Search PubMed⌕ Search

Biomedical subjects

G Gabella

Publications and source records attributed to G Gabella.

At least 55 records · Page 3Linked to original sources

Tracheal parasympathetic neurons of rat, mouse and guinea pig: partial expression of noradrenergic phenotype and lack of innervation from noradrenergic nerve fibres.

Noradrenergic nerves were studied in whole-mount preparations of the rat, mouse and guinea pig trachea by means of glyoxylic acid-induced catecholamine fluorescence and dopamine beta-hydroxylase immunoreactivity. In an effort to raise tissue levels of catecholamines, some specimens were also treated with the monoamine oxidase inhibitor pargyline, and with L-DOPA, a precursor of noradrenaline. Noradrenergic nerve fibres were detected around blood vessels, within the tracheal smooth muscle and in the mucosa, but never around or in the proximity of neurons of the tracheal ganglia, even after amine precursor loading. These parasympathetic ganglion cells did not show catecholamine fluorescence under control conditions. In the rat and mouse, but not in the guinea pig, some tracheal neurones were dopamine beta-hydroxylase immunoreactive and showed uptake and metabolism of amine precursors, thus expressing aspects of the catecholaminergic phenotype.

Adrenergic Fibers↗

Innervation of the guinea pig trachea: a quantitative morphological study of intrinsic neurons and extrinsic nerves.

The innervation of the guinea pig trachea was studied in wholemount preparations stained for acetylcholinesterase, catecholamines, and substance P immunoreactivity and by electron microscopy. The majority of parasympathetic and afferent nerve fibres arrive from the vagus via branches of the recurrent laryngeal nerves. The recurrent laryngeal nerves are composed of several fascicles comprising 600-700 small myelinated fibres (2-5 microns diameter) and about 1,000-2,000 unmyelinated fibres; both components exit from the nerve and project in fine branches to the trachea. A separate component of 200-250 large myelinated fibres (more than 5 microns diameter) runs the full length of the nerve and innervates the striated muscles of the larynx. The recurrent laryngeal nerves are slightly asymmetric in their origin, length, number, and composition of fibres, with the right nerve being shorter but with more numerous and thinner myelinated fibres. At the distal end of the recurrent nerve, a fine branch called the ramus anastomoticus connects it to the superior laryngeal nerve. In the tracheal plexus, there are on average 222 ganglion cells (range 166-327), distributed mostly in small ganglia of 12 or fewer neurons. The ganglionated plexus is situated entirely outside the tracheal wall, overlying the smooth muscle. Ligation experiments show that sympathetic nerve fibres reach the trachea with the recurrent nerves via anastomoses between the sympathetic chain and vagus nerves, or occasionally with recurrent nerves directly, the largest being at the level of the ansa subclavia. There are also perivascular sympathetic nerve plexuses. Substance P immunoreactive fibres enter the trachea from the vagus nerves and by pathways similar to those of sympathetic nerves. There are also paraganglion cells within the recurrent laryngeal nerve that contain catecholamines and are surrounded by substance P immunoreactive fibres. After cervical vagotomy, all the large myelinated fibres of the ipsilateral recurrent laryngeal nerve degenerate and so do all but 10 or 20 small myelinated fibres and all but a few unmyelinated fibres. Degenerating fibres are found within the entire tracheal plexus, indicating bilateral innervation. The small myelinated fibres that survive cervical vagotomy probably represent sympathetic or afferent nerves with their cell bodies located in sympathetic or dorsal root ganglia.

Acetylcholinesterase↗

Development of smooth muscle: ultrastructural study of the chick embryo gizzard.

The growth and differentiation of smooth muscle in the chicken gizzard were studied by electron microscopy from the 10th day in ovo to 6 months after hatching; during this period the organ grows 1000-fold in weight. At the earliest stage studied, smooth muscle cells, interstitial cells, and fibroblasts are immature but can already be clearly distinguished. The structural components of muscle cells develop in a characteristic sequence. Mitochondria are more abundant in immature muscle cells (8% in 14 days embryos and 7% in 19 days embryos) than in the adult (5%). Caveolae are virtually absent in the 11 day embryo; they become more common at the end of embryonic life, but continue to increase in relative frequency after hatching. Gap junctions appear around the 16th day in ovo as minute aggregates of connexons, which then grow in size, probably by addition of new connexons. In the earliest stages studied, myofilaments occupy 25% of the cell profile and are assembled into bundles accompanied by dense bodies and surrounded by loosely arranged intermediate filaments. By contrast, membrane-bound dense bands are scarce until the latter part of embryonic life, an observation suggesting that myofilament formation and alignment is not a process initiated near the cell membrane or directed by the cell membrane, and that only late in development bundles of myofilaments become extensively anchored to dense bands over the entire cell surface: at that time myofilaments occupy more than 75% of the cell volume. The muscle cells increase about four-fold in volume over the period studied; the 1000-fold increase in muscle volume is mainly accounted for by an increase in muscle cell number. Mitoses are found in the gizzard musculature at all embryonic ages with a peak at 17-19 days; they occur in muscle cells with a high degree of differentiation. These cells divide at a stage when they are packed with myofilaments and form junctions with neighbouring cells: the mitotic process affects the middle portion of the cell, which takes up an ovoid shape and eventually divides, whereas the remaining portions of the cell do not differ in appearance from the surrounding muscle cells. At all stages of development the population of muscle cells has a uniform appearance (apart from the cells in mitosis), and the growth and differentiation seem to proceed at the same pace in all the cells. There are no undifferentiated cells left behind in the tissue for later development.

Animals↗

Fall in the number of myenteric neurons in aging guinea pigs.

The neurons of the myenteric plexus of the entire small intestine were stained in young adult (3-4 mo old) and aging guinea pigs (26-30 mo old). Total length and circumference of the intestine were measured in the same experiments. The small intestine of the aging guinea pigs was longer, and (in the conditions of distention used) had a total serosal surface approximately 70% greater than in young adult animals. The spatial density of myenteric neurons per unit of serosal surface fell dramatically in aging animals, and the total number of myenteric neurons in the small intestine ranged between 1.1 and 1.6 million, i.e., it was only 40%-60% of the value obtained in young adult guinea pigs (2.75 million). The light microscope appearance of the neurons of the two groups of animals was markedly different and the suggestion is put forward that in aging guinea pigs the substantial reduction in neuron number is accompanied by structural changes and reorganization of the neurons that are left.

Aging↗

The intestine as a model for neuronal plasticity.

This study focuses on establishing the nature and extent of the changes that occur in gastroenteric innervation, specifically the myenteric plexus of the rat ileum, following an injury generated by experimental obstruction. A partial obstruction was accomplished by placing a cuff around the terminal portion of the ileum of the rat. Substantial hypertrophy of the enteric muscle wall occurred after 3-5 weeks. Light microscopic examination of the myenteric plexus revealed changes in the numbers of neuronal perikarya, ganglia and perikarya per ganglia; sizes and shapes of perikarya; and thicknesses of nerve fiber bundles. Using vasoactive intestinal polypeptide (VIP) and substance P light microscopic immunohistochemistry, we observed indications of transmitter accumulation in cell bodies and nerve fibers and reactive, degenerative and regenerative changes in axonal endings. Electron microscopic studies provided evidence for neuroplastic changes, as demonstrated by the appearance of reactive and regenerative, or growth, cones in the myenteric plexus.

Animals↗

Structure of the guinea-pig trachea at rest and in contraction.

The trachea of the guinea-pig measures about 47.5 mm in situ, and it shrinks to 38 mm when excised. It can be stretched to the in situ length with a load of 2-4 grams. The transverse area of its lumen measures about 4.5 mm2 in the cervical portion, whereas in the lowermost thoracic portion it measures 2.8 mm2, a difference of 37%. The lumen has an oval shape with the transverse diameter always exceeding the sagittal diameter. The separation between the ends of a cartilage in the dorsal region of the trachea is greater in the cervical than in the thoracic region. Elastic fibres are abundant in the tunica propria and run longitudinally; the collagen is in some areas arranged with a criss-cross pattern. The muscle is inserted on the concave aspect of each cartilage at some distance from the ends of the cartilage and it is therefore much longer than the gap between cartilage ends. Upon contraction in vitro (induced by carbachol) the muscle shortens by about 50%; there is a marked decrease of the transverse diameter of the trachea, and a certain decrease also of the sagittal diameter due to a straightening of the muscle and a change in shape and a movement of the mucosa. The cartilage ends are brought together and in the thoracic region they are bent and overlap extensively. The lumen of the trachea becomes circular and its area is reduced to 2.2 mm2 in the cervical portion and 1.7 mm2 in the thoracic portion.

Animals↗

Quantitative cytology of ganglion neurons and satellite glial cells in the superior cervical ganglion of the sheep. Relationship with ganglion neuron size.

Neurons and glial cells of the superior cervical ganglion of sheep were investigated with morphometric methods in the light and electron microscope. The nerve cell sectional area (measured on nucleated cell profiles) ranged from 165 to 2500 microns2, which corresponds to range in cell diameter from 14 to 56 microns and a range in cell volume from 1600 to 93,000 microns3, i.e. a 60-fold volume difference between smallest and largest neurons. The distribution of cell sizes appeared unimodal, with a predominance of small neurons; there were no variations in different parts of the ganglion. This wide range in nerve cell sizes is discussed in the light of the suggestion that large neurons innervate a greater amount of target tissue (e.g. smooth muscle) and are less readily excitable than smaller neurons: it is thus possible that there is differential recruitment of ganglion neurons in autonomic reflexes. The ultrastructural features of ganglion neurons in the sheep were similar to those observed in small laboratory animals. The relative volumes of perikaryal cytoplasm occupied by mitochondria and Golgi apparatus were 8.5% and 4.8%, respectively, but the average values were the same in small and large neurons. Subsurface cisternae of endoplasmic reticulum were common in the perikaryon, while in the dendrites clusters of synaptic vesicles were found beneath the plasma membrane; the absence of a glial wrapping at the latter sites suggests that they are points of (non-synaptic) release of transmitters. The extent of the capsule that satellite cells form around each neurons was compared in size-based classes of neurons. There was no difference in the size of glial nuclei, and this suggests that glial cells are probably of uniform size. However, glial cells were more densely packed over the surface of large neurons than over the surface of small neurons--in fact the packing density was proportional to the ganglion neuron volume, rather than to its surface. The average thickness of the glial capsule was significantly greater around large than around small neurons. It is suggested that the matching of glial cell number and nerve cell volume is achieved during development by glial cell mitosis taking place long after the nerve cells have ceased dividing.

Animals↗

Neurogenic inflammation in the rat trachea. II. Identity and distribution of nerves mediating the increase in vascular permeability.

This study addresses the question of whether increased vascular permeability, which is a prominent feature of neurogenic inflammation in the respiratory tract, is mediated by sensory axons that end near venules in the airway mucosa. In these experiments, neurogenic inflammation was produced in the tracheal and bronchial mucosa of atropine-treated Long-Evans rats by electrical stimulation of the left or right superior laryngeal nerve and/or cervical vagus nerve. The particulate tracer Monastral blue was injected intravenously to localize the sites of increased vascular permeability, and microspectrophotometry was used to measure the amount of extravasated Monastral blue in the trachea and thereby quantify the increase in vascular permeability. In some rats, selective denervations were made to locate the cell bodies of neurons that mediate the increase in vascular permeability; in others, fluorescence immunohistochemistry and quantitative electron microscopic methods were used to determine which structures in the tracheal mucosa are innervated by these neurons. The study revealed that the vagally mediated increase in vascular permeability was sudden, transient (half-life = 2.4 min) and restricted to venules. Stimulation of the left or right superior laryngeal nerve increased the permeability of venules in the extrathoracic trachea, whereas stimulation of either vagus nerve increased vascular permeability in the intrathoracic trachea and bronchi. All nerves had bilateral effects in the trachea, but the vagus nerves had largely unilateral effects in the bronchi. Neurons that mediated the increase in venular permeability had their cells bodies in the jugular (superior sensory) ganglion of the vagus nerve or rostral portion of the nodose (inferior sensory) ganglion. Preganglionic autonomic vagal neurons in the brain stem were not essential for this increase in venular permeability. Few nerves identifiable by substance P-immunohistochemistry or electron microscopy were located near the affected venules, and no nerves were within 1 micron of the walls of venules. However, the epithelium and arterioles of the airway mucosa were densely innervated. All intraepithelial nerves were within 0.1 micron of epithelial cells, and at least two-thirds of nerves near arterioles were within 1 micron of the vessel walls. We conclude that the increase in venular permeability associated with neurogenic inflammation in the trachea and bronchi of rats is mediated by sensory axons that travel in the vagus nerves and superior laryngeal nerves.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Ureteric ganglia in the guinea pig.

We studied the ureter of guinea pigs to ascertain whether there are ganglia and ganglion neurons directly associated with this structure. We consistently observed ureteric ganglia and individual neurons in the adventitia of the ureter or very close to it. There were on average 87 neurons per ureter, and they were scattered over the entire length of the tube.

Animals↗

The cross-ply arrangement of collagen fibres in the submucosa of the mammalian small intestine.

Whole-mount preparations of the submucosa were made from the small intestine of rats, guinea-pigs, rabbits and sheep. In the distended intestine the collagen fibres ran straight and approximately parallel to the serosal surface. They formed a characteristic lattice, with two arrays of fibres running diagonally in a clockwise and an anticlockwise direction, and making an angle of 50 degrees-55 degrees with the longitudinal axis of the intestine. This collagen-fibre lattice was flexible and changed with the movements of the intestinal wall; when the radial distension predominated, the angle between collagen fibres of the submucosa and longitudinal axis of the intestine increased to 60 degrees-65 degrees, and when the longitudinal distension predominated the angle decreased to about 30 degrees.

Animals↗

Scanning electron microscopy of the muscle coat of the guinea-pig small intestine.

We have studied the layers of the muscular coat of the guinea-pig small intestine after enzymatic and chemical removal of extracellular connective tissue. The cells of the longitudinal muscle layer are wider, have rougher surfaces, more finger-like processes and more complex terminations, but fewer intercellular junctions than cells in the circular muscle layer. A special layer of wide, flat cells with a dense innervation exists at the inner margin of the circular muscle layer, facing the submucosa. The ganglia of the myenteric and submucosal plexuses are covered by a smooth basal lamina, a delicate feltwork of collagen fibrils, and innumerable connective tissue cells. The neuronal and glial cell processes at the surface of ganglia form an interlocking mosaic, which is loosely packed in newborn and young animals, but becomes tightly packed in adults. The arrangement of glial cells becomes progressively looser along finer nerve bundles. Single varicose nerve fibres are rarely exposed, but multiaxonal bundles are common. Fibroblast-like cells of characteristic shape and orientation are found in the serosa; around nerve ganglia; in the intermuscular connective tissue layer and in the circular muscle, where they bridge nerve bundles and muscle cells; at the submucosal face of the special, flattened inner circular muscle layer; and in the submucosa. Some of these fibroblast-like cells correspond to interstitial cells of Cajal. Other structures readily visualized by scanning electron microscopy are blood and lymphatic vessels and their periendothelial cells. The relationship of cellular elements to connective tissue was studied with three different preparative procedures: (1) freeze-cracked specimens of intact, undigested intestine; (2) 'stretch preparations' of longitudinal muscle with adhering myenteric plexus; (3) sheets of submucosal collagen bundles from which all cellular elements had been removed by prolonged detergent extraction.

Animals↗

On the nerve plexus of the chicken gizzard.

The Auerbach's plexus of the gizzard was stained in toto in adult chicken and in young and newly-hatched chicks. The plexus lies immediately beneath the serosa and extends over 55% of the surface of the organ, covering its cranial and caudal poles and the two curvatures. The areas into which the plexus does not extend (i.e., most of the ventral and the dorsal surface) are those where the muscle is covered by the laminar tendon of the gizzard. The ganglia are large, often with hundred of neurons, and have short and broad connecting strands. They are surrounded by a capsule of connective tissue. The ganglion neurons are discoidal and in the adult they measure up to 50 microns in diameter, each being surrounded by a set of glial cells. A few small neurons persist in the adult; in the newly-hatched chick these are predominant, but some large neurons up to 25 microns in diameter are already present. The ultra-structural features of the ganglia of the Auerbach's plexus include the abundance of axo-somatic synapses, as well as numerous axo-dendritic synapses, the presence of intra-ganglionic bundles of collagen fibrils and blood vessels, the abundance of glial cells. In addition to the plexus beneath the serosa, the gizzard has many small intramuscular ganglia located throughout the musculature (which is exclusively circular). These ganglia do not have a connective tissue capsule and are made of small and tightly packed neurons.

Animals↗

Fine structure of the autonomic ganglia of the mouse pulmonary vein.

The aim of this study was to describe the architecture of a ganglionated nerve plexus found in the loose connective tissue surrounding the pulmonary vein of the mouse. The input to this plexus was from the vagus nerves and from the stellate ganglia. A large ganglion containing more than 200 neurons was commonly found near the primary bifurcation of the pulmonary vein. The neurons were studied by NADH-diaphorase, zinc iodide-osmium and glyoxylic acid-induced catecholamine fluorescence methods at the light microscopic level, by scanning electron microscopy after the removal of connective tissue, and by transmission electron microscopy. The shape of the neuronal cell bodies was generally a smooth ellipsoid with the average major axis about twice the minor axis. The measured maximum cell diameter ranged from 14 to 42 micron (mean 26 micron). The profile area of individual neurons, as measured from wholemount preparations, ranged from 100 to 800 micron2 (mean 340 micron2) and the calculated neuronal volume ranged from 500 to 12,000 micron3 (mean 3300 micron3). Although there was this wide spread in neuronal size, histograms of cell size showed no separate populations of neurons. Almost all of the ganglionic neuronal cell bodies showed no catecholamine-specific fluorescence, but about 1% of the neurons exhibited a weak green fluorescence. Only a few noradrenergic nerve fibres were seen within the ganglia and these were associated with intraganglionic blood vessels. Small, intensely fluorescent cells were only rarely associated with the ganglia. Neurons and satellite cells formed units which were surrounded by an intraganglionic connective tissue space and a perineurium. Some of the intraganglionic capillaries were fenestrated. Neurons were entirely surrounded by satellite cells and did not appear to have any long dendrites. The generally smooth neuronal cell bodies had short spine-like processes, which were confined to within the satellite cell sheath. Preganglionic nerve fibres formed pericellular baskets of varicose fibres around neurons and made synapses either directly on the cell body or on somatic spines in about equal numbers. No synapses were found in the neuropil at a distance from the neuronal cell body. A few nerve processes were deeply embedded within the neuronal cell body. Clusters of vesicles were found in the cytoplasm of most neurons and were associated with subplasmalemmal densities. These synapse-like structures were mostly directed towards satellite cells, but some were associated with incoming synapses.

Animals↗

The number of neurons in the small intestine of mice, guinea-pigs and sheep.

A histochemical technique was used to stain the myenteric neurons in the intact wall of the small intestine of mice, guinea-pigs and sheep. The length and diameter of the small intestine and the total serosal surface area were also obtained. Myenteric neurons were counted on large whole-mount preparations of the muscularis externa. Counts were carried out also on the submucosal plexus, on a more limited scale. In the mouse a spatial density of 10,600 myenteric neurons per cm2 was found. The small intestine was 33 cm long and measured on average 11.5 mm in circumference, the total outer surface (serosal surface) amounting to about 38.0 cm2. The total number of myenteric neurons in the small intestine was calculated as about 403,000. In the guinea-pig the length of the small intestine was 145 cm, the average circumference 22 mm and the total outer surface area about 319 cm2. The neuronal packing density was 8600/cm2, and the total number of myenteric neurons about 2,750,000. In the sheep the small intestine was about 2100 cm long with an average circumference of 60 mm and a total surface area of about 12,600 cm2. The ganglion neuron density was about 2500/cm2, and the total number of myenteric neurons in the small intestine was calculated as about 31,500,000. Thus, in the sheep the small intestine contained about 11 times as many myenteric neurons as the guinea-pig and about 80 times as many as the mouse. The differences are in the same direction as, but not proportional to, the differences in body weight and in the length of the intestine. The neuronal spatial density was highest in the mouse and lowest in the sheep, and in the sheep the neurons were markedly larger, and gathered in ganglia that were larger and further apart from one another, than in the mouse, while they had intermediate values in the guinea-pig. A new way of expressing neuronal packing densities is presented together with the proposal of an arbitrary but reproducible unit of intestinal length (a segment whose length is equal to its diameter). In the submucosal plexus the neuronal density was about 3000/cm2 in the guinea-pig, about 8700/cm2 in the mouse and about 4500/cm2 in the sheep. In the mouse the submucosal neuron density decreased gradually along the length of the small intestine.

Animals↗

Quantitative study of the ganglion neurons of the mouse trachea.

The ganglionated plexus of the trachea of mice was studied quantitatively with a histochemical method that stains electively the ganglion nerve cells in whole-mount preparations. The plexus lies exclusively over the muscular part of the trachea, dorsal to the muscle itself, and it varies considerably in pattern and extent between individual animals. In young adult mice the plexus contains on average 235 neurons, occurring singly or gathered in packed ganglia. The ganglion neurons are relatively small, the profile area of three quarters of them measuring between 150 and 275 microns 2 with an average of 251 microns 2. In ageing mice the average number of ganglion neurons is the same as in young animals; however, cell sizes are markedly increased, the average being 341 microns 2. Among the ultrastructural features of the ganglia, is a capsule (perineurium) of very regular structure, the presence of collagen, capillaries and myelinated axons inside the ganglia, and the presence of only few and short dendrites, some of which are abutted by synapsing nerve endings.

Age Factors↗