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Biomedical subjects

E Pannese

Publications and source records attributed to E Pannese.

At least 19 recordsLinked to original sources

Satellite cell reactions to axon injury of sensory ganglion neurons: increase in number of gap junctions and formation of bridges connecting previously separate perineuronal sheaths.

This study investigated satellite cell changes in mouse L4 and L5 spinal ganglia 14 days after unilateral transection of sciatic and saphenous nerves. The ganglia were studied under the electron microscope in single and serial sections, and by dye injection. Satellite cell responses to axon injury of the neurons with which they are associated included the formation of bridges connecting previously separate perineuronal sheaths and the formation of new gap junctions, resulting in more extensive cell coupling. Some possible consequences of these satellite cell reactions are briefly discussed.

Animals↗

Glial cell plasticity in sensory ganglia induced by nerve damage.

Numerous studies have been done on the effect of nerve injury on neurons of sensory ganglia but little is known about the contribution of satellite glial cells (SCs) in these ganglia to post-injury events. We investigated cell-to-cell coupling and ultrastructure of SCs in mouse dorsal root ganglia after nerve injury (axotomy). Under control conditions SCs were mutually coupled, but mainly to other SCs around a given neuron. After axotomy SCs became extensively coupled to SCs that enveloped other neurons, apparently by gap junctions. Serial section electron microscopy showed that after axotomy SC sheaths enveloping neighboring neurons formed connections with each other. Such connections were absent in control ganglia. The number of gap junctions between SCs increased 6.5-fold after axotomy. We propose that axotomy induces growth of perineuronal SC sheaths, leading to contacts between SCs enveloping adjacent neurons and to formation of new gap junctions between SCs. These changes may be an important mode of glial plasticity and can contribute to neuropathic pain.

Animals↗

Quantitative changes in mitochondria of spinal ganglion neurons in aged rabbits.

Within the context of our research on the age-related structural changes in spinal ganglia, we studied the mitochondria of the neuronal perikaryon in the spinal ganglia of 12-, 42-, and 79-month-old rabbits. Both the volume of the perikaryon and the total mitochondrial mass within the perikaryon increased significantly passing from young adult to old animals. Hence, there is no net loss of mitochondria in these neurons with age. Since, however, the volume of the perikaryon increased by more than 63% while the total mitochondrial mass within the perikaryon increased by only 18%, the mean percentage of perikaryal volume occupied by mitochondria decreased with age. This decrease is only in very minor part a consequence of lipofuscin accumulation, so that the ratio between the total mitochondrial mass and the functionally active volume of cytoplasm decreased with age. Possible causes of this decrease are discussed briefly. Moreover, while the mitochondrial structure did not change, mitochondrial size increased with age. Finally, in each of the three age groups both the mean percentage volume of mitochondria and the mean mitochondrial size were very similar in large light and in small dark neurons.

Age Factors↗

Changes with age in the Golgi apparatus of rabbit spinal ganglion neurons.

We studied the Golgi apparatus in spinal ganglion neurons of rabbits aged 12, 42 and 79 months. We found no structural changes, no indications of fragmentation, no indications of peripheral displacement affecting this organelle with advancing age. The volume of the perikaryon increased significantly with age, whereas the total volume of the Golgi apparatus remained essentially constant. Hence the mean percentage of perikaryal volume occupied by the Golgi apparatus decreased with age. This decrease was only in very minor part a consequence of lipofuscin accumulation, so that the ratio between the total volume of the Golgi apparatus and the functionally active volume of cytoplasm decreased with age. This decrease could be related to the reduced neuronal metabolism that occurs during ageing. It is possible that the delivery to the axon of newly synthesized proteins destined for fast transport is one of the Golgi apparatus roles that decrease with age. Finally, we found that the age-related quantitative changes in the Golgi apparatus did not differ between large light and small dark neurons.

Aging↗

Decrease in the nucleo-cytoplasmic volume ratio of rabbit spinal ganglion neurons with age.

Nuclear and perikaryal volumes, as well as the mean percentage of perikaryal volume occupied by lipofuscin, were estimated in spinal ganglion neurons of 12, 42 and 79 month-old rabbits. The volume ratio between nucleus and perikaryal cytoplasm (N/C volume ratio) decreased progressively and significantly with increasing age. This was not a consequence of the perikaryal enlargement due to the age-related lipofuscin accumulation since the same result was obtained when the volume occupied by lipofuscin was subtracted from the perikaryal volume. The decrease in the N/C volume ratio may depend on an age-related loss of DNA, a cytoplasmic influence on nuclear size, or other unknown causes.

Aging↗

The perikaryal surface of spinal ganglion neurons: differences between domains in contact with satellite cells and in contact with the extracellular matrix.

The perikaryal surface of spinal ganglion neurons undergoes dynamic changes throughout life. In particular, numerous slender projections develop and retract continuously from this surface. We showed previously that the outgrowth of these projections, while an intrinsic property of spinal ganglion neurons, is also influenced by the surrounding microenvironment. Since the latter consists of satellite cells and the extracellular matrix, we sought to determine the relative contributions of each of these components to the outgrowth of perikaryal projections. To this end, we took advantage of a little known characteristic of the satellite cell sheaths: in the rabbit, these sheaths can exhibit gaps that leave the nerve cell body surface directly exposed to the extracellular matrix. We compared the surface domains covered by satellite cells with those in direct contact with the extracellular matrix. We found that the perikaryal projections are abundant in the former domains but are absent in the latter. We also found that the perineuronal extracellular matrix of rabbit spinal ganglia contains laminin and fibronectin, two glycoproteins that have been reported to promote the growth of axonal processes from sensory ganglion neurons. Laminin and fibronectin were also present at the level of the gaps in the satellite cell sheath. These results: (1) provide additional evidence that environmental factors influence the outgrowth of perikaryal projections from spinal ganglion neurons; (2) suggest that satellite cells permit the outgrowth of these projections; (3) suggest that in the spinal ganglia of adult rabbits the perineuronal extracellular matrix is not in itself able to promote the outgrowth of these projections. This study provides a further example of the influence that supporting neuroglial cells have on sensory ganglion neurons.

Animals↗

The Golgi Stain: invention, diffusion and impact on neurosciences.

The black reaction, invented in 1873 by Camillo Golgi (1843-1926, was the first technique to reveal neurons in their entirety, i.e. with all their processes. This important development passed unnoticed at first and only received wide international attention after a long delay. The Golgi stain was widely employed for almost thirty years and was directly responsible for major advances in our knowledge of the microscopic anatomy of the nervous system, as well as in other fields of study. In the hands of other researchers, the black reaction provided vital evidence that helped to establish the neuron theory. The Golgi stain was almost forgotten in the period between the two World Wars, but the introduction of the electron microscope to neurocytological resarch revived its use around the middle of the twentieth century. Today, the black reaction is still used extensively not only in combination with electron microscopy, but also as an autonomous technique in studies on the evolution, ontogeny, and organization of the nervous system.

Cell Biology↗

Amount and distribution of lipofuscin in nerve and satellite cells from spinal ganglia of young adult and aged rabbits.

The quantitative aspects of the age-related accumulation of lipofuscin were studied in the rabbit spinal ganglia by stereological methods using the electron microscope. Both neurons and their associated satellite cells were examined. In neurons, the shape and structure of lipofuscin bodies differed in young adults (12-months-old) compared to aged rabbits (79-months-old), whereas substantial changes were not observed in satellite cells. Both in nerve and satellite cells, lipofuscin bodies were scattered singly in young adults, but were often clustered in old animals. Lipofuscin occupied an average of 0.36% of neuronal perikaryal volume in the young adults and 2.55% in the aged rabbits; these percentages are much lower than those observed in the same neurons of other species. In the satellite cells, the corresponding values were 0.29% and 2.02%. In the young adults, the mean size of lipofuscin bodies was significantly greater in neurons than in satellite cells; the size of these bodies increased with age by about 2.7 times in neurons and by about 1.7 times in satellite cells. Consequently, in the old rabbits the mean size of lipofuscin bodies was about 2.2 times greater in neurons than in satellite cells. It has been suggested that lipofuscin bodies are transferred from the neuronal perikaryon to the surrounding satellite cells and then removed via the capillaries. However, the present findings suggest that lipofuscin located in satellite cells represents pigment formed by these cells rather than being transferred from neuronal perikaryon. It would appear that the age-related accumulation of lipofuscin in the rabbit spinal ganglia has little effect on neuronal metabolism.

Aging↗

Age-related decrease in the overall extent of perikaryal projections in rabbit spinal ganglion neurons.

The overall extent of the perikaryal projections of sensory neurons from spinal ganglia of young adult and aged rabbits was estimated by a stereological method using the electron microscope. The extent of perikaryal projections was significantly smaller in the aged animals. This age-related decrease did not seem to depend on factors intrinsic to the neuron, but on the absence of a satellite cell covering over extensive portions of the nerve cell body surface. This decrease may influence the organization of the subplasmalemmal cytoskeleton, metabolic exchange between the nerve cell body and its environment and perhaps also neuronal metabolism.

Aging↗

Age-related decrease of the perineuronal satellite cell number in the rabbit spinal ganglia.

This study was undertaken to establish whether a change in the perineuronal satellite cell number contributes to the age-related reduction of the volume ratio between the perineuronal glial sheaths and their associated nerve cell bodies, observed to occur in rabbit spinal ganglia. The volumes of the nerve cell bodies and the numbers of the related satellite cell nuclei were estimated on serial semithin sections from young adult and old rabbits. As satellite cells are mononucleate, the number of the nuclei corresponds to that of these cells. The satellite cell sheaths in both age groups were also examined under the electron microscope. The mean number of satellite cells was significantly smaller in the aged animals than in the young adults although the mean volume of the nerve cell bodies was significantly larger in the former. Cytoplasmic vacuoles, invaginations of the connective tissue and autophagic vacuoles were more frequent in the old rabbits. Satellite cells with pyknotic nuclei and remnants of degenerated satellite cells were only found in aged animals, although rather rarely. The decrease in the satellite cell number is one of the mechanisms by which the age-related reduction of the volume ratio between the perineuronal glial sheaths and their associated nerve cell bodies takes place. The decrease in the satellite cell number seems to occur, at least in part, through cell degeneration. However, other mechanisms (e.g., detachment of satellite cells from the perineuronal sheaths) cannot be excluded. Since satellite cells play a role in neuronal support, the significant decrease in their number probably has negative consequences for neuronal activity.

Aging↗

Cell body volume of spinal ganglion neurons: estimation by three different methods.

We estimated the mean volumes of two series of nerve cell bodies, one from rabbit and one from rat spinal ganglia by three different methods: a procedure we devised 25 years ago (the circle-fitting method), one of the new stereological methods (the nucleator method) and the method of serial sectioning--the most direct and accurate procedure presently available for estimating cell size. In the case of the rabbit, in which most spinal ganglion neurons have a single nucleolus, the mean volumes estimated by the first two methods are closely similar and deviate by less than 2% from the mean obtained by serial sectioning. In the case of the rat, in which approximately half of the spinal ganglion neurons have more than one nucleolus, the mean volumes estimated by the first two methods are again closely similar, but deviate by about 12% from the mean obtained by serial sectioning. These findings show that: a) both the nucleator method and the circle-fitting procedure are more accurate when applied to neurons with a single nucleolus; b) if certain conditions are respected, not all the methods previously used to estimate cell size give biased results. However, the new stereological procedures are easier and quicker to use than the earlier methods. These findings also show that our previous results obtained by the circle-fitting method are to be considered valid.

Animals↗

Ultrastructural localization of actin in the cell body of rat spinal ganglion neurons.

We used phalloidin staining and immunocytochemistry at the light and electron microscope level to determine the localization of actin in the cell bodies of rat spinal ganglion neurons. The results show that actin is mostly concentrated along the periphery of the neuronal perikaryon, including the perikaryal projections. This localization places actin in a strategic position to be influenced by incoming signals and to produce mechanical tensions able to shape the perikaryal surface.

Actins↗

Age-related reduction of the satellite cell sheath around spinal ganglion neurons in the rabbit.

The volumes of the nerve cell bodies and those of the enveloping satellite cell sheaths from spinal ganglia of young adult and aged rabbits were determined by morphometric methods using the electron microscope. The mean volume of the nerve cell bodies was greater in the old rabbits than in young adults; this is probably related to the larger body size of the old animals. The mean volume of the satellite cell sheaths was, however, smaller in the aged rabbits than in the young adults. Consequently the volume ratio between the satellite cell sheaths and the related nerve cell bodies was significantly smaller in the aged animals. Since satellite cells play an important role in the support of the neuron, the reduction in volume of the perineuronal sheath could be associated with a decrease in the trophic activity of satellite cells towards the enveloped neuron with consequences for neuronal activity. Furthermore, in the satellite cell sheaths of old rabbits, the number and extension of gaps that leave the neuronal surface directly exposed to the basal lamina were significantly increased. Since spinal ganglia lack a blood-nervous tissue barrier, only the satellite cell sheath controls the traffic of material to the nerve cell body. Because the neuronal surface unprotected by the satellite cell envelopment is significantly more extensive in the spinal ganglia of old rabbits than in those of young adults, the nerve cells of the former are more exposed to potential damage by harmful substances. A dense undercoating was seen very frequently beneath the portions of the neuronal plasma membrane not covered by satellite cells.

Aging↗

The black reaction.

Camillo Golgi (1843-1926) invented the black reaction in 1873, when he was head physician at the hospice for old people in Abbiategrasso, near Milan. Unlike the procedures that were available before its invention, the black reaction was able to reveal neurons in their entirety, i.e., with all their processes. This weighty event at first passed unnoticed. The first stirring of interest in the black reaction outside Italy began in 1885. The reasons the Golgi technique took so long to receive wide international attention are here analyzed. After it became known, the black reaction was widely employed for almost 30 years, during which time it was responsible for bringing about major advances in our knowledge of the microscopic anatomy of the nervous system, as well as in other fields of study. A number of results obtained by other researchers with the black reaction were vitally important for establishing the neuron theory. In the period between the two World Wars, the Golgi technique was almost forgotten, but returned in vogue once more around the middle of the 20th century following the introduction of the electron microscope to neurocytological research. One-hundred and twenty years after its invention, the black reaction is still widely employed, not only in combination with electron microscopy, but also as an autonomous technique for light microscope studies on the organization of the nervous system in normal conditions and after experimental manipulations.

Humans↗

On the influence of the perineuronal microenvironment on the outgrowth of perikaryal projections of spinal ganglion neurons.

While the outgrowth of the slender projections from the perikaryon of spinal ganglion neurons is an intrinsic property of these neurons, it is also influenced by the surrounding microenvironment. To obtain evidence concerning whether the outgrowth of these projections is influenced by one or both components of the perineuronal microenvironment (satellite cells plus extracellular matrix) we have taken advantage of a rare arrangement of these neurons. In the spinal ganglia of adult animals nerve cell bodies are occasionally arranged in pairs, the two nerve cell bodies of the pair being separated by a satellite cell sheet lacking a basal lamina, while along the remaining portions of their surfaces they are enveloped by a satellite cell sheath, in turn surrounded by a basal lamina and connective tissue. By studying these paired nerve cells we have been able to compare, in the same nerve cell body, the extent of the perikaryal projections in surface domains associated only with satellite cells and in surface domains associated with both satellite cells and extracellular matrix. In spinal ganglia of the rat and lizard we have found that the overall development of the perikaryal projections does not differ significantly in either of these surface domains. This finding suggests that neuron-satellite cell interactions rather than factors in the extracellular matrix play a role in promoting the outgrowth of perikaryal projections from spinal ganglion neurons.

Animals↗

Perikaryal projections of spinal ganglion neurons: quantitative differences between membrane domains in contact with different microenvironments.

The perikarya of spinal ganglion neurons display numerous slender projections. In the present investigation we have studied whether the extent of these projections is uniform over the entire perikaryal surface or whether there is a difference between the regions of the perikaryon in contact with different microenvironments. In spinal ganglia of the rat and the lizard we have analysed about 200 neuronal cell bodies arranged in pairs and have compared the extent of the projections quantitatively in the areas of interneuronal contact with that in the areas of neuron-to-satellite cell contact. In both species we have found that the projections are present over the entire perikaryal surface and that the overall development of the perikaryal projections is significantly greater in those portions of the surface in contact with satellite cells than in the portions in contact with another neuron. On the basis of these observations we conclude that the outgrowth of perikaryal projections is an intrinsic property of the nerve cell body which is manifested over the entire perikaryal surface; there is, however, an extrinsic influence from the microenvironment of the neuron, which may account for the quantitative differences in different domains of the perikaryal surface.

Animals↗

The percentage of nerve cell bodies arranged in clusters decreases with age in the spinal ganglia of adult rabbits.

In the spinal ganglia of the rabbit the nerve cell bodies, which in early developmental stages are mutually in contact, come to be completely isolated from each other by a satellite cell sheath and by a connective envelope before birth. The present study demonstrates that in the early postnatal months some nerve cell bodies are still arranged in clusters, and that the percentage of these decreases progressively throughout adult life. This decrease probably arises because in some of the ganglion neurons the process of envelopment of the perikaryon by an individual sheath begins later, or takes place more slowly, than in the majority of cases. Therefore, the relationship between neurons and between neurons and satellite cells may change in certain clusters of nerve cell bodies under normal circumstances during adult life.

Aging↗