of subthreshold regenerative responses in neuronal processes, the appearance of ultrastructurally abnormal neuronal profiles suggesting rapid membrane turnover, and the cytoplasmic and nuclear changes characteristic of "chromatolysis", were also largely prevented by exogenous NGF. 5. Systemic treatment of neonatal and young adult guinea-pigs with antiserum to NGF for 4-5 days caused depression of intracellularly recorded synaptic responses within 5-8 days of the end of antiserum administration. Synapse counts in electron microscopical sections from these ganglia showed only about half as many contacts as in control ganglia from animals receiving normal rabbit serum. 6. These findings suggest that the loss of synapses from sympathetic neurones which follows axotomy results from a reduction in the amount of NGF supplied to ganglion cells. A corollary is that, among other biological roles, NGF is required by peripheral sympathetic neurones to maintain the synapses they receive.

Animals↗

Specific innervation of guinea-pig superior cervical ganglion cells by preganglionic fibres arising from different levels of the spinal cord.

1. The synaptic contribution of preganglionic nerve fibres arising from the last cervical (C8) and the first seven thoracic spinal cord segments (T1-T7) to neurones of the guinea-pig superior cervical ganglion has been studied by means of intracellular recording during ventral root stimulation in vitro. 2. The majority of neurones received innervation from the middle segments (T2 and T3) of the length of spinal cord from which preganglionic fibres derive; an intermediate number of ganglion cells were innervated by fibres from the segments adjacent to these (T1, T4, and T5), and relatively few neurones by fibres from the most rostral and caudal segments supplying innervation to the ganglion (C8, T6 and T7). 3. Each neurone received preganglionic terminals from multiple thoracic segments (range 1-7, mean = 4-0). The estimated minimum number of preganglionic fibres contacting each neurone was 10, on average. 4. As a rule, the spinal segments innervating a neurone were contiguous. Thus we rarely encountered neurones innervated by segments located both rostrally and caudally to a segment which failed to provide innervation. 5. Neurones tended to be innervated predominantly by axons arising from a single spinal segment, with adjacent segments contributing a synaptic influence that diminished as a function of their distance from the dominant segment. All segments provided dominant innervation to at least some neurones. 6. Stimulating the ventral roots of C8-T7 in vivo showed that the axons arising from each segment produced a characteristic pattern of peripheral effects. Thus different populations of neurones in the superior cervical ganglion of the guinea-pig are innervated by preganglionic axons from different levels of the spinal cord, as originally suggested by Langley (1892) for the cat, dog, and rabbit. 7. On the basis of our in vitro studies we conclude that underlying the specificity of innervation of neurones of the superior cervical ganglion that can be inferred from in vivo experiments is a tendency for individual neurones to be innervated in a systematically graded fashion by a contiguous subset of the eight spinal segments which provide innervation to the ganglion.

Action Potentials↗

Re-innervation of guinea-pig superior cervical ganglion cells by preganglionic fibres arising from different levels of the spinal cord.

1. The ability of preganglionic axons to re-establish their normal pattern of synaptic connexions with superior cervical ganglion cells has been studied after section of the cervical sympathetic trunk.2. In vivo stimulation of the last cervical (C8) and the first seven thoracic ventral roots (T1-T7) 3-4 months after section of the trunk produced end-organ responses similar to those observed in normal animals.3. The pattern of innervation of individual neurones, determined by intracellular recording of synaptic potentials 4-9 months after cutting the sympathetic trunk, was also similar to that observed in normal neurones. Both normal and re-innervated ganglion cells were contacted by pre-ganglionic axons arising from C8 to T7, and each neurone was usually innervated by a contiguous subset of these segments.4. Re-innervated neurones, as normal cells, were typically dominated by the innervation from a particular spinal cord segment, with the adjacent segments contributing a synaptic influence that decreased as a function of distance from the dominant segment. This was true whether the amplitude of the post-synaptic potential, or the estimated number of contributing axons, was used as the criterion of segmental dominance.5. Re-innervated neurones, however, showed some abnormalities. The average number of ventral roots contributing innervation to each neurone was reduced from 4.1 to 3.0, and discontinuities in the sequence of innervating segments were more frequent than in normal neurones. Moreover, fewer preganglionic axons contacted each neurone after regeneration.6. A further difference between normal and re-innervated neurones during the period covered by these experiments was that axons from the more caudal spinal cord segments were less successful in re-establishing contacts with ganglion cells than those from the rostral segments. The more caudal the position of the preganglionic neurones, the more pronounced was this relative deficiency.7. Although anomalies of ganglion cell innervation were apparent, the basis for the restoration of normal functional effects appears to be the re-establishment of a pattern of innervation of individual neurones similar to that observed in normal ganglia.

Action Potentials↗

Synaptic organization and acetylcholine sensitivity of multiply innervated autonomic ganglion cells.

The principal cells of the mudpuppy cardiac ganglion receive synapses from three sources: vagal axons, interneurons and axon collaterals from other principal cells. The simplicity of the structural organization and the visual clarity in the living preparation provide favorable conditions for examining the function of these synapses and how different classes of synapses on the same cell influence its function. We have studied the sensitivity of the principal cells to iontophoretically applied acetylcholine--the transmitter at synapses made by the vagal axons and by postganglionic axon collaterals from other principal cells. In normal ganglia, the ACh sensitivity on the cell surface is highest at the region of synapses. Partial denervation, produced by severing the vagus nerves, results in an increased ACh sensitivity in nonsynaptic areas but does not appear to affect synaptic transmission at the remaining synapses.

Acetylcholine↗