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Biomedical subjects

G Gabella

Publications and source records attributed to G Gabella.

At least 19 recordsLinked to original sources
PubMed1998

Glial fibrillary acidic protein (GFAP) immunoreactivity in enteric ganglia of the chick embryo.

We examined by immunohistochemistry the expression of glial fibrillary acidic protein (GFAP) in enteric ganglia of the chick embryo, using a polyclonal antibody. The morphology of enteric ganglion cells was examined by electron microscopy. Faint GFAP immunoreactivity was detected in ganglion cells and cell processes from around day 7 in ovo. Later in development the intensity of the immunofluorescence increased and it became more evident that immunoreactive small ganglion cells (interpreted as primitive glial cells), and their processes, surrounded larger negative cell profiles (interpreted as primitive neuronal cells); GFAP immunofluorescence was also evident in intramuscular and mucosal nerve trunks. In colocalization experiments, GFAP immunoreactivity was detected in a proportion of HNK-1/N-CAM immunoreactive ganglion cells, in both the myenteric and submucosal plexus. In addition, we observed GFAP immunoreactive nerves in wholemount preparations of chick gut from as early as day 4.5 in ovo. In the ganglionated nerve of Remak, GFAP immunoreactive satellite and Schwann cells were in evidence from day 5 of incubation. Neuronal markers, such as neurofilament, have been detected very early in development in neural crest cell populations in chick enteric ganglia. In contrast, the expression of markers of the glial phenotype has previously been observed only in the late stages of embryonic development. From our experiments, we conclude that neuronal and glial phenotypes are immunohistochemically distinct from as early as day 4.5 of incubation, even if by ultrastructural criteria glial cells are clearly distinguishable from neurons only after day 16 in ovo.

Animals↗
PubMed1998

The distribution of intramural nerves in urinary bladder after partial denervation in the female rat.

We evaluated the degree of neuronal plasticity following a partial denervation of the rat urinary bladder. Using acetylcholinesterase staining we found that the postganglionic nerves from the pelvic ganglion reach the intact bladder as 1-4 nerve trunks on each side, slightly ventral and caudal to the ureteral orifices. Normally a few thinner nerves also reach the bladder posterolateral to the ureterovesical junction. The nerves ventral to the ureters run in the ventral longitudinal muscle layer as well-defined trunks with a pattern that does not differ much from one animal to another. The nerves reaching the bladder dorsolaterally innervate the dorsolateral aspects in a more irregular fashion. Some anastomoses are found across the midline between nerves from either side. This nerve pattern is already in place in newborn rats. After removal of the pelvic ganglion on one side in the adult rat the ipsilateral ventral nerves rapidly degenerate, whereas some dorsolateral nerves usually survive. Axons from the intact ventral nerves can be seen crossing over to the denervated side in the anastomoses. After 13 weeks the surviving ventral nerves, which normally run at some distance from the ventral midline, now run in the midline with equal amounts of ventral longitudinal muscle on either side, and with their branches evenly distributed to both sides. The same pattern is seen after 27 weeks. Unilateral ganglionectomy in 3-week-old rats leads to the same changes in nerve distribution as in the adult rat. We conclude that there is a high degree of plasticity in the bladder innervation following a partial denervation, and that this plasticity includes the distribution of its main intramural nerve trunks.

Acetylcholinesterase↗
PubMed1998

Homotransplant of pelvic ganglion into bladder wall in adult rats.

In these experiments a large portion of the pelvic ganglion of adult female rats was transplanted into the wall of the urinary bladder of the same animals. The morphology and fine structure of the transplants were studied in whole-mounts and in sections for light and electron microscopy, from two days up to four months after operation. The general architecture of the ganglion was preserved in all the transplants. The vascularization was re-established. Nerves grew out of the transplant and connections with the original intramural nerves of the bladder wall were established. All the synapses degenerated at the time of transplantation; new synapses began to reappear on the ganglion neurons in the oldest transplants. Although some neurons in the transplant degenerated during the first few days, the majority of neurons survived for the full length of the experiments (four months). Satellite glial cells and small intensely fluorescent cells had a similar structure and distribution as in control ganglia. The results show that the homotransplant of pelvic neurons into the bladder has a high rate of success, in terms of survival, maintenance of fine structure, growth and re-connections; these neurons of adult organisms display plastic and regenerative abilities.

Acetylcholinesterase↗
PubMed1998

Connection of smooth muscle cells to elastic lamellae in aorta of spontaneously hypertensive rats.

We have recently demonstrated that in large arteries of spontaneously hypertensive rats (SHR), there is no increase of stiffness despite the increase in wall thickness, a sign of mechanical adaptation of the arterial wall to the higher level of stress. Because the dense plaques of smooth muscle are a major site of anchorage between the muscle cells and extracellular matrix, we determined by electron microscopy the distribution of dense plaques and their connections to elastic lamellae in the abdominal aorta of 1-year-old SHR and control Wistar rats. In vivo echo-tracking measurement of aortic distensibility and elastic modulus indicates a reduction of arterial stiffness in SHR compared with Wistar rats when they are studied over a common range of blood pressure. The media thickness to body weight ratio was higher in SHR than in Wistar rats. In the media, the percentage of sectional area occupied by extracellular matrix was not different between Wistar rats and SHR. The average number of dense plaques per muscle cell was not different between Wistar rats and SHR. However, the percentage of cell surface occupied by dense plaques was increased in SHR, and the percentage of cell surface connected to the elastic lamellae was twice as high in SHR compared with Wistar rats (9.4+/-1.5% versus 3.8+/-1.1%). These results suggest that the elastin network plays a major role in the mechanical adaptation of the arterial wall in SHR, not through variations of its total amount but through variations of the extent of anchorage to the muscle cells.

Adaptation, Physiological↗
PubMed1997

Laminin immunoreactivity in enteric ganglia of the chick embryo.

The localization and time of appearance of laminin in the duodenum of the chick embryo were studied with an anti-laminin polyclonal antibody and immunofluorescence. Laminin immunoreactivity was observed in the basement membranes of the mesothelium, mucosal epithelium, muscle cells and in the adventitia and basal surface of the endothelium in blood vessels. In addition, laminin immunostaining was detected over the contour of myenteric ganglia from embryonic day 7 and inside these ganglia from embryonic day 13. In colocalization experiments, laminin immunoreactivity occurred outside tubulin immunoreactive neuronal cell bodies, thus indicating that it resides in glial cells or in extracellular spaces. In addition connecting strands of the myenteric plexus and intramuscular nerves expressed laminin immunoreactivity. Similar observations were made in the proventriculus, gizzard, ileum and rectum of chick embryos, and in the duodenum and rectum of quail embryos. In the ganglion of Remak, laminin immunofluorescence was detected in the collagenous sheath that surrounds the ganglion and inside the ganglion, where it outlines neuronal cell bodies. Laminin immunoreactivity within the myenteric ganglia during the 3rd week in ovo, appears to be characteristic of the avian species examined, since it was not observed in the rat and mouse intestine at equivalent developmental stages. Immunocytochemical experiments at the electron-microscope level confirmed that structures with laminin or laminin-like immunoreactivity occur both around and inside myenteric ganglia. It is suggested that laminin, or an immunologically similar molecule, may play a role in the development and maturation of avian enteric ganglia.

Animals↗
PubMed1996

Decrease and disappearance of intramural neurons in the rat bladder during post-natal development.

While confirming previous results that the bladder of adult female rats is devoid of intramural neurons, we show that during postnatal development some intramural neurons are present. There is about 200 of them per bladder at birth, and their number progressively decreases during post-natal life. In this strain of rats some neurons are still present at 12 weeks of age, and in one animal (out of five) there were still 25 neurons at 20 weeks of age.

Animals↗
PubMed1996

Origin of the c-kit-positive interstitial cells in the avian bowel.

Interstitial cells of Cajal (ICC) aroused much interest among neuroanatomists at the beginning of the century. These small cells, organized into networks, are intercalated between nerve fibers and muscle cells, and are now considered by many authors to be responsible for the pacemaker activity of the gut. Renewed interest in these cells arose recently when the receptor tyrosine kinase, c-kit, was shown to be associated with their functional activity. The embryonic origin of interstitial cells has remained a controversial issue ever since their discovery. Some authors consider them to be of neural or glial nature and thus of neural crest origin. Others consider them to be of fibroblastic or muscular nature. We have applied the quail-chick marker system to solve this problem. ICC were identified by means of a chicken-c-kit nucleic probe which cross-reacts with the quail c-kit gene product. We constructed chimeric bowels by grafting isotopically quail vagal neural crest into chick embryos at embryonic day 2 (E2). The enteric innervation of the chimeras was then of quail origin. In situ hybridization of the chimeric bowels showed that all the c-kit-positive cells were of the chick type, and therefore belonged to the gut mesenchyme and were not neural crest-derived cells. This observation was confirmed by culturing aneural chick guts on the chorio-allantoic membrane. Typical ICC, as defined at the EM level and by their expression of the c-kit receptor, developed in the gut wall in the complete absence of enteric innervation. One can conclude the ICC are of mesodermal origin and develop independently from enteric neurons with which they later establish anatomical and functional relations.

Animals↗
PubMed1995

Intramural neurones appear in the urinary bladder wall following excision of the pelvic ganglion in the rat.

The entire bladder of female rats was stained for acetylcholinesterase activity, in order to make visible all the intramural nerves. Ganglion neurones were never observed within the bladder wall of adult controls. In contrast, 2, 13 or 27 weeks after unilateral pelvic ganglion destruction a few intramural neurones were consistently observed along the remnants of nerves in the originally denervated half of the bladder. These neurones were often gathered into clusters of 5-15, inside a nerve or closely connected to it, with a faintly stained nerve leading to them and a more heavily stained nerve leading from them. The origin of the new intramural ganglion neurones is unknown, but they probably migrate after ganglionectomy, possibly from some accessory ganglion close to the bladder.

Acetylcholinesterase↗
PubMed1995

Complex structure of the common carotid artery of sheep.

BACKGROUND: Mechanical properties of blood vessels are dictated by the vessel wall structure. In many large conduit vessels the tunica media is a sheath of circular musculature and the tunica adventitia a layer of fibrous connective tissue with limited longitudinal extensibility. In contrast, the carotid artery of the sheep displays in each tunica a more complex architecture of muscle and connective tissue. METHODS: Vessels collected from ewes were measured and processed for light microscopy and for transmission electron microscopy. RESULTS: Layers of histologically different materials are found within the tunica intima, media and adventitia. (1) The tunica media is made of circumferentially arranged muscle cells markedly different at different depths. In the innermost third of the media, muscle cells are small and with irregular profiles, the cells are widely separated, and the extracellular material is abundant and composed mainly of elastic fibres. In the outermost third, muscle cells are larger and with more regular profiles, the cells are relatively close to each other and the extracellular material is sparse and consists mainly of collagen fibrils. (2) A small number of fibroblasts is found in all parts of the media amongst the preponderant muscle cells. (3) The intima contains fibroblast-like cells and longitudinally arranged muscle cells. (4) The adventitia contains a thick layer of collagen and elastic fibres; external to this, it displays a conspicuous musculature, made of large bundles of longitudinal muscle. CONCLUSIONS: The carotid artery of the sheep presents in all three coats of its wall features which are at variance from those in the better known vessels of small laboratory animals. The presence of many layers of material within the wall, the heterogeneity of the tissues found, and the occurrence of an extensive longitudinal musculature, have important effects on the mechanical properties of the vessel.

Animals↗
PubMed1995

The structural relations between nerve fibres and muscle cells in the urinary bladder of the rat.

Intramuscular nerve fibres in the bladder of adult female rats were investigated by means of serial sections. The following observations were made. (1) Upon penetrating into the musculature the nerve bundles branch repeatedly, and almost all turn into single fibres; their axons become varicose, the Schwann cell sheath is attenuated, incomplete or absent, and the separation between axonal membrane and muscle cell membrane is reduced to tens of nanometres. (2) All single axons, and some of those within bundles, are varicose, but the characteristic of being varicose is expressed by degrees, and is not an all-or-none state. (3) Varicosities contain vesicles (mostly of the agranular type), microtubules (with little connection with the axolemma or the vesicles), some neurofilaments (scarce or absent in the best developed varicosities), mitochondria (whose size is on average smaller than those of the perikaryon, and a minute amount of endoplasmic reticulum. (4) Terminal varicosities, the true anatomical ending of an axon, are often devoid of Schwann cell sheath, are packed with vesicles, rarely contain microtubules or neurofilaments, and lie close to a muscle cell: the gap is often reduced to approximately 10 nm. (5) Schwann cells accompany the axons within the muscle strands. Unlike the area of the axonal profiles, the area of glial sheath changes little along the length of the nerve fibre, except towards its end. (6) The Schwann cell sheath around a varicosity is often incomplete; the area of the axolemma thus exposed is covered by the basal lamina, and is here referred to as a 'window'. While some varicosities have a window only a few tens of nanometres in width, others have more than one window, and some are devoid of Schwann cell altogether, so that their entire axolemma is in contact with the basal lamina. The Schwann cell never extends beyond the axon, whereas very often (and possibly always) the axon extends beyond the Schwann cell. (7) Intervaricose segments vary in length and diameter, the narrowest ones accompanying the more clear-cut varicosities. Some intervaricose segments are as small as 50 nm in diameter, contain a single microtubule and lack a Schwann cell sheath. Others, sheathed by a Schwann cell, contain a single neurofilament or no organelles at all.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Reversal of muscle hypertrophy in the rat urinary bladder after removal of urethral obstruction.

We studied the ultrastructure of the bladder musculature after first inducing hypertrophy by means of urethral obstruction and subsequently removing the obstruction. With hypertrophy the bladder musculature increases ten-fold or more in volume; after de-obstruction approximately 4/5 of the hypertrophic muscle weight and volume is lost within six weeks. In spite of this very large decrease in muscle mass there is no degeneration of muscle cells or nerve endings or of other cell types in the de-obstructed bladder either at 5 days or at 6 weeks. The individual muscle cells are smaller in size than in the hypertrophic bladder but still larger than control muscle cells. The decrease in muscle cell size is more substantial than the decrease in muscle cell surface. There are no lysosomes or other signs of intracellular degradation in any cells of the muscle layer. The musculature contains a very large amount of intercellular material, mainly collagen. This study documents the great plasticity of the musculature in the reduction of muscle mass after de-obstruction. However, some of the fine structural features are almost as different from the controls as in the hypertrophic muscle.

Animals↗
PubMed1993

Effect of decentralization or contralateral ganglionectomy on obstruction-induced hypertrophy of rat urinary bladder muscle and pelvic ganglion.

After urethral obstruction the musculature of the rat urinary bladder undergoes extensive hypertrophy, irrespective of whether its innervation is intact or whether one of the two pelvic ganglia has been decentralized or excised. Even the excision of both ganglia does not inhibit muscle hypertrophy. The presence of nerves is not a prerequisite for the muscle growth to occur. The stimulus for growth resides in the bladder itself, and the present and previous observations are in agreement with the notion that the distension of the muscle is a primary stimulus for muscle growth. With bladder hypertrophy, the pelvic ganglion neurons undergo hypertrophy, even when they are devoid of their preganglionic input. Synaptic connection with the preganglionic fibres and stimuli from the spinal cord are not prerequisites for neuronal hypertrophy. However, the hypertrophy is less marked in the decentralized ganglion neurons than in the neurons of the contralateral intact ganglion. With bladder hypertrophy and contralateral ganglionectomy, the neuronal hypertrophy is greater than with either procedure alone, suggesting that the two forms of neuronal growth stimulation can be added to each other.

Animals↗
PubMed1993

Effects of unilateral pelvic ganglionectomy on urinary bladder function in the male rat.

Micturition interval and micturition volume were measured in water loaded male rats before and up to 6 weeks after unilateral pelvic ganglionectomy. There was no effect on micturition interval until 7 days after the ganglionectomy. Micturition interval then remained increased. Maximal micturition volume was higher than in the control group from 4 to 14 days after surgery. Cystometrograms were recorded 12 days or 6 weeks after surgery. At 12 days the micturition pressure was lower in the unilaterally ganglionectomized than in the controls. After 6 weeks there was no significant difference in micturition pressure. A reduction of micturition pressure by about 50% was observed after i.v. injection of atropine, but no residual urine developed. Electron microscope investigation showed a considerable decrease in number of nerve terminals on the ganglionectomized side 3 days after surgery. Most of them were devoid of synaptic vesicles. On the contralateral side the majority of nerve terminals appeared normal, but many had a decreased number of vesicles. After 14 days the number of terminals was still lower than normal on the ganglionectomized side. They generally contained only a small number of vesicles. Also, on the contralateral side some nerve endings with the reduced complement of vesicles were found. We suggest that the effects of unilateral ganglionectomy on micturition volume and interval can be explained by a decreased sensory input from the bladder and that the effect on micturition pressure is due to a transient decrease in number and function of motor nerve terminals in the detrusor muscle.

Animals↗
PubMed1992

Hypertrophy and reversal of hypertrophy in rat pelvic ganglion neurons.

An experimental procedure which chronically reduces the lumen of the urethra in adult female rats produced distension of the bladder and conspicuous thickening of its wall, resulting within 6-8 weeks in a ten-fold increase in muscle weight (muscle hypertrophy). During this process, the neurons in the pelvic ganglion that innervate the bladder undergo a large increase in size (neuronal hypertrophy). The average neuronal volume increased by 83%; small neurons became less numerous and large neurons became more numerous than in controls, but there was no increase in the maximum neuronal size. Six weeks after re-operation and removal of the urethral obstruction, the weight of the bladder was reduced (although not quite to the control levels), while the average neuronal size reversed to values very close to controls. In separate experiments, the pelvic ganglion of one side was removed. The nerve fibres in the hemidenervated bladder sprouted, grew and spread to innervate the whole bladder. The neurons in the surviving pelvic ganglion hypertrophied, the average cell volume increasing by 50% in seven weeks. The experiments showed that: (i) the pelvic neurons of adult rats are capable of very extensive growth when the tissue they innervate (bladder muscle) undergoes hypertrophy; (ii) the neuronal hypertrophy is reversible. This was taken to imply that there are factors within the bladder, including trophic substances, that regulate nerve cell volume not only by inducing growth but also by inducing the opposite effect, a cell size reduction; (iii) unilateral ganglionectomy, which did not induce muscle hypertrophy but doubled the amount of muscle innervated by the contralateral ganglion, was followed by marked neuronal hypertrophy.

Animals↗
PubMed1992

Liquid flow across the epithelium of the artificially perfused lung of fetal and postnatal sheep.

1. The lungs of five fetal (133-140 days gestation) and thirty-four postnatal (2-240 days) sheep were artificially perfused in situ with warmed and oxygenated sheep blood. In postnatal animals the airspace of the lung was filled with liquid similar in composition to fetal lung liquid. In fetal and postnatal animals luminal liquid volume was measured by the impermeant tracer technique. 2. Under resting conditions the pulmonary epithelium of fetal animals secreted liquid at a mean (+/- S.E.M.) rate of 2.0 (+/- 0.4) ml (kg body weight)-1 h-1, those of postnantal animals absorbed liquid at -1.8 (+/- 0.2) ml (kg body weight)-1 h-1. 3. Addition of 2,4-dinitrophenol to achieve a concentration of 1.5 x 10(-3) M in the perfusing blood in postnatal animals caused complete cessation of liquid absorption. 4. Light and electron microscopic examination of the lung after periods of up to 6 h of artificial perfusion showed no evidence of epithelial damage. From 3 h onwards, liquid accumulation was evident in the perivascular spaces. 5. Addition of adrenaline to the perfusate in fetal animals caused absorption of liquid to occur at a mean rate of -2.9 (+/- 1.3) ml (kg body weight)-1 h-1. In postnatal animals adrenaline caused the rate of liquid absorption to increase from a mean rate of -1.4 (+/- 0.2) to -2.2 (+/- 0.3) ml (kg body weight)-1 h-1. 6. In the fetus addition of amiloride (0.8 x 10(-4) M) to the luminal fluid blocked adrenaline-induced liquid absorption and caused secretion to occur at 1.3 (+/- 0.3) ml (kg body weight)-1 h-1. 7. In postnatal animals the response to amiloride was age dependent. In newborn lambs (2-14 days) amiloride blocked liquid absorption and caused secretion of liquid to occur in seven out of eight animals at a mean rate of 0.9 (+/- 0.3) ml (kg body weight)-1 h-1 (n = 8). In older animals (15-240 days) the characteristic response to amiloride was slowing of the rate of liquid absorption (mean rate of absorption,-0.2 (+/- 0.09) ml (kg body weight)-1 h-1, n = 18) with liquid secretion being seen in only three of eighteen animals.(ABSTRACT TRUNCATED AT 400 WORDS)

2,4-Dinitrophenol↗
PubMed1992

The interface between epithelium and lamina propria in the rat urinary bladder.

We investigated by transmission and scanning election microscopy the interface between the epithelium and lamina propria in the rat urinary bladder. A digestion technique that dissolves the basal laminae and collagen fibrils was effective in cleaving the mucosa at this level; the specimens were then prepared for scanning electron microscopy, thus visualizing the basal epithelial surface and the uppermost surface of the lamina propria. The underside of the epithelium is scored by very numerous grooves which in the intact organ are occupied by a dense network of blood capillaries. These vascular grooves allow a large number of capillaries (epithelial capillaries) to run at a distance of a few tenths of a micron from the epithelium. On the side of the lamina propria, after collagen and other extracellular materials had been removed, the capillary network itself is visible in the uppermost region. The network is complementary to that of vascular grooves. Other smaller grooves on the basal surface of the epithelium correspond to nerve fibres which run within a few tenths of a micron from the epithelium.

Animals↗
PubMed1991

Ultrastructure of the tracheal muscle in developing, adult and ageing guinea-pigs.

The musculature of the trachea of adult guinea-pigs is ultrastructurally similar to other visceral muscles. However, tracheal muscle cells have irregular outlines, large accumulations of glycogen, a small number of gap junctions, and many small elastic fibres and collagen fibrils (50 nm in diameter). Fibroblasts, mast cells, Schwann cells and axons are found within the muscle, but no interstitial cells. Capillaries run in the connective tissue septa. The tracheal muscle is well differentiated at the end of the fetal life. At this time, muscle cells have an appearance similar to that of a mature muscle. However, the cells are smaller, especially in length, and their orientation is less regular. There are no undifferentiated cells; a few muscle cells are seen in mitosis. There is a limited amount of stroma, mainly small collagen fibrils not exceeding 30 nm in diameter. Within the muscle there are many nerves, but no fibroblasts or mast cells. Many structural contacts link together the muscle cells, but there are no gap junctions. Gap junctions develop in the first few days after birth and are distinct in 5-day-old animals. In ageing guinea-pigs (30-36 months old) the muscle cells are slightly larger than in the young adult. They display deep invaginations of the cell membrane and a very irregular profile. There is an increase in the number of glycogen granule clusters, and the basal lamina is particularly prominent. The amount of stroma has increased, and is made of large elastic fibres and collagen fibrils up to 105 nm in diameter.

Aging↗