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In this investigation we have combined the methods of ultrastructural demonstration of acetylcholinesterase activity with electron microscopic autoradiography for the demonstration of norepinephrine uptake. The results show electron-dense deposits indicative of acetylcholinesterase activity associated with perivascular axons overlaid by concentrations of silver grains representing exogenous tritiated norepinephrine. Forty-five percent of the intervaricose regions and 19% of the varicosities overlaid by autoradiographic grains showed "moderate" amounts of cholinesterase staining. A greater proportion of autoradiographic grains was observed on the varicosities than in the intervaricose regions; however, the amount of acetylcholinesterase activity was greater in the intervaricose regions than in the varicosities. This investigation provides evidence for the presence of periaxonal acetylcholinesterase staining in adrenergic axons in the rat kidney.
Anat Rec 1983 Feb
PMID:Simultaneous ultrastructural visualization of acetylcholinesterase activity and tritiated norepinephrine uptake in renal nerves. 684 69

The noradrenergic sympathetic innervation of the penis of control and 4-month streptozotocin-diabetic rats was examined with the glyoxylic acid histofluorescence method. Noradrenergic varicosities were found in the corpora cavernosa in a dense subtunical plexus and in the perisinusoidal and trabecular regions of the erectile tissue, in the corpus spongiosum in perisinusoidal tissue, around large arteries and veins, and around small tortuous arterioles and small draining veins of the corpora cavernosa and spongiosum. Noradrenergic varicosities were diminished in number and fluorescent intensity in all regions of the penis of diabetic rats compared with controls. The subtunical plexus was absent, perisinusoidal and trabecular varicosities were sparse, and only occasional intermittent, discontinuous, dull fluorescent fibers or plexuses were found around the vessels. Quantitation with high-performance liquid chromatography revealed a significant reduction of norepinephrine in the penis of diabetic rats compared with controls. The present study suggests that long-term streptozotocin diabetes in the rat is accompanied by sympathetic autonomic neuropathy of the penis that seems to parallel changes in the noradrenergic content of penile corpora of men with diabetes and erectile impotence. The streptozotocin-diabetic rat merits further study to explore the relationship between noradrenergic innervation of the penis and erectile tissue.
Anat Rec 1983 May
PMID:Noradrenergic innervation of the penis in control and streptozotocin-diabetic rats: evidence of autonomic neuropathy. 688 50

Lamprey, Entosphenus japonicus, cerebral blood vessel autonomic nerve supply was studied with fluorescence and cholinesterase histochemistry and electron microscopy. Nerve fibers emitting a yellow fluorescence characteristic of serotonin (Exc./Em. max.; 380/530 nm) were found on the major cerebral and pial arteries, but not acetylcholinesterase (AChE)-positive ones. Single ganglion cells also emitting a strong yellow fluorescence were seen in the artery adventitia. On rare occasions these cells were observed in pairs. Terminal varicosities of central catecholamine-containing nerves (Exc./Em. max.; 410/475 nm) were observed on parenchymal capillaries, but not central AChE-positive nerve terminals. In ganglion cells, dense cored vesicles (ca. 130 nm in average diameter; DCV) were abundant in the Golgi area, suggesting their formation at this site. Two types of DCV were observed; one with a homogeneous dense core and the other with a granular core. DCV were numerous in axons as well, axons in which many small clear vesicles (40--60 nm in diameter) as well as DCV were occasionally observed. The question of whether the small clear vesicles or the DCV contained serotonin could not be resolved.
Anat Rec 1980 Dec
PMID:A histochemical and ultrastructural study of serotonin-containing nerves in cerebral blood vessels of the lamprey. 721 16

The histological structure of the gerbil Harderian gland was investigated by means of light and transmission electron microscopy. The single excretory duct of the gland is directly continuous with endpieces at the hilus and opens nasally and ventrally to the third eyelid. The excretory duct is accompanied by many acini of small serous glands around it. The gland is composed of tubuloalveoli (tubular alveoli) with wide lumina and is not divided into lobules. There is no branched duct system within the gland. The tubuloalveoli themselves convey the secretory materials to the hilus where the excretory duct begins. The alveolar epithelium is composed of only one type of glandular cell as well as myoepithelial cells. The glandular cells contain many clear secretory vacuoles containing lipids and well-developed tubular smooth endoplasmic reticulum. The secretory vacuoles are surrounded by a unit membrane and are secreted by exocytosis. The interstices of the gland contain two types of autonomic nerve varicosities and a number of melanocytes. The surface of the gland is covered with the endothelium of the orbital venous sinus.
Anat Rec 1981 Jul
PMID:A histological study of the Harderian gland of Mongolian gerbils, Meriones meridianus. 727 Sep 26

The pelvic flexure portion of the equine large colon is the proposed location of a pacemaker mechanism. This study was conducted to ascertain whether the distribution of certain putative neurotransmitters differs at the pelvic flexure compared to other sampling sites. Tissue samples were collected from the intestinal tracts of six horses. Serial sections from these samples were reacted with primary antisera specific for substance P, vasoactive intestinal polypeptide (VIP), methionine-Enkephalin, and calcitonin gene-related peptide (CGRP). The regional distribution of immunoreactive neuronal elements was uniform for each of the neuropeptides except VIP. Although neurons exhibiting VIP-like immunoreactivity were abundant throughout the colon, they were somewhat more plentiful near the apex of the pelvic flexure and the left dorsal colon. These neurons may participate in the initiation and propagation of the propulsive/retropulsive contraction waves, which emanate from this location and are believed to lend a sphincter-like capacity to the pelvic flexure. The submucosal plexus was replete with neurons with intense substance P and VIP-like reactivity. Reactive fibers left submucosal ganglia to project to the intestinal mucosa, reflecting a possible secretogogic role for these neurons. This role may be especially important for the horse as a hindgut fermenter. There were abundant methionine-Enkephalin and substance P-like reactive varicosities throughout the myenteric plexus, many of which established a pericellular plexus of varicose fibers. The abundance of these varicosities, which may correlate with a high degree of neuronal integration, did not vary regionally. These data may enhance our understanding of both normal colonic peristalsis and motility disorders caused by a depletion of these neuropeptides.
Anat Rec 1993 Jun
PMID:Neuropeptide distributions in the colon, cecum, and jejunum of the horse. 768 32

Despite extensive knowledge of the neuroepithelial endocrine (NEE) system in the lungs of species of various vertebrate classes, data on avians are limited. The present investigation deals with the light- and electron-microscopical immunocytochemistry and morphology of pulmonary NEE cells in the quail, Coturnix coturnix. Light-microscopically, serotonin immunoreactivity was detected in numerous solitary and clustered NEE cells located in the cilio-mucous epithelium of primary and secondary bronchi in adult as well as in newly hatched quails. Only in newly hatched quails could a small number of bombesin- and somatostatin-like immunoreactive NEE cells be demonstrated. Electron-microscopical morphology revealed that NEE cells contained dense-cored vesicles of a wide range of diameters and electron densities. Nearly all of the NEE cells were seen to rest on the basement membrane of the cilio-mucous epithelium, lacking direct contact with the luminal surface. Nerve varicosities or nerve endings, of both afferent and efferent morphological appearance, were found directly apposed to the basal portion of NEE cells, invaginating between NEE cells or between NEE cells and adjacent epithelial cells. Often, synaptic specializations could be recognized between NEE cells and nerve terminals. Electron-microscopical immunocytochemistry confirmed that the intraepithelial serotonin-containing cells correspond to the cells with NEE characteristics. Moreover, two types of NEE cells could be distinguished in newly hatched quail lungs. Both types showed serotonin immunoreactivity selectively distributed over the dense-cored vesicles, but somatostatin- and bombesin-like immunoreactivities were only noted in one of the NEE cell types and were never seen colocalized. Thus, the avian NEE system too, harbors at least three different bioactive substances and has a morphology comparable to that of mammals, reptiles and amphibians.
Anat Rec 1994 May
PMID:The pulmonary neuroepithelial endocrine system in the quail, Coturnix coturnix. Light- and electron-microscopical immunocytochemistry and morphology. 791 90

Much physiologic and morphologic research has been done into the sensory mechanism of the frog muscle spindle. However, no freeze-fracture study has described in detail the shape and intramembrane structure of the nonmyelinated sensory axon terminals of the frog muscle spindle. In this study, muscle spindles were isolated from the red part of bullfrog semitendinous muscles. Chemically fixed spindles were subjected to freeze fracturing. The sensory axon endings were reconstructed, and the size and density of intramembrane particles (IMPs) were measured along the sensory nerve endings. The axon terminals had four distinctive parts: parent trunks (>0.5 microm in diameter), primary branches (0.15-0.5 microm), terminal branches (<0.1 pm), and varicosities (0.02-0.5 microm). IMPs ranged from 5 nm to 21 nm in diameter and were present in the intramembrane space of the plasma membrane all throughout the nonmyelinated sensory nerve endings. Mean IMP sizes in the protoplasmic face (PF) and the external face (EF), respectively, were 8.1 nm and 8.4 nm in the parent trunks, 8.8 nm and 8.8 nm in the primary branches, 9.4 nm and 9.0 nm in the varicosities, and 8.7 nm and 8.7 nm in the terminal branches. Mean IMP size in the PF was smallest in the parent trunk and largest in the varicosity. Mean IMP densities (numbers of IMPs per microm2) in the PF and the EF, respectively, were 2,500 and 700 in the parent trunks, 2,200 and 500 in the primary branches, 1,700 and 400 in the varicosities, and 1,000 and 300 in the terminal branches. Density decreased with the tapering of the axon terminal, with IMPs distributed evenly in the PF and the EF. The characteristic intramembrane structure of sensory nerve endings is discussed.
Anat Rec 1998 11
PMID:Intramembrane structure of the sensory axon terminals in bullfrog muscle spindles. 981 Dec 12

The cholinergic and histaminergic projections have important neuromodulatory functions in the ascending visual pathways, so we compared the pattern and mode of innervation of the two projections in the lateral geniculate complex (dorsal lateral geniculate nucleus and pregeniculate nucleus) of the macaque monkey. Brain tissue from macaques was immunoreacted by means of antibodies to choline acetyltransferase (ChAT) or to histamine and processed for light and electron microscopy. A dense plexus of thin, highly branched ChAT-immunoreactive axons laden with varicosities was found in all layers of the dLGN including the koniocellular laminae and in the pregeniculate nucleus. ChAT label was more dense in magnocellular layers 1 and 2 than in parvocellular layers 3-6 and relatively sparse in the interlaminar zones. Varicosities associated with the cholinergic axons had an average of three conventional asymmetric synapses per varicosity, and these appeared to contact dendrites of both thalamocortical cells and interneurons. Histamine-immunoreactive axons were distributed homogeneously throughout all laminar and interlaminar zones of the dLGN, but were denser in the pregeniculate nucleus than in the dLGN. Histaminergic axons branched infrequently and were typically larger in caliber than cholinergic axons. The overwhelming majority of varicosities were found en passant and rarely displayed conventional synapses, despite the abundance of synaptic vesicles, and were not associated preferentially with specific cellular structures. The innervation of the macaque dLGN complex by cholinergic and histaminergic systems is consistent with their proposed role in state dependent modulation of thalamic activity. The dense and highly synaptic innervation by cholinergic axons supports the proposal of additional involvement of these axons in functions related to eye movements.
Anat Rec 1999 07 01
PMID:Comparison of cholinergic and histaminergic axons in the lateral geniculate complex of the macaque monkey. 1041 96

Histochemical staining techniques for 5'-nucleotidase (5'-Nase) and acetylcholinesterase (AChE) were undertaken to localize the lymphatic network and nerve plexus in the monkey urinary bladder. Abundant 5'-Nase-positive lymphatic networks were characterized by increased number of valve-like structures and decreased calibre of blind-ends from the subepithelium to the subserosa. AChE-positive nerve fibers were visible throughout the vesical walls as fine plexuses, the densest being the neuromuscular plexus among the detrusor muscle cells or in each muscle bundle. AChE-positive nerve fibers or terminals were more frequently discernible around blood vessels than around lymphatics, and showed more intimate association with the lymphatics in the muscularis than those in the subepithelium. The nerve terminals in the subepithelium were frequently separated from attenuated lymphatic endothelium by the long processes of fibroblasts or some connective tissue cells. An ultrastructural observation revealed that unmyelinated nerve fibers with numerous neurofilaments and neurotubules run in close apposition to the lymphatic endothelium. Noteworthily, fewer terminal varicosities containing numerous small agranular vesicles (30-50 nm) and mitochondria, partially or completely bare of their Schwann cell covering in the vicinity of the lymphatic endothelium, were found in the subendothelium of initial lymphatics than in collecting ones. These terminals were occasionally identified at a distance of 120-350 nm from the subendothelial aspect of valve-originating roots, although no direct innervation of the vascular muscle cells could be found. A loose fibro-elastic connective tissue was usually interlaced between glial cell covering and lymphatic endothelium. The intrinsic interrelation of the lymphatic wall with the nerve plexus implies that the twisted subendothelial nerve terminals might be involved in intramural lymph drainage of the bladder.
Anat Rec 2000 05 01
PMID:Intrinsic interrelation of lymphatic endothelia with nerve elements in the monkey urinary bladder. 1076 Jul 47

Lower numbers of neuropeptide-containing fibers in arthritic joints have been found as compared to control joints. This may be the result of fiber depletion, necrosis of fibers, or proliferation of soft tissues without neural sprouting. To discriminate between these possibilities, we studied the relationships between soft tissue proliferation, changes in vascularity of synovial tissues, and changes in joint innervation during arthritis. Arthritis was induced in the knee joint of mice by a single subpatellar injection of methylated bovine serum albumin after previous immunization. Antibodies to protein gene product 9.5, S-100, and growth-associated protein-43 (GAP-43) were used to study the general innervation pattern. Antibodies to calcitonin gene-related peptide (CGRP), vasointestinal polypeptide (VIP), substance P (SP), and tyrosine hydroxylase (TH) were used to localize sensory (SP, CGRP, VIP) and sympathetic (TH) fibers. Blood vessels of the joint were studied with ink perfusion, GAP-43, and a vascular marker (LF1). Directly after the induction of arthritis, the synovial cavity was enlarged and filled with leukocytes. From day 4 onward, small sprouting blood vessels penetrated the avascular mass of cells in the joint cavity. After 1 week, the vascular sprouting activity and GAP-43 immunoreactivity were maximal, and after 2 weeks, vascular sprouting activity diminished. In the subsequent period, the synovia slowly regained their prearthritic appearance and thickness. The most pronounced changes in the general staining pattern of CGRP, SP, VIP, and TH were found in the periosteum. From 2 days to 4 weeks after the induction of arthritis, the layer of SP, CGRP, and VIP fibers in the femoral periosteum was thicker and more irregular. GAP-43 staining showed many terminal varicosities, which suggested sprouting of nerve fibers. From 2 days to 2 weeks after the induction of arthritis, the SP and CGRP fibers in the periosteum showed gradual depletion. In the thickened subsynovial tissues that were revascularized, no ingrowth of neural elements was found. As the total number of nerve fibers in the synovial tissue did not change, large parts of the synovia directly facing the joint cavity were not innervated at 1 week after the induction of arthritis. These results strongly suggest that periosteal SP and CGRP fibers were depleted during arthritis. Synovial proliferation without concomitant fiber growth is the main cause of the reduced number of immunocytochemically detectable fibers in the mouse arthritic knee joint.
Anat Rec 2000 09 01
PMID:Neurovascular plasticity in the knee joint of an arthritic mouse model. 1096 36


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