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1.
Auton Neurosci ; 156(1-2): 152-3, 2010 Aug 25.
Article in English | MEDLINE | ID: mdl-20537599

ABSTRACT

A dense network of nerves containing neuronal nitric oxide synthase is present in cerebral vessels from experimental animals. The nerves may regulate cerebrovascular tone, protect the brain from stroke, and contribute to cluster headaches in humans; but studies in humans have shown only modest nitroxidergic innervation of cerebral vessels. We tested the hypothesis that nerve fibers containing neuronal nitric oxide synthase richly innervate human cerebral arteries. We used immunohistochemical techniques at post mortem and found dense neuronal nitric oxide synthase nerve staining in human cerebral vessel walls consistent with participation of nitroxidergic fibers in human physiological and pathophysiological processes.


Subject(s)
Cerebral Arteries/enzymology , Cerebral Arteries/innervation , Nitric Oxide Synthase Type I/metabolism , Nitric Oxide/physiology , Parasympathetic Fibers, Postganglionic/enzymology , Humans , Male , Middle Aged
2.
J Clin Neurosci ; 15(9): 1023-6, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18617399

ABSTRACT

Sympathetic catecholaminergic fibers in the vagus nerve were immunohistochemically examined in formalin-fixed human cadavers using an antibody against the noradrenalin-synthetic enzyme tyrosine hydroxylase (TH). TH-positive fibers were extensively distributed in the vagal nerve components, including the superior and inferior ganglia, the main trunk and the branches (superior and recurrent laryngeal, superior and inferior cardiac, and pulmonary branches). The inferior ganglion and its continuous cervical main trunk contained numerous TH-positive fibers with focal or diffuse distribution patterns in each nerve bundle. From these findings, we conclude that sympathetic fibers are consistently included in the human vagus nerve, a main source of parasympathetic preganglionic fibers to the cervical, thoracic and abdominal visceral organs.


Subject(s)
Adrenergic Fibers/ultrastructure , Catecholamines/biosynthesis , Tyrosine 3-Monooxygenase/metabolism , Vagus Nerve/cytology , Vagus Nerve/enzymology , Adrenergic Fibers/enzymology , Aged , Aged, 80 and over , Female , Ganglia, Sensory/cytology , Ganglia, Sensory/enzymology , Ganglia, Sympathetic/cytology , Ganglia, Sympathetic/enzymology , Humans , Immunohistochemistry , Laryngeal Nerves/cytology , Laryngeal Nerves/enzymology , Male , Middle Aged , Parasympathetic Fibers, Postganglionic/cytology , Parasympathetic Fibers, Postganglionic/enzymology , Tyrosine 3-Monooxygenase/analysis , Viscera/innervation
3.
J Appl Physiol (1985) ; 98(5): 1792-8, 2005 May.
Article in English | MEDLINE | ID: mdl-15626754

ABSTRACT

Effects of electrical stimulation of the nerve bundles including sensory and parasympathetic nerves innervating cerebral arteries on cerebral blood flow (CBF) and mean arterial blood pressure (MABP) were investigated with a laser-Doppler flowmeter and a blood pressure monitoring system in anesthetized rats pretreated with and without capsaicin. The electrode was hooked on the nerve bundles including the distal nasociliary nerve from trigeminal nerve and parasympathetic nerve fibers from sphenopalatine ganglion. In control rats, the nerve stimulation for 30 s increased CBF in the ipsilateral side and MABP. Hexamethonium attenuated the increase in CBF and abolished that in MABP. Under treatment with hexamethonium, N(G)-nitro-L-arginine (L-NNA, 1 mg/kg) significantly attenuated the stimulation-induced increase in CBF, which was restored by the addition of L-arginine. Although the dose of L-NNA was raised up to 10 mg/kg, the stimulation-induced increase in CBF was not further inhibited and was never abolished. In capsaicin-pretreated rats, magnitudes of the stimulation-induced increases in CBF and MABP were lower than those in control rats. Hexamethonium attenuated the increase in CBF and abolished that in MABP. Under treatment with hexamethonium, L-NNA abolished the stimulation-induced increase in CBF in capsaicin-pretreated rats. In conclusion, nitric oxide released from parasympathetic nerves and neuropeptide(s) released antidromically from sensory nerves may be responsible for the increase in CBF in the rat. The afferent impulses by nerve stimulation may stimulate the trigeminal nerve and lead to the rapid increase in MABP, which partly contributes to the increase in CBF.


Subject(s)
Capsaicin/pharmacology , Cerebrovascular Circulation/drug effects , Neurons, Afferent/drug effects , Nitric Oxide Synthase/antagonists & inhibitors , Parasympathetic Fibers, Postganglionic/drug effects , Animals , Cerebrovascular Circulation/physiology , Electric Stimulation/methods , Enzyme Inhibitors/pharmacology , Male , Neurons, Afferent/enzymology , Nitric Oxide Synthase/metabolism , Parasympathetic Fibers, Postganglionic/enzymology , Rats , Rats, Wistar
4.
Neuroscience ; 125(3): 663-70, 2004.
Article in English | MEDLINE | ID: mdl-15099680

ABSTRACT

The aim of this study was to examine the participation of nitrergic neurotransmission in the initiation of micturition hyperreflexia associated to cyclophosphamide (CP)-induced cystitis in rats. Micturition threshold volume was significantly reduced 4 h after CP administration (100 mg/kg, i.p.); this reduction was attenuated by intra-arterially injected N(G)-nitro-l-arginine-methyl ester (l-NAME), a non selective nitric oxide synthase (NOS) inhibitor, but not by intravesical infusion of S-methyl-l-thiocitrulline (l-SMTC), another structurally different NOS inhibitor. Interestingly, l-NAME failed to affect micturition threshold volume in normal rats. The magnitude of isolated detrusor strips contractions elicited by either carbachol or nerve activation was significantly reduced in CP-treated rats but was unaffected by the addition of N(G)-nitro-l-arginine (l-NOARG), a nonselective NOS inhibitor. In contrast, intrathecal l-NAME and l-SMTC but not N(G)-nitro-d-arginine-methyl ester (d-NAME) administration augmented the micturition threshold volume in CP-treated rats in an l-arginine preventable manner. As with the systemic injection, intrathecal l-NAME also did not affect the micturition threshold volume in normal rats. Four hours after CP injection, the number of neuronal NOS immunoreactive or nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) positive neurons in spinal lumbosacral segments (L6-S2) was not altered whereas the number of c-Fos immunoreactive neurons increased significantly in the dorsal gray commissural nucleus (DGC), the parasympathetic sacral nucleus (PSN) and lamina X of these segments. Ca(2+)-dependent, but not Ca(2+)-independent NOS activity increased significantly in spinal L6-S2 segments but not in thoracic segments of CP-treated rats. These data indicate that the micturition hyperreflexia observed in the initial hours of CP-induced cystitis is associated with an increase in Ca(2+)-dependent NOS activity in spinal L6-S2 segments suggesting an increased production of nitric oxide (NO). The increased production of NO in these spinal segments appears to be necessary for the initiation of the micturition hyperreflexia.


Subject(s)
Cystitis/complications , Nitric Oxide Synthase/metabolism , Parasympathetic Fibers, Postganglionic/enzymology , Spinal Cord/enzymology , Urinary Bladder/innervation , Urinary Bladder/physiopathology , Urination Disorders/enzymology , Animals , Cyclophosphamide , Cystitis/chemically induced , Cystitis/physiopathology , Enzyme Inhibitors/pharmacology , Female , Immunohistochemistry , Injections, Spinal , Muscle, Smooth/drug effects , Muscle, Smooth/enzymology , NADPH Dehydrogenase/metabolism , NG-Nitroarginine Methyl Ester/pharmacology , Neurons/cytology , Neurons/drug effects , Neurons/enzymology , Nitric Oxide/biosynthesis , Nitric Oxide Synthase/antagonists & inhibitors , Parasympathetic Fibers, Postganglionic/physiopathology , Proto-Oncogene Proteins c-fos/drug effects , Proto-Oncogene Proteins c-fos/metabolism , Rats , Rats, Wistar , Reflex, Abnormal/drug effects , Spinal Cord/cytology , Spinal Cord/physiopathology , Urination/drug effects , Urination/physiology , Urination Disorders/etiology , Urination Disorders/physiopathology
5.
J Chem Neuroanat ; 22(3): 147-55, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11522437

ABSTRACT

It is generally considered that parasympathetic postganglionic nerve fibers innervating the lacrimal gland (LG) arise from the pterygopalatine ganglion (PPG), while sympathetic and sensory innervations arise from the superior cervical ganglion (SCG) and trigeminal ganglion (TG), respectively. Recently, we reported for the first time that the parasympathetic innervation of the cat LG was also provided by the otic ganglion (OG) and ciliary ganglion (CG), and that the sensory innervation was also provided by the superior vagal ganglion (SVG) and superior glossopharyngeal ganglion (SGG). To determine if nitric oxide (NO) is a neurotransmitter of the autonomic and sensory neurons innervating the LG, we injected the cholera toxin B subunit (CTB) as a retrograde tracer into the cat LG, and used double-labeling fluorescent immunohistochemistry for CTB and nitric oxide synthase (NOS). We found that NOS-/CTB-immunofluorescent double-labeled perikarya were localized in the PPG, OG, TG, SVG and SGG, but not in the CG and SCG. The highest numbers of NOS-/CTB-immunofluorescent double-labeled neurons were found in the PPG and TG. In addition, we examined the presence of nitrergic nerve fibers in the LG using NADPH-d histochemistry and found that a large amount of NADPH-d-stained nerve fibers were distributed around the glandular acini and in the walls of glandular ducts and blood vessels. This study provides the first direct evidence showing that NO may act as a neurotransmitter or modulator involved in the parasympathetic and sensory regulation of lacrimal secretion and blood circulation, but may not be implicated in the sympathetic control of LG activities, and that nitrergic nerve fibers in the LG arise mainly from parasympathetic postganglionic neurons in the PPG and sensory neurons in the TG. The present results suggest that NO plays an important role in the regulation of LG activities.


Subject(s)
Ganglia, Parasympathetic/enzymology , Ganglia, Sensory/enzymology , Lacrimal Apparatus/enzymology , Lacrimal Apparatus/innervation , Neurons/enzymology , Nitric Oxide Synthase/metabolism , Parasympathetic Fibers, Postganglionic/enzymology , Animals , Cats , Cell Count , Cholera Toxin/pharmacokinetics , Female , Fluorescent Antibody Technique , Ganglia, Parasympathetic/cytology , Ganglia, Sensory/cytology , Ganglia, Sympathetic/cytology , Ganglia, Sympathetic/enzymology , Glossopharyngeal Nerve/cytology , Glossopharyngeal Nerve/enzymology , Lacrimal Apparatus/cytology , Male , NADPH Dehydrogenase/metabolism , Neurons/cytology , Nitric Oxide/metabolism , Parasympathetic Fibers, Postganglionic/cytology , Sympathetic Fibers, Postganglionic/cytology , Sympathetic Fibers, Postganglionic/enzymology , Trigeminal Ganglion/cytology , Trigeminal Ganglion/enzymology , Vagus Nerve/cytology , Vagus Nerve/enzymology
6.
J Neurophysiol ; 79(6): 3216-28, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9636120

ABSTRACT

Trimming all but two whiskers in adult rats produces a predictable change in cortical cell-evoked responses characterized by increased responsiveness to the two intact whiskers and decreased responsiveness to the trimmed whiskers. This type of synaptic plasticity in rat somatic sensory cortex, called "whisker pairing plasticity," first appears in cells above and below the layer IV barrels. These are also the cortical layers that receive the densest cholinergic inputs from the nucleus basalis. The present study assesses whether the cholinergic inputs to cortex have a role in regulating whisker pairing plasticity. To do this, cholinergic basal forebrain fibers were eliminated using an immunotoxin specific for these fibers. A monoclonal antibody to the low-affinity nerve growth factor receptor 192 IgG, conjugated to the cytotoxin saporin, was injected into cortex to eliminate cholinergic fibers in the barrel field. The immunotoxin reduces acetylcholine esterase (AChE)-positive fibers in S1 cortex by >90% by 3 wk after injection. Sham-depleted animals in which either saporin alone or saporin unconjugated to 192 IgG is injected into the cortex produces no decrease in AChE-positive fibers in cortex. Sham-depleted animals show the expected plasticity in barrel column neurons. In contrast, no plasticity develops in the ACh-depleted, 7-day whisker-paired animals. These results support the conclusion that the basal forebrain cholinergic projection to cortex is an important facilitator of synaptic plasticity in mature cortex.


Subject(s)
Autonomic Pathways/physiology , Neuronal Plasticity/physiology , Parasympathetic Nervous System/physiology , Prosencephalon/physiology , Somatosensory Cortex/physiology , Acetylcholine/physiology , Acetylcholinesterase/metabolism , Animals , Autonomic Pathways/enzymology , Electrophysiology , Histocytochemistry , Male , Microelectrodes , Motor Cortex/enzymology , Motor Cortex/physiology , Parasympathetic Fibers, Postganglionic/enzymology , Parasympathetic Fibers, Postganglionic/physiology , Parasympathetic Nervous System/enzymology , Physical Stimulation , Prosencephalon/enzymology , Rats , Somatosensory Cortex/enzymology , Vibrissae/physiology
7.
Neuroscience ; 83(2): 607-16, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9460767

ABSTRACT

Cross-sections of the vas deferens taken from control adult male rats showed positive histochemical reactivity to acetylcholinesterase and immunoreactivity for antibodies to protein gene product 9.5, tyrosine hydroxylase, neuropeptide Y, vasoactive intestinal polypeptide, nitric oxide synthase and calcitonin gene-related peptide. Immunoreactivity to substance P was very sparse. Histochemical reactivity to acetylcholinesterase and immunoreactivity to vasoactive intestinal polypeptide and nitric oxide synthase was concentrated in the subepithelial lamina propria and inner smooth muscle layers. Complete surgical denervation resulting from transection of the nerve arising from the pelvic ganglion which supplies the vas deferens totally abolished the immunoreactivity to all of the antibodies tested as well as the histochemical reactivity to acetylcholinesterase. In sections of the prostatic end of the vas deferens taken from rats neonatally pretreated with capsaicin, immunoreactivity to calcitonin gene-related peptide and substance P was reduced by 75 and 83%, respectively. Immunoreactivity to neuropeptide Y, vasoactive intestinal polypeptide and nitric oxide synthase was similar in tissue sections taken from capsaicin-treated rats and those taken from control tissues. Pretreatment of rats with guanethidine or 6-hydroxydopamine decreased immunoreactivity to tyrosine hydroxylase and neuropeptide Y by 60-70%, but immunoreactivity to substance P, vasoactive intestinal polypeptide and nitric oxide synthase was unchanged, while immunoreactivity to calcitonin gene-related peptide and acetylcholinesterase staining was increased by guanethidine but not by 6-hydroxydopamine treatment. Triple labelling experiments showed nitric oxide synthase, vasoactive intestinal polypeptide and acetylcholinesterase all to be co-localized in some nerve fibres. These results indicate that the nitric oxide synthase contained in the nerve fibres innervating the rat vas deferens is unaffected by pretreatment of rats with capsaicin, 6-hydroxydopamine or guanethidine but is abolished by surgical denervation, of postganglionic parasympathetic, sympathetic and sensory nerves. Therefore it appears that nitric oxide synthase is co-localized with vasoactive intestinal polypeptide in the postganglionic parasympathetic nerves which innervate the rat vas deferens.


Subject(s)
Nitric Oxide Synthase/metabolism , Parasympathetic Fibers, Postganglionic/metabolism , Vas Deferens/innervation , Vasoactive Intestinal Peptide/metabolism , Animals , Capsaicin/pharmacology , Denervation , Guanethidine/pharmacology , Immunohistochemistry , Male , Nerve Tissue Proteins/metabolism , Oxidopamine , Parasympathetic Fibers, Postganglionic/enzymology , Rats , Rats, Sprague-Dawley , Sympatholytics/pharmacology , Thiolester Hydrolases/metabolism , Ubiquitin Thiolesterase , Vas Deferens/enzymology , Vas Deferens/metabolism
8.
J Auton Nerv Syst ; 61(3): 227-34, 1996 Dec 14.
Article in English | MEDLINE | ID: mdl-8988479

ABSTRACT

Innervation of the molar gland and blood vessels in the lower lip, gingiva and cheek mucous membrane was investigated in the cat with the aid of whole mount acetylthiocholinesterase (WATChE) histochemistry and retrograde neuronal tracing methods with horseradish peroxidase (HRP) and HRP-conjugated wheat germ agglutinin (WGA-HRP). The molar gland was found to be supplied from the buccal nerve and branches of the mylohyoid nerve on the basis of microdissection of WATChE-stained mandibular preparations under a dissecting microscope. The rostral half of the lower lip-gingiva was innervated by mental branches from the inferior alveolar nerve. The caudal half of the lower lip-gingiva and cheek mucous membrane were observed to be supplied from the buccal nerve. Following injections of HRP/WGA-HRP into the molar gland, lower lip-gingiva and cheek, many retrogradely labeled ganglion neurons were observed in the ipsilateral main and accessory otic ganglia, superior cervical ganglion and mandibular division of the trigeminal ganglion. In the pterygopalatine ganglion, a small number of positive neurons were found, but in a few cases in which the injected tracer was restricted to the lower lip-gingiva and anterior half of the molar gland, labeled neurons were not detected in the main ganglion nor in its accessory microganglia. These findings indicate that the cat molar gland receives a postganglionic parasympathetic supply from the otic ganglia, postganglionic sympathetic input from the superior cervical ganglion and sensory innervation from the trigeminal ganglion by way of the buccal nerve and mylohyoid nerve. Vessels in the rostral half of the lower lip-gingiva receive the same inputs from the inferior alveolar nerve, and vessels in the caudal half receive inputs from the buccal nerve. The vessels in the cheek mucous membrane receive dual parasympathetic supplies from the otic ganglia and the pterygopalatine ganglion by way of the buccal nerve.


Subject(s)
Autonomic Nervous System/physiology , Blood Vessels/innervation , Exocrine Glands/innervation , Neurons, Afferent/physiology , Acetylcholinesterase/metabolism , Animals , Blood Vessels/physiology , Cats , Exocrine Glands/physiology , Ganglia, Parasympathetic/cytology , Ganglia, Sympathetic/cytology , Gingiva/blood supply , Gingiva/innervation , Gingiva/physiology , Histocytochemistry , Lip/blood supply , Lip/innervation , Lip/physiology , Mouth Mucosa/blood supply , Mouth Mucosa/innervation , Mouth Mucosa/physiology , Parasympathetic Fibers, Postganglionic/enzymology , Parasympathetic Fibers, Postganglionic/physiology , Regional Blood Flow/physiology , Sympathetic Fibers, Postganglionic/enzymology , Sympathetic Fibers, Postganglionic/physiology , Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate
9.
J Comp Neurol ; 375(4): 527-51, 1996 Nov 25.
Article in English | MEDLINE | ID: mdl-8930785

ABSTRACT

The distribution of choline acetyltransferase (ChAT)-immunoreactive and acetylcholinesterase (AChE)-positive fibers and terminals was analyzed in the hippocampal formation of macaque monkeys subjected to transection of the fimbria/fornix. Cases with either unilateral or bilateral transections were prepared, with post transection survival times ranging from 2 weeks to 1.5 years. The fimbria/fornix transection resulted in a dramatic decrease in the number of cholinergic fibers in most regions of the hippocampal formation. Some hippocampal regions, however, showed relatively greater sparing of ChAT- or AChE-positive fibers. In practically all regions of the hippocampal formation, residual AChE-positive fibers were more abundant than ChAT-immunoreactive fibers. In animals with unilateral lesions, the distribution patterns and density of AChE and ChAT staining on the side contralateral to the lesion were generally similar to those of sections from unlesioned control brains. The largest decreases in the densities of positive fibers were observed in the dentate gyrus, CA3 and CA2 fields of the hippocampus, subiculum, parasubiculum, and medial and caudal parts of the entorhinal cortex. Fibers were relatively better preserved in the rostral or uncal portion of the hippocampus and dentate gyrus and in the rostral portion of the entorhinal cortex. The presubiculum demonstrated remarkable sparing that contrasted with the almost complete loss of fibers in the parasubiculum. Interestingly, animals killed approximately 1.5 years after the fornix transection showed essentially the same pattern of fiber loss as the cases with shorter survival periods. This indicates that the residual ChAT-immunoreactive fibers, many of which reach the hippocampal formation through a ventral cholinergic pathway, are not capable of reinnervating the denervated portions of the hippocampal formation. This appears to distinguish the monkey from the rat, for which substantial sprouting and reinnervation of cholinergic fibers have been reported after similar lesions.


Subject(s)
Hippocampus/physiology , Parasympathetic Nervous System/physiology , Acetylcholinesterase/metabolism , Animals , Choline O-Acetyltransferase/metabolism , Denervation , Dentate Gyrus/anatomy & histology , Dentate Gyrus/enzymology , Dentate Gyrus/physiology , Entorhinal Cortex/anatomy & histology , Entorhinal Cortex/enzymology , Entorhinal Cortex/physiology , Hippocampus/enzymology , Immunohistochemistry , Macaca fascicularis , Nerve Regeneration/physiology , Parasympathetic Fibers, Postganglionic/enzymology , Parasympathetic Fibers, Postganglionic/physiology , Parasympathetic Nervous System/enzymology , Terminology as Topic
10.
J Auton Nerv Syst ; 56(3): 207-14, 1996 Jan 05.
Article in English | MEDLINE | ID: mdl-8847445

ABSTRACT

By using mechanical nerve ligation or nerve pinch technique, we provide evidence that nitric oxide synthase (NOS) is transported in the preganglionic sympathetic axons, while postganglionic axons lack NOS transport. This finding corroborates the preganglionic sympathetic terminal as the site of NO synthesis, which is known to affect ganglionic transmission. Both vasoactive intestinal polypeptide (VIP) and substance P (SP) containing neurons of the nodose ganglion transport NOS in their axons. These results therefore suggest that NOergic innervation of autonomically innervated tissues is of parasympathetic and/or sensory, rather than sympathetic, origin.


Subject(s)
Autonomic Nervous System/enzymology , Axonal Transport/physiology , Axons/enzymology , Nitric Oxide Synthase/metabolism , Animals , Autonomic Fibers, Preganglionic/enzymology , Autonomic Fibers, Preganglionic/physiology , Autonomic Nervous System/cytology , Fluorescent Antibody Technique, Indirect , Ganglia, Parasympathetic/cytology , Ganglia, Parasympathetic/enzymology , Ganglia, Parasympathetic/physiology , Ganglia, Sympathetic/cytology , Ganglia, Sympathetic/enzymology , Ganglia, Sympathetic/physiology , Ganglionectomy , NADPH Dehydrogenase/analysis , Parasympathetic Fibers, Postganglionic/enzymology , Parasympathetic Fibers, Postganglionic/physiology , Rats , Rats, Wistar , Substance P/metabolism , Sympathetic Fibers, Postganglionic/enzymology , Sympathetic Fibers, Postganglionic/physiology , Vagus Nerve/enzymology , Vagus Nerve/physiology , Vasoactive Intestinal Peptide/metabolism
11.
Neurosci Lett ; 202(1-2): 69-72, 1995 Dec 29.
Article in English | MEDLINE | ID: mdl-8787833

ABSTRACT

Neuronal somata in the rat kidney are very often part of ganglionated plexus and contain nitric oxide synthase (NOS). Examining serial 100 microns slices of whole kidneys, we identified three subpopulations of neuronal somata by: (a) staining for NADPH-diaphorase (NADPH-d) histochemistry followed by the demonstration of dopamine beta-hydroxylase (DBH) by immunoperoxidase, and (b) staining for DBH by immunofluorescence followed by the demonstration of NADPH-d histochemical activity. The largest subpopulation of neuronal somata displayed both DBH immunoreactivity and NADPH-d histochemical activity. The second largest group of somata showed NADPH-d activity only. A small group of neuronal somata showed only DBH immunoreactivity. The presence of catecholaminergic characteristics in NOS-containing neuronal somata is unusual and raises the question as to their origin. Their heterogeneity suggests different functions for the different subpopulations.


Subject(s)
Dopamine beta-Hydroxylase/metabolism , Kidney/innervation , NADPH Dehydrogenase/metabolism , Neurons/enzymology , Adrenergic Fibers/enzymology , Animals , Fluorescent Antibody Technique , Ganglia, Autonomic/enzymology , Immunoenzyme Techniques , Kidney/cytology , Male , Parasympathetic Fibers, Postganglionic/enzymology , Rats , Rats, Sprague-Dawley
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