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Biomedical subjects

P M Field

Publications and source records attributed to P M Field.

At least 37 records · Page 2Linked to original sources
PubMed1985

The density of reinnervation of adult rat superior cervical sympathetic ganglionic neurons is limited by the number of available postsynaptic sites.

The adult rat superior cervical ganglion has about 27,000 neurons and is innervated by about 9000 preganglionic axons which make a total of nearly 11 million synapses. Surgical removal of the upper part of the ganglion, reducing the number of neurons to about 20%, causes an overall reduction of the number of synapses to about 30%, but has no effect on the numbers of preganglionic axons. Thus, a 5-fold increase in the axon/neuron ratio causes an increase of only about 50% in the number of synapses per cell. Axotomy followed by regeneration of the preganglionic axons causes no further increase in the number of synapses per cell, even though the average number of synapses per axon is reduced to about one-quarter of the normal. This suggests that the ganglionic neurons can only accept a limited number of synapses, and that in the normal situation there is only possibility for a relatively minor increase before this limit is reached. This study is complementary to a previous one in which the numbers of preganglionic axons were surgically reduced and it was found that, when allowed to regenerate into an entire denervated ganglion, the remaining axons could not increase their numbers of synapses. Thus, in the normal rat superior cervical sympathetic ganglion the total number of synapses is such that while the preganglionic axons are probably expressing close to their full synaptogenic potential, the ganglionic neurons express only about two-thirds of their ability to receive synapses.

Animals↗
PubMed1983

Relative slowness of heterotypic synaptogenesis in the septal nuclei.

The dorsolateral quadrant of the lateral septal nucleus receives a bilateral projection from the fimbria. When the fimbria of one side is cut, the axons of the remaining fimbria take over its synaptic sites preferentially, but when both fimbrias are cut the sites are reinnervated by non-fimbrial axons. To explore the basis of this preference, the present study plots the time courses of the appearance and disappearance of degenerating synapses, and the loss and recovery of non-degenerating synapses after ipsi-, contra- and bi-lateral fimbrial lesions. A preliminary investigation showed that at any time after these three lesions there was no change in the numerical density per unit area of 'control' structures such as shaft synapses (which do not degenerate) and neuronal perikarya (which neither shrink nor degenerate). This indicates that the changes in the numerical density of fimbrial (spine) synapses can be used as a measure of the processes of deafferentation and reinnervation without the danger of the numerical data being distorted by shrinkage. In the sampled area, the ipsilateral fimbrial axons account for about 45% of the synapses and the contralateral fimbrial axons for 25%. The number of degenerating synapses appearing at any one time underestimates the loss of non-degenerating synapses by about one-third, and a photographic simulation of degeneration suggests that a major factor in this discrepancy is the difficulty in recognizing degenerating synapses. Our main finding is that there is a major delay in the rate of removal of degeneration, and in the rate of reinnervation, after bilateral as opposed to unilateral lesions. This delay cannot be accounted for in any simple way by the greater amounts of degeneration. Thus after unilateral lesions, which cause the turnover of 25% (contralateral) or 45% (ipsilateral) of the synapses, 50% of the degeneration is removed in 1-2 days after the peak, whereas after bilateral lesions, which affect 70% of the synapses, it takes 20 days for 50% of the degeneration to be removed. That the synaptic changes after bilateral lesions involve a qualitatively different mechanism is also suggested by the observations of a much greater proportional increase in the multiple synapse index, and a decreased astroglial response.

Afferent Pathways↗
PubMed1983

Neurological mapping using an electron microscope/microprocessor data acquisition system.

Quantitative electron microscopy is used widely in many neurobiological laboratories but can be greatly enhanced by the computer storage, display and analysis of features or boundaries observed under the microscope. An interactive system is described, in which a microprocessor and microcomputer combination controls the data acquisition and display; more detailed analysis can be carried out on a mainframe computer. The performance of this system is examined and it is shown to be a cheap and effective solution, capable of expansion in many directions.

Animals↗
PubMed1980

Synapse formation after injury in the adult rat brain: failure of fimbrial axons to reinnervate the bed nucleus of the stria terminalis.

Selectivity in the reinnervation of denervated postsynaptic sites in the adult rat septal nuclei has been studied by both light and electron microscopic degeneration techniques after lesions of the fimbria and stria terminalis. In the mid-rostrocaudal septum the ventral border of the lateral septal nucleus is coextensive with the dorsal border of the strial bed nucleus. In the normal rat, fimbrial axons establish synapses throughout the lateral septal nucleus of the same side, and also in the dorsal part of the lateral septal nucleus on the opposite side. The stria terminalis establishes synapses in the ipsilateral but not in the contralateral bed nucleus at this level. Both the fimbria and the stria terminalis were completely severed on the left side, and after adequate survival for the removal of all degeneration, the distribution of the remaining fimbria was plotted. Interesting changes were found on the side contralateral to the second lesion, where the fimbria both increases the number of its synaptic terminals within its proper contralateral territory (the dorsal part of the lateral septal nucleus) and also extends its distribution into the ventral part of the lateral septal nucleus--the territory normally reserved for the ipsilateral fimbria. Although completely surrounding the strial bed nucleus, fimbrial axons fail to invade the bed nucleus, and fimbrial terminals are therefore unable to reinnervate denervated strial postsynaptic sites. Since there are no obvious structural barriers between the neuropil of the lateral septal nucleus and that of the strial bed nucleus it is suggested that this failure is most likely to be due either to some biochemical incompatibility between fimbrial axons and strial postsynaptic sites, or to the fact that the fimbrial axons are denied access because some other (unidentified) axonal system forms new presynaptic terminals which effectively pre-empt the sites in the strial bed nucleus.

Animals↗
PubMed1980

Synapse formation after injury in the adult rat brain: preferential reinnervation of denervated fimbrial sites by axons of the contralateral fimbria.

The dorsolateral quadrant of the lateral septal nucleus receives projections from both the ipsilateral and the contralateral fimbria. In the adult rat the effect of fimbrial lesions on synapse formation has been studied by a quantitative electron microscopic analysis of the various types of synapses present, using electron-dense degeneration to identify fimbrial fibre terminals. In this area, the fimbrial axons from both sides together account for about 30% of the total number of synapses and they terminate mainly on dendritic spines. The ipsilateral fimbria forms twice as many synapses as the contralateral fimbria. When one fimbria is cut and time left for the degeneration to be removed, the numbers of synapses are restored to normal levels and the remaining fimbria acquires, on both sides of the septum, a number of synapses equal to the sum of the two individual fimbria, This suggests that the axons of the surviving fimbria have completely reinnervated the denervated postsynaptic sites formerly occupied by the cut fimbria of the other side, effectively excluding non-fimbrial axon terminals, even though the latter constitute the majority (70%) of the synaptic terminals in the region. When both fimbria are cut the numbers of synapses are once again restored to normal levels. However, since there are now no fimbrial axons left, the denervated fimbrial postsynaptic sites must this time have been reinnervated by non-fimbrial axons. Reinnervation by non-fimbrial axons is numerically equally effective in reclaiming the denervated sites, although when compared to the reinnervation by fimbrial axons, the removal of degenerating terminals is somewhat slower, and among the reinnervating terminals there is a much higher incidence of axon terminals making more than one synaptic contact in the plane of section. Thus, fimbrial axons, when present, have the ability to exclude the reinnervation of denervated fimbrial sites by non-fimbrial axons, despite the fact that the latter are both more numerous and also clearly capable of reinnervating those sites when no fimbrial axons are present. Two possible mechanisms are discussed: a spatial preference based on the geometrical arrangements in the neuropil, and a temporal preference based on the relative rates of response of the fimbrial vs the non-fimbrial axons.

Animals↗
PubMed1979

Decentralization of the superior cervical ganglion in neonates impairs the development of the innervation of the iris. A quantitative ultrastructural study.

The superior cervical sympathetic ganglion was decentralized unilaterally by section of the preganglionic chain in 2- or 4-day-old rats. Twenty-one days post-operatively the dilator muscle of the iris was examined electron microscopically. Quantitative estimates demonstrated that there was a reduction both in the numbers of axon bundles and of individual axons in the iris innervated by the deafferented ganglion as compared with its fellow of the contralateral control side.

Animals↗
PubMed1979

Muscarinic receptors in the central nervous system of the rat. I. Technique for autoradiographic localization of the binding of [3H]propylbenzilylcholine mustard and its distribution in the forebrain.

[3H]Propylbenzilylcholine mustard ([3H]PrBCM) is a synthetic, potent muscarinic antagonist, which binds specifically and irreversibly by means of a covalent linkage to muscarinic receptors. Ten micrometer coronal cryostat sections taken through unfixed rat brain at the level of the maximum extent of the caudate nucleus were mounted on glass slides and incubated with 2.4 nM [3H]PrBCM at 30 degrees C for 25 min. They showed a total binding of 3250 pmol/g protein, of which 2130 pmol/g protein was sensitive to pretreatment with 10-6 M atropine. The specific (atropine-sensitive) binding was saturable. Saturation was reached at 15 min, with a rate constant of 1.3 x 106 M-1 sec-1. Binding was unaffected by drugs acting at nicotinic receptors (D-tubocurarine, hexamethonium), or by physostigmine, but was inhibited by muscarinic drugs (pilocarpine, oxotremorine, 3-quinuclidinylbenzilate). Postfixation for 15 min in Carnoy's fixative reduced the specific binding by 10% and the non-specific by 50%. Prefixation (i.e. before incubation with [3H]PrBCM) with any fixatives containing formaldehyde largely prevented specific binding, but a range of concentrations of glutaraldehyde (2% to 0.05%) caused only small reductions in specific binding (e.g. 0.1% glutaraldehyde caused only a 6% reduction). Clear, regionally specific patterns of localization of specific label in light microscope autoradiographs could be obtained from cryostat sections prefixed with 0.1% glutaraldehyde, incubated with 2.4 nM [3H]PrBCM for 15 min at 30 degrees C, and postfixed for 15 min in Carnoy's solution. Of the 105 forebrain areas studied 12 had grain counts between 6 and 9 times the non-specific level and a further 30 had counts 4 to 6 times non-specific. The higher grain counts were in the external plexiform layer of the olfactory bulb, anterior olfactory nucleus, olfactory tubercle, pyriform cortex, stratum radiatum of the hippocampus, stratum moleculare of the dentate gyrus, lateral amygdaloid nucleus, cortico-amygdaloid transition zone, anteroventral thalamic nucleus, hypothalamic supraoptic nucleus, caudate-putamen, nucleus accumbens, and in laminae 3 and 6 of the neocortex (parietal region). There were high grain densities over the choroid plexus the lateral but not the third or fourth ventricles.

Animals↗
PubMed1979

Muscarinic receptors in the central nervous system of the rat. II. Distribution of binding of [3H]propylbenzilylcholine mustard in the midbrain and hindbrain.

The distribution of muscarinic receptors has been studied in the rat midbrain and hindbrain by counting silver grains in light microscope autoradiographs of the specific (atropine-sensitive) binding of [3H]propylbenzilylcholine mustard in cryostat sections. Of the 78 areas studied 6 had grain counts between 6 and 9 times the nonspecific level ("high"), and a further 15 had counts 4-6 times non-specific ("intermediate"). The basilar pontine nuclei and the ventral nuclei of the lateral lemniscus had high counts. Among the cranial nerve motor nuclei the facial and hypoglossal nuclei had high counts and the motor trigeminal nucleus and nucleus ambiguus had medium counts. The interpeduncular nucleus as a whole had low counts but there were two bands of intense staining on each side around the entry zone of the bundles of afferent cholinergic fibres from the habenula. Intermediate levels of binding occurred over the inferior colliculus and the superficial and intermediate grey layers of the superior colliculus. The molecular layer of the vestibulocerebellar vermis was distinctly labelled.

Animals↗
PubMed1979

Muscarinic receptofs in the central nervous system of the rat. III. Postnatal development of binding of [3H]propylbenzilylcholine mustard.

The postnatal development of muscarinic receptors has been studied in 7 selected areas from the brains of 1-17-day-old rats by counting silver grains in light microscope autoradiographs of the specific (atropine-sensitive) binding of [3H]propylbenzilylcholine mustard in cryostat sections. A major part of the adult receptor density is present at 1 day of age, a time when only a small fraction of the adult number of synapses has yet been formed. Of the areas studied the hypoglossal nucleus is the most precocious in muscarinic receptor development, and the dentate gyrus the latest (associated with the late development of the dentate granule cells). The pattern of receptor distribution changes with development. The caudate-putamen first develops receptor in patches, beginning at the lateral (ventricular) surface. The pontine nuclei develop receptor in a medial to lateral sequence. The maturation of the adult laminar pattern of the olfactory bulb depends on the alignment of cells (especially the mitral cells). The neocortex initially has uniform labelling throughout its depth, and later the labelling in layer 4 becomes relatively less dense (probably associated with the ingrowth of afferent fibres). The hippocampal formation first develops receptor evenly over the pyramidal cell dendrites; later receptor appears over the newly formed dentate stratum moleculare and becomes much reduced over the hippocampal stratum lucidum and stratum lacunosum-moleculare (probably associated with the ingrowth of afferent fibres from the dentate gyrus and entorhinal area). In the cerebellum muscarinic receptor is found only in the lobules which receive the primary vestibular afferents. In the neonate it is present in the granular layer, but this later disappears and is replaced by the adult pattern of labelling in the molecular layer.

Aging↗
PubMed1979

Muscarinic receptors in the central nervous system of the rat. IV. A comparison of the effects of axotomy and deafferentation on the binding of [3H]propylbenzilylcholine mustard and associated synaptic changes in the hypoglossal and pontine nuclei.

The reaction of axotomy has been studied in the rat hypoglossal nucleus by quantitative electron microscopical counts of numbers of synapses and by changes in muscarinic receptors assessed by counting silver grains in light microscope autoradiographs of the specific (atropine-sensitive) binding of [3H]propylbenzilylcholine mustard in cryostat sections. For the first 5 days after unilateral peripheral hypoglossal nerve axotomy the muscarinic ligand binding falls to 50% of control levels and then shows no further fall for up to 30 days. Synapse numbers decrease progressively over the first 10 days after operation, by which time they reach 50% of normal. Thus receptor changes reach completion at a time when synapse loss is still continuing. Later, both muscarinic ligand binding and synapse numbers recover to an extent which depends at least in part on the effectiveness of the peripheral nerve regeneration, suggesting that both the receptor and synapse changes may be dependent upon neuromuscular contacts. The reactions of muscarinic receptors to axotomy and deafferentation have been studied in the rat basilar pontine nuclei. Cerebellectomy, which causes axotomy of the pontine neurones and also removes their postsynaptic targets (the granule cells), causes no change in pontine muscarinic receptor over the first week after operation. This differs from the rapid fall in hypoglossal muscarinic receptors induced by axotomy. At longer survivals after cerebellectomy there is a partial loss of pontine muscarinic receptors associated with atrophy of the pontine neurones. Destruction of the neocortical afferents causes a loss of at least half of the synapses in the pontine neuropil. However, the light microscopic autoradiographic study revealed no obvious changes in the dentisy or distribution of the pontine muscarinic receptors from 4 days to more than 6 months after operation.

Afferent Pathways↗
PubMed1978

Electron microscope autoradiographic evidence for specific transneuronal transport in the mouse accessory olfactory bulb.

The distribution of radioactive material was examined autoradiographically 8 h after application of [3H] proline to the vomeronasal organ in mice. Labelled material was transported along the axons of the vomeronasal nerves to their terminals in the glomerular layer of the accessory olfactory bulb (AOB). A lesser but consistent amount of radioactivity was found in the external plexiform layer (EPL) of the AOB. Electron microscopic autoradiography was used to determine which of the components of the EPL contained this labelled material. The method of proportional grain counts showed that the highest concentration of silver grains lay over the mitral cell dendrites, which are the elements immediately postsynaptic to the vomeronasal nerve axons. However, a fairly high proportion of grains also lay over the peripheral processes of granule cells. By application of a method of 'crossfire analysis' (which is explained in detail) it was possible to show that the observed grain distribution is best explained by the assumption that the radioactive material is confined to mitral cells, and the labelling over granule cell processes is due to crossfire from these sources. Im one animal at 5 days after [3H]proline administration label was found to have extended from mitral cells to granule cells, suggesting that the transsynaptically transported radioactive material, which was confined to the mitral cells at 8 h, may have become further redistributed at longer survivals. In a control experiment, [3H]proline was applied directly to the surface of the AOB. This gave rise to a completely different distribution of radioactivity in the EPL: radioactive material was present in all tissue components.

Animals↗
PubMed1976

A quantitative comparison of the formation of synapses in the rat superior cervical sympathetic ganglion by its own and by foreign nerve fibres.

The rat superior cervical sympathetic ganglion (SCG) has about 36,000 neurones in a volume of about 1 cu.mm. There are about 8.8 X 10(6) synapses, and 6000-9000 preganglionic axons. Section of the preganglionic chain causes a loss of 93% of the synapses. In the denervated SCG there are 0.6 X 10(6) remaining ('intrinsic') synapses, and a proportion of the synaptic sites are identifiable as vacated synaptic thickenings (3 X 10(6) per SCG, as compared with 0.5 X 10(6) in the normal intact SCG). After deducting the intrinsic synapses, this indicates that each preganglionic axon forms about 1100 (900-1400) synapses. After freezing the preganglionic chain, subsequent axonal regeneration restores synapse numbers to 85% of normal (7.5 X 10(6) synapses per SCG). After anastomotic repair by suture of the cut ends of the preganglionic chain (a necessary control for the foreign nerve anastomoses), the SCG contains only 60% of the normal complement of synapses (5.2 X 10(6) synapses per SCG). The results of this anastomosis are very variable. However, in individual ganglia the numbers of synapses are directly correlated with the numbers of axons which reach the SCG. After deducting the intrinsic synapses it can be calculated that each axon forms about 700 synapses. This is probably an underestimation of the numbers which would be achieved at longer survival times. After anastomosis of the vagal nerve into the denervated SCG there are about 4.4 X 10(6) synapses per SCG. Morphologically the majority have axon terminals with large dense cored vesicles, and it is likely that these belong to the axons of the parasympathetic preganglionic neurones in the dorsal motor nucleus of the vagus. A smaller population of axon terminals are devoid of large dense cored vesicles; their origin is unknown. The dorsal motor nucleus of the vagus has between 1000 and 2000 neurones. After deducting the intrinsic synapses, this indicates that each axon may form up to 1900-3800 synapses. To the extent that other, unidentified vagal fibres also contribute to the synapses found after this anastomosis, this figure is an overestimate. After anastomosis of the hypoglossal nerve into the denervated SCG, there are 1.5 X 10(6) synapses per SCG. A morphologically distinctive type of axon terminal is found, and it is argued that this may belong to a special category of skeletomotor neurones located in the caudoventral part of the hypoglossal nucleus and distinguished by pseudocholinesterase staining. There are about 600 of these neurones, which would indicate that they form about 1500 synapses per axon (after deducting the numbers of intrinsic synapses). The majority of the hypoglossal neurones do not form intraganglionic synapses; this suggests that although the possession of a cholinergic mechanism may be necessary for axons to be able to form ganglionic synapses, it is not in itself sufficient. For each of the types of anastomosis, the numbers of vacated thickenings are inversely proportional to the numbers of synapses...

Acetyltransferases↗
PubMed1975

Retrograde transport of horseradish peroxidase in the magnocellular neurosecretory system of the rat.

Horseradish peroxidase (HRP) was injected into the pituitary in adult rats. After 2-3 days, the neurones of the supraoptic nuclei, the magnocellular parts of the paraventricular nuclei, and the various accessory neurosecretory hypothalamic nuclei showed accumulation of HRP. The HRP reaction product consisted of fine, discrete cytoplasmic granules, and in electron micrographys it was seen to be located in the lysosome-like dense bodies of 0.4-0.6 mum diameter which are normally present in the cytoplasm of the neurosecretory neuron.es. Very little reaction product was found in the neurosecretory axons. Reaction product was also found in the hypothalamic arcuate nuclei. This was the result of an endogenous peroxidase-like activity, since it occurred in control animals which had not received HRP. This endogenous reaction product is non-neuronal. Morphologically, it takes the form of distinctive clusters of coarse granules which are seen in electron micrographs to be characteristic angular bodies of 0.7-1.0 mum diameter located in the cytoplasm of astrocytes or their processes.

Animals↗
PubMed1975

Cellular localization of tyrosine hydroxylase by immunohistochemistry.

The enzyme tyrosine hydroxylase (TH) was immunohistochemically localized by the peroxidase-antiperoxidase method in rat to chromaffin cells of the adrenal medulla, large neurons and small darkly staining cells of the superior cervical ganglia and noradrenergic and dopaminergic neurons in brain. As compared with the conjugated peroxidase or immunofluorescence techniques, the peroxidase-antiperoxidase method gave the most selective and specific cytoplasmic localization of TH antisera in every tissue examined. The peroxidase staining with the TH antisera was more intense in dopaminergic than in noradrenergic neurons of the central nervous system. While TH was visualized in cell bodies of both dopaminergic and noradrenergic neurons, it could only be detected in axons and terminals in the dopaminergic system. The perikarya of noradrenergic neurons could be distinguished from dopaminergic neurons by the immunohistochemical demonstration of the enzyme dopamine-beta-hydroxylase only in the former.

Adrenal Medulla↗