Immunohistochemical detection of neuronal plexuses and nerve cells within the upper urinary tract of pigs (original) (raw)
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Immunohistochemical demonstration of nerves and nerve cells in human and porcine ureters
Annals of Anatomy - Anatomischer Anzeiger, 1993
There are divergent opinions about the origin of ureteral motility. It is either a myogenic or a neutral phenomenon. Furthermore, the existence of nerve cells in the ureter is also a controversial question. In the present study we applied immunohistochemical methods to human and pocine ureters in an attempt to clarify the mattem. As neuronal markers we used anti-PGP 9.5 and anti-NSE, and as a glial marker anti-S-100. In the wohle mount preparations of pig ureter we observed two neuronal plexuses on both sides of the tunica muscularis. The inner plexus consisted of both nerve bundles and nerve cells, whereas the outer one did not contain any nerve cells. In the human ureter we found a ganglion with nerve cells beneath the tunica muscularis and the tunica adventitia.
Transitory inner medullary nerve terminals in the cat kidney
Neuroscience Letters, 1990
An indirect immunohistochemical method was used to visualize nerves immunoreactive for tyrosine hydroxylase (THI) and dopamine-fl-hydroxylase (DBHI) in kidney sections of cats 6 weeks and 2 and 3 months of age. THI and DBHI nerve terminals innervate the renal pelvis, interlobar veins and arterial tree including medullary vascular bundles of cats of each age studied. In kidneys of 6-week-old cats, THI and DBHI axons form elaborate plexuses that are distributed throughout much of the inner medulla, whereas some medullary axons appear to degenerate at 2 months and no inner medullary plexuses were visualized in 3-month-old cats. Transitory inner medullary nerves in the cat kidney may influence cellular development and play a role in salt and water balance.
European Journal of Histochemistry, 2013
The urinary bladder trigone (UBT) is a limited area through which the majority of vessels and nerve fibers penetrate into the urinary bladder and where nerve fibers and intramural neurons are more concentrated. We localized the extramural post-ganglionic autonomic neurons supplying the porcine UBT by means of retrograde tracing (Fast Blue, FB). Moreover, we investigated the phenotype of sympathetic trunk ganglia (STG) and caudal mesenteric ganglia (CMG) neurons positive to FB (FB+) by coupling retrograde tracing and double-labeling immunofluorescence methods. A mean number of 1845.1±259.3 FB+ neurons were localized bilaterally in the L1-S3 STG, which appeared as small pericarya (465.6±82.7 µm 2) mainly localized along an edge of the ganglion. A large number (4287.5±1450.6) of small (476.1±103.9 µm 2) FB+ neurons were localized mainly along a border of both CMG. The largest number (4793.3±1990.8) of FB+ neurons was observed in the pelvic plexus (PP), where labeled neurons were often clustered within different microganglia and had smaller soma cross-sectional area (374.9±85.4 µm 2). STG and CMG FB+ neurons were immunoreactive (IR) for tyrosine hydroxylase (TH) (66±10.1% and 52.7±8.2%, respectively), dopamine beta-hydroxylase (DbH) (62±6.2% and 52±6.2%, respectively), neuropeptide Y (NPY) (59±8.2% and 65.8±7.3%, respectively), calcitonin-gene-related peptide (CGRP) (24.1±3.3% and 22.1±3.3%, respectively), substance P (SP) (21.6±2.4% and 37.7±7.5%, respectively), vasoactive intestinal polypeptide (VIP) (18.9±2.3% and 35.4±4.4%, respectively), neuronal nitric oxide synthase (nNOS) (15.3±2% and 32.9±7.7%, respectively), vesicular acetylcholine transporter (VAChT) (15±2% and 34.7±4.5%, respectively), leuenkephalin (LENK) (14.3±7.1% and 25.9±8.9%, respectively), and somatostatin (SOM) (12.4±3% and 31.8±7.3%, respectively). UBT-projecting neurons were also surrounded by VAChT-, CGRP-, LENK-, and nNOS-IR fibers. The possible role of these neurons and fibers in the neural pathways of the UBT is discussed. We have chosen the domestic pig as experimental animal because it has been regarded as a more suitable model for studying human lower urinary tract innervation than rodents or carnivores. 15-18 Materials and Methods All the procedures described below were carried out in accordance with the European Communities Council Directive of 24 November 1986 (86/609/EEC) and the Italian legislation regarding experimental animals, after approval by the Scientific Ethics Committee for Experiments on Animals of the University of Parma (Prot. Rif. 68/09). All possible efforts were made to minimize the number of animals used and their suffering. Four intact male crossbreed (Large white x Landrace x Duroc) pigs (aged 3 months, mean body weight: 34±3 Kg, range 30-38 Kg) were maintained on a diet and water ad libitum for 1 week before the experiment. For 24 h prior to surgery, the animals were not given any food and preventive antibiotic therapy with Ceftiofur (Naxcel 5 mg/Kg i.m., Pfizer, Sandwich Kent, UK) was administered. The animals were sedated by intramuscular injection of azaperone (4-10 mg/10 Kg,
Qualitative and quantitative morphology of renal nerves in C57BL/6J mice
2002
The detailed morphology of the renal nerves in mice has not been reported previously. The aims of this study were to describe the general morphology of the extrinsic renal nerve in C57BL/6 mice, and determine its morphometric parameters. The major renal nerve innervating the left kidney was isolated in five mice. Thin sections of the nerve segments were then examined by transmission electron microscopy. The renal nerve averaged 35.4 Ϯ 3.6 (S.E.M.) m in diameter and 741 Ϯ 104 m in area. The renal nerve contained an average of 830 Ϯ 169 unmyelinated fibers and only 4.6 Ϯ 1.7 myelinated fibers. The axon diameter of myelinated and unmyelinated fibers averaged 2.2 Ϯ 0.3 m and 0.76 Ϯ 0.02 m, respectively. The diameter of the unmyelinated fibers ranged from 0.3 to 2.0 m, and the distribution histogram was unimodal. The majority of fibers (85%) had diameters of 0.6-1.0 m. These results are similar to those obtained for renal nerves of rats with respect to the predominance of unmyelinated fibers. However, the diameter of unmyelinated fibers is larger in rats and the distribution histogram of rat unmyelinated fibers is bimodal, in contrast to the unimodal distribution in mice. The morphological description of the renal nerves in mice provides baseline data for further investigations of the structural basis of altered autonomic reflexes. The results will be useful in analyses of genes that influence the development and structure of sympathetic and sensory innervation of the kidney in genetically manipulated mice. Anat Rec 268:399-404, 2002.
Neurourology and Urodynamics, 2008
Aims: The morphology and functional importance of the autonomic nervous system in the upper urinary tract is still not completely understood. Previous histological studies investigating the innervation of the urinary tract have mainly used conventional sections in which the three-dimensional structure of the intramural innervation is difficult to achieve. In contrast, the whole-mount preparation technique is a suitable method for visualizing the distribution of the mesh-like neuronal networks within the urinary tract. Methods: The distribution and regional variation of neurofilament (NF), tyrosine hydroxylase (TH), choline acetyltransferase (ChAT), and substance P-immunoreactive (SP-IR) neurons, as well as acetylcholinesterase (AChE) and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d)-positive neurons were investigated using whole-mount preparations of the guinea pig upper urinary tract. Results: Two distinct nervous plexuses were detected within the muscle layers containing NF, TH, ChAT, and SP-IR nerves. AChE-positive nerves were seen in all layers. Only moderate NADPH-d-positive innervation was found. Renal pelvis, upper and lower part of the ureter showed an overall increased innervation compared to the middle portion of the ureter. Ganglia were found at the pelviureteric border displaying NF and TH immunoreactivity. Conclusion: The whole-mount preparation technique provides an elegant method for assessing the three-dimensional architecture of ureteral innervation. The guinea pig upper urinary tract is richly supplied with adrenergic, cholinergic, nitrergic, and sensory nerves which suggest that the autonomous nervous system plays an important role in controlling ureteral motility and blood flow.
The Journal of Comparative Neurology, 1984
Anatomical and electrophysiological experiments have demonstrated a prominent projection from the sacral sympathetic chain via the pelvic nerve to postganglionic nerves on the surface of the urinary bladder and the large intestine of the cat. Retrograde labeling studies revealed that the pelvic nerve, which is generally believed to carry primarily parasympathetic axons, has a considerable population of sympathetic fibers originating mainly from the Sl-S3 paravertebral ganglia. The number of sympathetic neurons projecting to the pelvic nerve (2,100) was about 75% of that projecting to the pudendal nerve (2,9001, a somatic nerve which would be expected to carry a large sympathetic fiber constituent. Sympathetic neurons projecting t o the pudendal nerve were located primarily in the L6-SZ ganglia. Electrophysiological studies confirmed the presence of a sympathetic pathway from the paravertebral ganglia to the pelvic viscera. Electrical stimulation (thresholds 1.5-3 V) of the lumbar sympathetic chain evoked firing in the pelvic nerve and in postganglionic nerves on the surface of the colon and bladder at latencies of 60-150 msec. The responses were unaffected by cutting the chain one segment rostra1 to the site of stimulation, but were abolished by the administration of a ganglionic-blocking agent (tetraethylammonium). The responses on the colon and bladder postganglionic nerves were also abolished by transection of the pelvic nerve. The conduction velocity in the sympathetic postganglionic axons was approximately 1 &second.
BJU International, 2008
The wall of the LUT in the region immediately between the bladder base and the urethra, the BUJ, differed in its cellular composition relative to the adjacent areas. PGP-positive (PGP +) nerve fibres, presumptive afferent fibres, lay within the urothelium running between the epithelial cells. There were two general nerve patterns: branching fibres with no varicosities, and complex fibres with varicosities. Fibre collaterals with varicosities exited the urothelium and occupied the space under the urothelium adjacent to the layer of suburothelial ICs. The latter, lamina propria and around the muscle bundles were identified using vimentin (vim +). In the base a few vim + cells were also PGP +. In the region of the BUJ there was a decrease in the amount of smooth muscle. In this region, below the lamina propria, there was an area densely populated with vim + /PGP + ICs. Nerve fibres ran between the cells in this region. CONCLUSION These structural specialisations within the urothelium and deeper layers of the BUJ suggest that they might be associated with specific functions. The localized highly branched network of the putative afferent nerves suggests the presence of a local axonal reflexes involving possible cross-talk between the urothelium and suburothelial layer. The function of the specialized region of ICs is not known and must await further information on the functional properties of this novel cell type. These observations show further the cellular heterogeneity of the cells in the LUT and the complexity of the structures. One of the major current challenges in functional urology is to understand the relationships between these novel structures and overall bladder and urethral function. KEYWORDS sensory nerves, interstitial cells, bladder urethral junction OBJECTIVE To identify and characterize possible structural specialisations in the wall of the lower urinary tract (LUT) in the region of the bladder urethral junction (BUJ), with the specific objective of identifying regional variations in sensory nerve fibres and interstitial cells (ICs). MATERIALS AND METHODS The bladder base and urethra was removed from five male guinea pigs killed by cervical dislocation. Tissue pieces were incubated in Krebs' solution at 36 ° C, gassed with 95% O 2 and 5% CO 2 , fixed in 4% paraformaldehyde and processed for immunohistochemistry. The nonspecific marker vimentin and the general neuronal marker protein gene product (PGP) 9.5 were used to identify ICs and nerve fibres, respectively. Specific antibody binding was visualized using the appropriate secondary antibodies.
Early innervation of the metanephric kidney
Development, 1988
During kidney differentiation, the nephrogenic mesenchyme converts into renal tubules and the ureter bud branches to form the collecting system. Here we show that in the early undifferentiated kidney rudiment there is a third cell type present. In whole-mount preparations of cultured undifferentiated metanephric kidneys, neurones can be detected by immunohistochemical means with antibodies against the neurofilament triplet, 13AA8, and against neuronal cell surface gangliosides, Q211. Clusters of neuronal cell bodies can be seen in the mesenchyme close to the ureter bud. The terminal endings of neurites are found around the mesenchymal condensates that later become kidney tubules. A similar distribution of neurites can be revealed in tissue sections of kidney grafts growing in the chicken chorioallantoic membranes. In primary cultures of the ureter bud cells, neurones are con-stantly present. In another report, we have shown that, in experimental conditions, neurones are in-volved in...
The conduction velocities and spinal projections of single renal afferent fibers in the rat
Brain Research, 1987
This study was designed to examine the conduction velocities and spinal projections of renal afferent fibers in the rat using electrophysiological techniques. In chloralose-anesthctized rats, we electrically stimulated the peripheral ends of cut. lower thoracic and upper lumbar dorsal roots and recorded and averaged antidromically comhwted action potentials in the renal nerves. Of 284 single axons responding to stimulation of ipsilater:d dorsal roots T,~-L t, the majority were activated by stimulating roots T~t-Ti3. No antidromic responses could be elicited by stimulating the contralateral dorsal roots. Afferent fibers woe divlq~ ie into two groups, distinguished by their conduction velocities: a population of slowly conducting axons, presumably composed ot both unmyelinated (0.3-2 m/s, 76%) and thinly myelinated (2-9 m/s, 19~) fibers, and a population of more rapidly conducting, small myelinated axons (12-32 m/s, 5%). Slowly and more rapidly conducting fibers were not differentially distributed among dorsal roots. Postexperimental histological examination of nerves revealed small myelinated axons with diameters appropriate for some, but not for all, of the axons with conduction velocities in the myelimlted range. These results indicate that single myelinated and unmyelinated primary afferent axons can be identiffed by antidromic stimulation in autonomic nerves of rat. They provide the first electrophysiological description of afferent renal nerve fibers in the rat, and they verify the predominantly unmyelinated nature of these fibers.