ABSTRACT
The concept that the mitral valve of the heart is a passive flap that opens and closes like a barn door has been emphasized for decades by medical and biology professors to their students. But experimental findings, which are outlined in this report, support the theory of an active valve. We hypothesize that the two leaflets of the mitral valve are actively contractile; that physical forces generated in the valve itself may stabilize and add precision to the sum of forces that regulate valve movement. This precision could be of critical significance both in the moments preceding, and during, valve opening and closing. Evidence supporting our active valve hypothesis includes the profuse innervation of motor and sensory nerves that are present in the mitral valves of all animals studied. In addition, multiple contractile cell types have been found in the mitral valve, including cardiac muscle cells, smooth muscle cells, and cardiac valvular interstitial cells. In vitro work in our laboratories using the rat mitral valve shows that not only are the valves capable of contraction and relaxation, but that the contractions and relaxations are nerve-mediated. We theorize that the rich innervation and contractile cells in the mitral valve work together to modulate fine-tuning of valve movements and tone, thereby ensuring the integrity of the valve seal. Other investigators have reported that the mitral valve demonstrates contractile activity and that denervation localized to the mitral valve affects valve competence. The evidence for an active mitral valve presented by these and other experimental studies warrant a reexamination of the validity of the passive valve concept. An accurate and full understanding of the precise movements of the valve leaflets and the mechanisms that regulate these movements is likely to provide the information needed to understand and develop treatments for many different cardiac valve problems, including mitral valve diseases such as prolapse and myxomatous degeneration. In view of the available experimental evidence, the concept that the mitral valve functions only as a passive structure is challenged by numerous anomalies. A reinterpretation of the concept of valve function that incorporates active as well as passive roles for the valve leaflets and other components of the valve apparatus would have significant implications both for the directions taken in research involving the cardiac valves and for the approaches to treatment.
Subject(s)
Mitral Valve/physiology , Animals , Biomarkers , Calcitonin Gene-Related Peptide/metabolism , Heart Atria , Heart Ventricles , Humans , Immunohistochemistry , Microscopy, Confocal , Mitral Valve/chemistry , Mitral Valve/cytology , Mitral Valve/enzymology , Mitral Valve/innervation , Motor Neurons/cytology , Muscle Contraction , Muscle Relaxation , Muscle Tonus , Muscle, Smooth/cytology , Muscle, Smooth/physiology , Myocytes, Cardiac/cytology , Myocytes, Cardiac/physiology , Neurons, Afferent/cytology , Neuropeptide Y/metabolism , Norepinephrine/metabolism , Tyrosine 3-Monooxygenase/metabolismABSTRACT
It presents images (axial plane) and information related to the anatomy of the human body (head, neck, shouders, upper arm, upper and middle torax, upper limb, lower torax, abdomen, pelvis, perinium, hip, upper thigh, and lower limb), topography of the torax and abdomen, and bibliography.
Subject(s)
Head/anatomy & histology , Neck/anatomy & histology , Shoulder/anatomy & histology , Arm/anatomy & histology , Thorax/anatomy & histology , Lung/anatomy & histology , Extremities/anatomy & histology , Abdomen/anatomy & histology , Perineum/anatomy & histology , Hip/anatomy & histology , Thigh/anatomy & histology , Reference Books , AnatomyABSTRACT
Previous reports demonstrated that mammalian atrioventricular (AV) valves possess a dense nerve plexus, consisting of nerve subpopulations which differ from each other in densities and patterns of distribution in the valves, and which may have sensory or motor roles in valve function. Although there is extensive evidence that age-related changes occur in autonomic nerves of animals and humans (Daly et al. J. Pharm. Exp. Ther., 1988;245(3):798-803; Ingall et al. Aust. NZ J. Med., 1990;20:570-577; Tumer et al. Exp. Gerontol., 1992;27:301-307), and that these changes contribute to changes in cardiac function (Klausner and Schwartz Clin. Geriat. Med., 1985;1(1):119-114), there is little information about age-related changes in heart valve innervation. In this study, we used acetylcholinesterase (AChE) histochemistry to localize and compare qualitative and quantitative changes in the innervation of the mitral valves in young adult and aged animals of three species. Young adult and aged guinea pigs, mice, and Wistar and Fischer 344 rats were anesthetized with Nembutal, the hearts removed, and the mitral valves dissected out and processed for AChE localization. Camera lucida drawings of the AChE-positive nerves in representative segments of valve cusps were made directly from slides; these drawings were digitized and subjected to computer-assisted image analysis to obtain quantitative information about nerve plexus density in the valves. All three animal species showed profuse AChE-positive innervation in the mitral valves of young adult animals, and decreases in the density of this innervation in aged animals. The most striking loss of innervation, compared to the young adult, occurred in the mitral valves of aged Fischer 344 rats, in which large regions of the valves appeared virtually devoid of nerves. Further studies are needed to investigate whether and to what extent age-related losses in heart valve innervation affect valvular structure and function.
Subject(s)
Mitral Valve/innervation , Acetylcholinesterase/analysis , Age Factors , Animals , Biomarkers/analysis , Female , Guinea Pigs , Histocytochemistry , Image Processing, Computer-Assisted , Male , Mice , Mice, Inbred CBA , Rats , Rats, Inbred F344 , Rats, WistarABSTRACT
Using confocal fluorescence microscopy we studied, in whole mounts of heart mitral valves of young adult and aged Fischer 344 rats, the distribution of nerves containing the catecholamine marker tyrosine hydroxylase (TH) or the synthetic enzyme marker for nitric oxide, nitric oxide synthase (NOS). TH-IR was localized in two separate nerve plexuses which do not intermingle. The 'major' plexus arose from the annulus region, traversed the basal zone of the valve, and ramified in the intermediate zone to form a dense network of fine fibers. The 'minor' plexus was restricted to the distal zone and originated from bundles that ascended the chordae tendineae to enter the valve cusp. A concentric zone located between the major and minor plexuses was devoid of TH-IR nerve fibers. Both plexuses demonstrated (i) nerves that contained numerous varicosities along the length of each fiber, (ii) many terminal axons and (iii) different shaped terminal axon endings. With age, the density of TH-IR innervation in the mitral valve was markedly reduced; and nerve fibers of the minor plexus were limited to the chordae tendinae, without extending into the valve cusp itself. NOS-IR fibers in the mitral valve formed a loose network that extended from the annulus to more than halfway down the cusp. The varicose beads of the terminal NOS-IR axons appeared to become progressively smaller and less intensely fluorescent until they disappeared at the terminal endings, which showed no specializations. No NOS-IR fibers were observed in the distal zone of the valve leaflet or in the chordae. In the aged mitral valve, the density of NOS-IR nerves was decreased, as compared with NOS-IR innervation in the young adult valve. The existence of TH and NOS as well as other signal molecule markers in heart valve nerves and the disparate patterns of their distribution and localization provide evidence supporting the theory that heart valve nerves form a complex reflexogenic control system in the mitral heart valve. In summary, two distinct neural architectures are described for TH-IR and NOS-IR valve nerves, respectively. The former are believed to be axons dedicated to sympathetic motor functions. The NOS-IR valve nerves may have sensory and/or postganglionic parasympathetic motor functions. An implication of these findings is that different, but perhaps related, valve functions may be mediated by separate, dedicated circuits.
Subject(s)
Adrenergic Fibers/immunology , Aging/physiology , Mitral Valve/innervation , Nitric Oxide Synthase/analysis , Tyrosine 3-Monooxygenase/analysis , Adrenergic Fibers/chemistry , Animals , Antibody Specificity , Immunohistochemistry , Male , Microscopy, Confocal , Nitric Oxide Synthase/immunology , Rats , Rats, Inbred F344 , Tyrosine 3-Monooxygenase/immunologyABSTRACT
The objective of this study was to determine whether and to what extent age-related changes occur in atrioventricular (AV) heart valve innervation. The AV valves from three young adult (3 months) and three older (> 24 months) female guinea pigs were studied. An acetylcholinesterase (AChE) localization method was used to prepare valve whole mounts for analysis. Two methods were used to assess nerve fiber density. Segments of the valves were drawn using a camera lucida/Nikon optiophot system. The density of nerve fibers was calculated from digitized images. The density of nerve fibers was also calculated by counting the points at which the nerve plexus intersected with the grid lines of an ocular graticule. In the bicuspid and tricuspid valves of the older guinea pigs, we observed a marked diminution in the densities of the nerve plexus, particularly in the basal zone, towards the free edges of the valve cusps, and in the chordae tendineae. Whole mount preparations such as those used in our morphological studies of the AV innervation may assist in elucidating the changes in other autonomic nerve plexuses with aging. Further work is required to establish whether and to what extent the loss of valve innervation influences the effectiveness of closure of the valves.
Subject(s)
Acetylcholinesterase/analysis , Heart Valves/innervation , Nerve Fibers/enzymology , Age Factors , Animals , Female , Guinea Pigs , Staining and LabelingABSTRACT
The specific functions of the numerous substance P (SP) nerve fibers present within the gastrointestinal tract are not clearly defined. This study examines both functional aspects and distribution of immunoreactive SP (IR-SP) in the canine gastroesophageal junctional (GEJ) region. Lower esophageal sphincter pressure (LESP), mean arterial pressure (MAP), pulse rate (PR), and respiratory rate (RR) were monitored before and after topical application of 2 ml capsaicin (8-methyl-N-vanillyl-6-nonenamide) to the distal esophageal mucosa of anesthetized dogs. Animals then underwent a capsaicin desensitization protocol over a 12-day period. The responses of monitored variables were compared on Day 1 and Day 12 of repetitive capsaicin application. Immunohistochemistry and radioimmunoassay (RIA) were performed on GEJ segments to study the distribution and content of IR-SP in both control (untreated) and capsaicin-treated dogs. The IR-SP was extracted from tissue for RIA and analysis by reverse-phase high-performance liquid chromatography (HPLC). On Day 1, a 2-ml capsaicin application stimulated increases in LESP (44.3 +/- 7.8 cm H2O; P < 0.05), MAP (48 +/- 8.7 mm Hg; P < 0.05), PR (52.6 +/- 20.5 beats/min; P < 0.05), and RR (26.3 +/- 15.6 breaths/min; P > 0.2). No response was observed on Day 12 of treatment. This was accompanied by a 43.3% decrease of IR-SP content in the mucosa of the distal esophagus of desensitized animals. Capsaicin applied at greater concentrations on Day 12 stimulated a return of responses (P < 0.05). Ganglia, cell bodies, nerve fascicles, and neurites stained positively for IR-SP. IR-SP content was markedly higher in esophageal mucosa than in gastric mucosa (P < 0.05). The authenticity of the IR-SP molecule was confirmed by elution time on HPLC. In conclusion, repetitive capsaicin application induced a state of homologous desensitization which was accompanied by a partial depletion of mucosal SP. The GEJ region contains a high SP content with a broad neural distribution. These findings are consistent with the hypothesis that SP may act as a neurotransmitter for chemonociceptive stimuli in the canine distal esophagus.
Subject(s)
Esophagus/physiology , Stomach/physiology , Substance P/physiology , Animals , Blood Pressure/drug effects , Capsaicin/pharmacology , Chromatography, High Pressure Liquid , Dogs , Esophagus/chemistry , Esophagus/innervation , Immunohistochemistry , Mucous Membrane/drug effects , Mucous Membrane/physiology , Neurons/chemistry , Pressure , Pulse/drug effects , Radioimmunoassay , Respiration/drug effects , Stomach/chemistry , Stomach/innervation , Substance P/analysisABSTRACT
BACKGROUND: Neuroplastic changes following ileum hypertrophy have been reported in intrinsic enteric neurons. The hypothesis in the present study was that intestinal hypertrophy induces neuronal changes in dorsal root ganglia (DRG). METHODS: Under sodium pentobarbital anesthesia, partial obstruction was produced in the rat by tying a plastic ring around the terminal loop of ileum. Fast Blue (FB) (Sigma, St. Louis, MO) was injected into the obstructed ileum wall, and the rat was perfused after 8 days. DRG were immunostained and examined to identify and measure sizes of perikarya containing FB and/or calcitonin gene-related peptide (CGRP) or FB and/or substance P (SP). RESULTS: Of the DRG neurons that projected to the ileum in control or obstructed animals, approximately 50% were CGRP-immunoreactive (IR) and 30% were SP-IR (colchicine pretreatment was not used). Neurons that projected to the obstructed ileum were increased in size compared with neurons in nonobstructed controls. Some of these neurons were CGRP-IR or SP-IR; some were large FB-labeled neurons that were not SP-IR or CGRP-IR. CONCLUSIONS: The morphology of sensory autonomic neurons in adult animals is influenced by dynamic interactions with the targets they innervate, whether directly or transneuronally.
Subject(s)
Ganglia, Spinal/pathology , Intestinal Obstruction/pathology , Neurons, Afferent/pathology , Animals , Calcitonin Gene-Related Peptide/analysis , Calcitonin Gene-Related Peptide/immunology , Hypertrophy , Male , Nerve Growth Factors/analysis , Neurons, Afferent/chemistry , Rats , Rats, Sprague-Dawley , Substance P/analysis , Substance P/immunologyABSTRACT
Schistosomiasis mansoni is a parasitic disease in which granulomas form around schistosome eggs in the liver and intestines. The purpose of this study was to determine the alterations in the intrinsic innervation of the distal ileum and proximal colon resulting from schistosomiasis. Using murine schistosomiasis mansoni, we examined light microscopic preparations stained with osmium-zinc iodide or the dihydronicotinamide adenine dinucleotide: nitro BT oxidoreductase (NADH) method. We also examined specific populations of peptidergic nerves (vasoactive intestinal polypeptide and substance P) using an avidin-biotin complex (ABC) immunohistochemical technique. We found that granulomas focally destroyed the enteric nerves. Occasionally nerves were found within granulomas, particularly at the periphery of the lesions. Nerve cell bodies close to granulomas had altered staining, which included increased staining for vasoactive intestinal polypeptide. The distribution of nerve injury varied between the 2 enteric segments studied. In the distal ileum, the principal injury was to the myenteric plexus; whereas, the submucous and mucosal plexuses were predominantly damaged in the proximal colon. The physiologic significance of this injury to the enteric nerves requires elucidation.
Subject(s)
Colon/innervation , Granuloma/pathology , Ileum/innervation , Schistosomiasis mansoni/pathology , Animals , Female , Immunohistochemistry , Mice , Mice, Inbred CBA , Substance P/analysis , Vasoactive Intestinal Peptide/analysisABSTRACT
Lower esophageal sphincter (LES) effects produced by the mammalian tachykinins were evaluated in anesthetized dogs. The distribution and content of substance P (SP) and neurokinin A (NKA) in the region of the canine gastroesophageal junction was also studied. SP and NKA stimulated a linear dose-dependent contraction of the LES after intra-arterial administration. Neurokinin B (NKB) failed to stimulate an increase in LES pressure (LESP). SP was characterized by an immediate but short-lived contraction followed by a period of relaxation. NKA stimulated a potent LES contraction that was slow in onset but long-lasting. On an equimolar basis, both SP and NKA were approximately 100 times more potent LES stimulants than bethanechol or phenylephrine. Pretreatment with atropine (muscarinic blockade) or tetrodotoxin (neural blockade) inhibited the effect produced by SP. NKA appeared to stimulate LES contraction independent of neural or cholinergic mechanisms. Radioimmunoassay revealed a regional variation in tachykinin content in the gastroesophageal junction. Ganglia, cell bodies, nerve fascicles, and neurites stained specifically for both SP and NKA. The variable effects, potencies, and mechanisms of action observed in this study suggest the presence of specific tachykinin receptor subtypes in the gastroesophageal junction. Both SP and NKA were found to have a broad neural distribution in this region. These findings suggest that the tachykinins may play an important role in neuroregulation of LES smooth muscle.
Subject(s)
Esophagogastric Junction/physiology , Tachykinins/physiology , Animals , Atropine/pharmacology , Chromatography, High Pressure Liquid , Dogs , Dose-Response Relationship, Drug , Esophagogastric Junction/drug effects , Esophagogastric Junction/innervation , Esophagogastric Junction/metabolism , Esophagus/metabolism , Gastric Mucosa/metabolism , Immunohistochemistry , Manometry , Muscle Contraction/drug effects , Neurokinin A/metabolism , Neurokinin A/pharmacology , Neurokinin A/physiology , Neurokinin B/metabolism , Neurokinin B/pharmacology , Neurokinin B/physiology , Neurons/metabolism , Phentolamine/pharmacology , Radioimmunoassay , Substance P/metabolism , Substance P/pharmacology , Substance P/physiology , Tachykinins/metabolism , Tetrodotoxin/pharmacologyABSTRACT
In this series of studies, the innervation patterns of whole-mount preparations of bicuspid and tricuspid valves were studied by light microscopy in the mouse, rat, guinea pig, and opossum. The acetylcholinesterase-positive networks of nerve fibers showed many similarities in the basic patterns of valve innervation in all of the species studied, but several interspecies variations were observed. The basal zone of the valve adjacent to the fibromuscular atrioventricular ring displayed the most dense plexus of nerves, with acetylcholinesterase-positive fibers being seen across the width of the valve. In the intermediate zone of the valve, less dense plexuses of nerve fibers were found; and these were more numerous in the cuspal areas and less numerous in the intervening commissural areas. In the distal portions of the valve, nerve networks arborized extensively, with some of their nerve fibers extending toward the chordae tendineae and the free edges of the valve cusps. Only in the guinea pig and opossum did these fibers reach the free margin of the valve cusp, where they either ended directly as free nerve endings or lay parallel to the free edge of the cusp, often running between adjacent chordae tendineae. Although the patterns of innervation were similar in both bicuspid and tricuspid valves, the innervation density of the bicuspid valve was greater than that of the tricuspid valve for each species examined. A distinguishing feature of guinea pig and opossum tricuspid valves was that their chordae tendineae were relatively more prominent and more densely innervated than the bicuspid chordae tendineae. Free nerve endings with no light microscopic evidence of specialization were present throughout the bicuspid and tricuspid valves of all species studied. Some nerve endings in the opossum showed evidence of specialization, with brush-like arborizations leading to presumed free terminals seen chiefly in the distal zone of the valve cusps. Although some general tendencies were apparent, we have demonstrated that interspecies heterogeneity exists in the terminal networks of the atrioventricular valves of mouse, rat, guinea pig, and opossum.
Subject(s)
Mitral Valve/innervation , Opossums/anatomy & histology , Rodentia/anatomy & histology , Tricuspid Valve/innervation , Animals , Female , Guinea Pigs , Mice , Mice, Inbred CBA , Rats , Rats, Inbred Strains , Species SpecificityABSTRACT
These experiments utilize a paradigm developed to study plastic responses of peptidergic neurons in a discrete brain area following deafferentation. The central nucleus of the amygdala (CNA) is richly innervated by somatostatin-immunoreactive (SS-I) terminal axons. In the course of preliminary light microscopic (LM) investigations by this laboratory, changes were observed in the density of presumed SS-I terminals in the rat CNA after lesioning the medial input. The LM finding of increased density of presumed SS-I terminals in the CNA at the 10-day post-lesion stage underscored the need for a quantitative electron microscopic (EM) study of the SS-I components, including an evaluation of synaptic events at different survival periods. At the 3-day post-lesion stage, EM examination showed degenerating axons in the lesioned CNA, many already engulfed by astrocytes. None of the degenerating profiles were SS-I, supporting the view that the lesion did not interrupt, to any significant extent, SS-I axons entering the nucleus. EM surveys of the 10-day post-lesion material demonstrated that degenerated profiles had almost completely disappeared. Numbers of SS-I axon terminals, particularly of smaller-sized profiles, were increased by 22% over control value. Synaptic frequency was decreased by 16% below control value. Numbers of SS-I terminals making synapses were increased 3.4% above control value. At the 30-day post-lesion stage, the total number of SS-I terminal axons had increased 86% over controls, whereas the synaptic frequency had decreased by about a third below controls. The absolute number of SS-I terminals engaging in synapses had increased by 24% over controls. The 90-day post-lesion CNA showed a further increase in the number of SS-I axon profiles: 136% over control value. The synapse-to-axon ratio (synaptic frequency) of 27% was similar to that observed for the CNA from the unlesioned side or from unoperated animals. At this stage the number of SS-I synapses had increased by 135% over controls. This model presents many possibilities for studying neuroplasticity, particularly involving peptidergic neurons of the central autonomic nervous system.
Subject(s)
Amygdala/physiology , Axons/physiology , Neuronal Plasticity , Somatostatin/metabolism , Amygdala/ultrastructure , Animals , Axons/ultrastructure , Immunohistochemistry , Male , Microscopy, Electron , Rats , Rats, Inbred Strains , Synapses/physiology , Synapses/ultrastructureABSTRACT
We localized immunoreactive substance P and measured its content throughout the opossum esophagus. Substance P-like immunoreactive nerve fibers were more abundant in muscularis mucosae than in the longitudinal smooth muscle layer and more abundant in the latter than in the circular smooth muscle layer. The distribution of substance P-like immunoreactive nerve fibers in the circular muscle layer of the esophagogastric junction was comparable to that of the esophageal body. Immunoreactive fibers were also found on and in arteries, submucosal glands, and epithelium, but none were seen in striated muscle. Both the myenteric and submucous plexuses contained substance P-like immunoreactive nerve cell bodies and processes. In the muscularis propria, the content of substance P-like immunoreactive peptide (pg/mg protein, means +/- 1 SE) in the smooth muscle region of the esophageal body (5.9 +/- 0.6) exceeded that in the striated muscle region (2.5 +/- 0.2) and at the esophagogastric junction (1.8 +/- 0.5), but the latter two values were similar (P less than 0.05). Mucosal content of substance P in the region of the esophagogastric junction (1.2 +/- 0.1) differed from that of the smooth muscle region of the esophageal body (9.2 +/- 2.6) but not (P greater than 0.05) from that of the striated-muscle region (5.9 +/- 1.0). The broad distribution and diversity of immunoreactive structures suggest that substance P may have both sensory and motor functions in the esophagus.
Subject(s)
Esophagus/metabolism , Opossums/metabolism , Substance P/metabolism , Animals , Deglutition , Female , Immunohistochemistry , Male , Muscle, Smooth/metabolism , Nerve Fibers/metabolism , Substance P/physiologyABSTRACT
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.
Subject(s)
Ileum/innervation , Neuronal Plasticity/physiology , Neurons/physiology , Animals , Ileum/metabolism , Ileum/physiology , Immunohistochemistry , Male , Microscopy, Electron , Models, Biological , Myenteric Plexus/metabolism , Myenteric Plexus/physiology , Myenteric Plexus/ultrastructure , Neurons/metabolism , Neurons/ultrastructure , Rats , Rats, Inbred Strains , Substance P/metabolism , Vasoactive Intestinal Peptide/metabolismABSTRACT
Neurotensin (NT) was demonstrated in the central nucleus of the rat amygdala (CNA) using a modification of the avidin-biotin complex immunohistochemical technique. Electron-dense reaction product (particles were 15-25 nm in diameter) was localized in perikarya, dendrites, axons, and axon terminals. It was found also associated with profiles of rough endoplasmic reticulum, mitochondria, microtubules, and small agranular as well as large granular vesicles. In distal dendrites, the reaction product was associated with microtubules, vesicles, and postsynaptic densities. Axon terminals of three types formed synaptic contracts with NT-immunoreactive neurons in the CNA: one was characterized by numerous round or oval agranular vesicles, the second by numerous pleomorphic vesicles, and the third by agranular vesicles that were loosely distributed and pleomorphic. All three types formed symmetric axosomatic and asymmetric axodendritic contacts. NT-immunoreactive axon terminals containing small round agranular vesicles stood out clearly from the intermingling profiles of immunonegative structures. We found numerous glomeruli, each consisting of a central NT-immunoreactive dendrite surrounded by all three types of axon terminals. We observed that some NT-immunoreactive terminals formed symmetric axoaxonal contacts with each other, providing evidence for the presence of local NT-to-NT circuits, whereas many others synapsed with axon terminals devoid of NT immunoreactivity.
Subject(s)
Amygdala/ultrastructure , Neurotensin/analysis , Amygdala/cytology , Animals , Axons/ultrastructure , Dendrites/ultrastructure , Immunohistochemistry , Male , Microscopy, Electron , Neurotensin/immunology , Rats , Rats, Inbred Strains , Synapses/ultrastructureABSTRACT
Physiologic evidence that vasoactive intestinal polypeptide (VIP) regulates esophageal smooth muscle in the opossum has been gathered without knowledge of the distribution of VIP in that organ. We examined planar sections stained for VIP by the avidin-biotin complex method, measured VIP content in mucosa and muscularis propria by radioimmunoassay, and compared neural structures reactive to VIP antiserum with those revealed by osmication in the presence of zinc iodide. Immunoreactive terminal nerves interlaced smooth muscle bundles in all layers in all smooth muscle regions, formed loose tangled knots about widely dispersed muscle cells in striated muscle, and supplied vessels and submucosal glands. Bipolar interstitial cells in the circular muscle layer stained by osmication were not VIP-immunoreactive. Perikarya in both submucous and myenteric plexuses were VIP-immunoreactive. Vasoactive intestinal polypeptide-immunoreactive oval cells with round unstained nuclei and a faintly granular cytoplasm were scattered in the muscle in all regions and were concentrated in the planes of the plexuses. Vasoactive intestinal polypeptide content of muscularis propria in the smooth muscle esophageal body exceeded (p less than or equal to 0.05) that in the striated muscle esophageal body and the sphincter region, but contents in the latter two regions did not differ (p greater than or equal to 0.05). Mucosal content exceeded that of muscularis propria. The broad distribution and diversity of immunoreactive structures suggest that VIP may have functions in this organ besides the regulation of smooth muscle.
Subject(s)
Muscle, Smooth/metabolism , Opossums/metabolism , Vasoactive Intestinal Peptide/metabolism , Animals , Esophagus/immunology , Esophagus/metabolism , Female , Fluorescent Antibody Technique , Histocytochemistry , Immunoenzyme Techniques , Male , Muscle, Smooth/immunology , Staining and Labeling/methodsABSTRACT
Catecholaminergic (CA) nerve terminals in the paraventricular hypothalamic nucleus (PVN) of adult rats were studied at 4, 21, 56 and 180 days after a single injection of 6-hydroxydopamine (6-OHDA) neurotoxin into the right lateral ventricle of the brain. We previously described and quantified the extent of CA terminal sprouting in the PVN after 6-OHDA lesions. For this communication we studied parameters, specifically the bouton sizes and the synaptic frequencies of CA terminals during the renewal process, and evaluated how changes of these parameters are related to axonal sprouting. The CA boutons were identifiable in the electron microscope by exhibiting small granular vesicles (SGVs) after central administration of 5-hydroxydopamine (5-OHDA) marker. The marked CA boutons were measured and further categorized according to whether or not they were associated with distinct synaptic specializations at various post-lesion stages. The average sizes of CA boutons were strikingly similar in their diameters (1.0 micron) for both control and experimental tissues. However, CA boutons larger than 2.1 micron were rare and seen more often in the experimental tissues with 6-OHDA lesion and were sustained up to 180 days after lesions. Catecholaminergic profiles with ultrastructural features of growth cones were also seen in the PVN following the 6-OHDA lesions, indicating that there is growth activity in the PVN after 6-OHDA lesion. There were 33% of CA boutons in the PVN from the control tissues that appeared to have synaptic contacts.(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Catecholamines/physiology , Hydroxydopamines/pharmacology , Neuronal Plasticity , Paraventricular Hypothalamic Nucleus/physiology , Adrenergic Fibers/physiology , Animals , Male , Microscopy, Electron , Oxidopamine , Paraventricular Hypothalamic Nucleus/drug effects , Paraventricular Hypothalamic Nucleus/ultrastructure , Rats , Rats, Inbred Strains , Synapses/ultrastructure , Synaptic TransmissionABSTRACT
Amine-containing small intensely fluorescent (SIF) cells are ubiquitous in vertebrate sympathetic ganglia and, in some species, SIF cells have been identified as interneurons. The hypothesis proposed in this study is that SIF cells in superior sympathetic ganglia of the guinea pig function as interneurons, with efferent connections characteristic for the species. Fluorescence (catecholamine) microscopy and 5-hydroxydopamine marker for electron microscopy were used to study SIF cells, their processes and connections in this ganglion. Brightly fluorescent fibers were seen attached to virtually all SIF cells, and were of two types. The first type, single or arranged in cords, interconnected elements of the SIF-cell system; these apparent linkages joined individual SIF cells as well as adjacent clusters. The electron-microscopic evidence for synaptic contacts between SIF cells warrants the claim that integrated action is a presumed function of these elements. The second type of SIF-cell process was generally of greater length. These individual, branching fibers made presumed connections with dendrites of most principal ganglionic neurons. This arrangement suggested by histofluorescence preparations was confirmed by electron microscopy to involve synaptic connections, and the postsynaptic element was shown to be continuous with the perikaryon of the principal ganglionic neuron. Ultrastructural evidence that collections of dense-cored vesicles occur within processes of both principal ganglionic neurons and SIF cells, in proximity to unsheathed portions of plasma membrane, leads to the conclusion that interstitial diffusion of catecholamine from both may occur; the finding of SIF cell processes adjacent to fenestrated blood vessels suggests that catecholamine may also be transported through capillaries.
Subject(s)
Ganglia, Sympathetic/cytology , Animals , Ganglia, Sympathetic/drug effects , Ganglia, Sympathetic/ultrastructure , Guinea Pigs , Hydroxydopamines/pharmacology , Microscopy, Electron , Microscopy, Fluorescence , Neurons/cytology , Neurons/ultrastructureABSTRACT
Glass microknife lesions and immunocytochemistry were used to evaluate luteinizing hormone-releasing hormone (LH-RH)- and somatostatin (SS)-immunoreactive pathways from the preoptic region to the rat median eminence. Cuts were so placed that axons of more caudally located neurons in the periventricular hypothalamic areas were spared. Light and EM observations of LH-RH-immunostained preparations indicated that following the midline periventricular cuts the density of LH-RH labelled axons and axon terminals in the ME appeared similar to that of nonlesioned animals. Following bilateral lateral hypothalamic cuts placed between the preoptic area and the ME, LH-RH immunostaining in the ME was markedly reduced. This provides evidence that the preponderance of LH-RH axons originating from the preoptic area reach the ME by a lateral hypothalamic route. In contrast to the LH-RH findings, midline lesions made using the same coordinates caused a noticeable reduction in SS immunostaining in the accurate nucleus and ME. There was either no change or only minimal change after the lateral cut. Somatostatin axons arising from the preoptic periventricular nucleus take a periventricular route and contribute to median eminence innervation, but much less extensively than the more caudally located somatostatin neurons in the hypothalamic periventricular nucleus [19].
Subject(s)
Gonadotropin-Releasing Hormone/analysis , Median Eminence/analysis , Somatostatin/analysis , Animals , Axons/analysis , Female , Histocytochemistry , Immunochemistry , Microscopy, Electron , Nerve Fibers/analysis , Rats , Rats, Inbred StrainsABSTRACT
Capsaicin was given subcutaneously to guinea pigs and the effect on substance P-immunoreactive (SP-I) fibers in the celiac/superior mesenteric and inferior mesenteric ganglia was observed at 2 day and 8-10 day intervals. Capsaicin (125 mg) treatment led to almost total disappearance of SP-I fibers from all areas examined in both short- and long-term animals. This effect applied equally to the dense network of varicose SP-I fibers and to basket-like SP-I contacts with principal ganglionic neurons. The effect of capsaicin on SP-I fibers in the mesenteric ganglia provides a strong indication that these fibers represent a homogeneous population of visceral sensory afferents. This is supported by other lines of anatomical evidence in the literature. Taken together with studies that have shown axodendritic contact of SP-I terminals on principal ganglionic neurons and neuro-modulatory effects of SP on these neurons, it may be hypothesized that SP-I fibers in the mesenteric ganglia represent collaterals of visceral sensory afferents forming a subspinal feedback arc.