Morphometry and acetylcholinesterase activity of the myenteric plexus of the wild mouse Calomys callosus

The myenteric plexus of the digestive tract of the wild mouse Calomys callosus was examined using a histochemical method that selectively stains nerve cells, and the acetylcholinesterase (AChE) histochemical technique in whole-mount preparations. Neuronal density was 1,500 +/- 116 neurons/cm2 (mean +/- SEM) in the esophagus, 8,900 +/- 1,518 in the stomach, 9,000 +/- 711 in the jejunum and 13,100 +/- 2,089 in the colon. The difference in neuronal density between the esophagus and other regions was statistically significant. The neuron profile area ranged from 45 to 1,100 microns2. The difference in nerve cell size between the jejunum and other regions was statistically significant. AChE-positive nerve fibers were distributed within the myenteric plexus which is formed by a primary meshwork of large nerve bundles and a secondary meshwork of finer nerve bundles. Most of the nerve cells displayed AChE activity in the cytoplasm of different reaction intensities. These results are important in order to understand the changes occurring in the myenteric plexus in experimental Chagas' disease.     

Distribution of nitric oxide synthase in the esophagus of the cat and monkey

The distribution of nitrergic neurons and processes in the esophagus of the cat and monkey was studied by light microscopic immunocytochemistry using a specific antibody against purified rat brain nitric oxide synthase and immunoperoxidase procedures. Immunoreactive nerve fibers were found pervading the myenteric plexus, submucous plexus and plexus of the muscularis mucosae, and particularly in the lower esophagus a few immunoreactive fibers entered the epithelium as free nerve endings, some of which derived from perivascular fibers. In the upper esophagus immunoreactive motor end-plates were found in the striated muscle. Thirty-forty-five percent of neuronal cell bodies found in the intramural ganglia and along the course of nerve fiber bundles were immunoreactive and were of the three morphological types earlier described. In the intramural ganglia immunoreactive nerve fibers formed a plexus in which varicose nerve terminals were in close relation to immunoreactive and non-immunoreactive neurons. The intramural blood vessels that crossed the different layers of the esophageal wall were surrounded by paravascular and perivascular plexuses containing immunoreactive nerve fibers. The anatomical findings suggest that nitric oxide is involved in neural communication and in the control of peristalsis and vascular tone in the esophagus. In the lower esophagus a few nitrergic nerve fibers are anatomically disposed to subserve a sensory-motor function.     

Oxygen sensing by the global regulator, FNR: the role of the iron-sulfur cluster

Pituitary adenylate cyclase-activating peptide (PACAP)-immunoreactive (IR) neurons in the myenteric and submucosal plexus of the rat small and large intestine were examined by immunostaining with purified polyclonal antiserum against PACAP (1-15), using both light and electron microscopy. Many PACAP-IR neuronal cell bodies and fibers were found in the myenteric and submucosal plexus. Many of the PACAP-IR fibers originated from the cell bodies of the myenteric and submucosal ganglia. The ganglia were also innervated by PACAP-IR fibers. PACAP-IR fibers penetrated both the circular and longitudinal muscle layers, confirming the previous observations indicating that PACAP neurons act as motor neurons. Ultrastructural study demonstrated that PACAP-IR nerve terminals formed synaptic contacts with PACAP-IR nerve cell bodies or dendritic processes. This observation suggests that PACAP-IR neurons innervate other PACAP-IR neurons, and that PACAP neurons work as interneurons in the enteric nervous system. PACAP-IR nerve cells received not only PACAP-positive nerve terminal input also PACAP-negative nerve terminal input. It also suggests that PACAP neurons are regulated not only by PACAP-IR enteric neurons, but also by neurons originating elsewhere. Our observations support the view that PACAP-IR neurons are involved in the control of gut motility.     

Projections of pelvic autonomic neurons within the lower bowel of the male rat: an anterograde labelling study

The tissues of the large intestine which receive an innervation by neurons of the major pelvic ganglia were identified following in vivo and in vitro anterograde labelling with the lipophilic tracer 1,1'didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate in the male rat. The primary target in the gut of major pelvic ganglion neurons is the myenteric plexus of the distal colon and the rectum. The serosal ganglia, on the surface of the most distal region of the rectum and the circular muscle of the distal colon and rectum were less densely innervated. The pelvic ganglia do not innervate the longitudinal muscle, submucosal blood vessels, submucosal plexus, or mucosa. The pelvic supply reaches the bowel via two groups of rectal nerves and branches of the penile nerves. All of these connections also carry the axons of viscerofugal neurons from the bowel, some of which have terminal axons in the major pelvic ganglia. Finally, the different nerves supplied different targets. In particular, while the rectal nerves carried pelvic axons supplying the myenteric plexus, circular muscle, and serosal ganglia, the penile nerves only innervated the serosal ganglia. In addition, the two groups of rectal nerves innervated slightly different regions of the bowel and provided different projection patterns. However, successful in vivo labelling was achieved in only 6/12 animals and while all in vitro experiments resulted in successful labelling, it was clear that only a proportion of pelvic projections in any given nerve were labelled. These studies have shown that the major pelvic ganglia are primarily involved in the control of motility, but not of vascular and secretomotor functions. Thus pelvic neurons do not innervate the same range of target tissues within the bowel as the prevertebral ganglia. This study has also shown that the different pathways to the gut from the major pelvic ganglia innervate different tissues, suggesting that the autonomic innervation of the gut is not homogeneous along its length.     

Morphology and histochemistry of the rabbit pancreatic innervation

Stimulation of extrinsic nerves markedly alters pancreatic endocrine and exocrine secretion, yet little is known of the neurochemical organization and physiologic roles of specific neural pathways within the pancreas. Here we report histochemical staining for acetylcholinesterase (AChE), NADPH-diaphorase (NADPH-d), nitric oxide synthase (NOS), and several neuropeptides to identify the neurotransmitter content of rabbit pancreatic nerves. An extensive network of AChE-positive nerve fibers was found throughout the islets, acini, ducts, ganglia, and blood vessels. All pancreatic neurons were AChE positive, two thirds were NADPH-d positive, and many were NOS positive. Ganglia in the head/neck region were connected to the duodenal myenteric plexus by AChE- and NADPH-d-positive fibers, and NADPH-d-positive pancreatic neurons appeared to send processes toward both the duodenum and pancreas. Many pancreatic neurons were vasoactive intestinal peptide (VIP) positive, and VIP nerve terminals were abundant in ganglia, acini, islets, and ducts. Pituitary adenylate cyclase-activating peptide (PACAP-38)-positive fibers also were observed within acini and passing through ganglia. Substance P (SP)-, calcitonin gene-related peptide (CGRP)-, and dopamine beta-hydroxylase (DBH)-positive fibers were abundant along blood vessels and ducts, and varicose fibers were observed in pancreatic ganglia. Fine galanin-positive fibers were also occasionally observed running with blood vessels and through ganglia. Thus the rabbit pancreas receives a dense, diverse innervation by cholinergic, adrenergic, and peptidergic nerves and cholinergic pancreatic neurons, most also containing VIP or NOS or both, appear to innervate both endocrine and exocrine tissue, and may mediate local communication between the duodenum and pancreas.     

The presence of aromatic L-amino acid decarboxylase in certain intestinal nerve cells

The presence of aromatic 1-amino acid decarboxylase (AADC) in nerve cell bodies of the intrinsic plexuses of the guinea-pig small intestine was demonstrated by incubating segments of intestine with 1-dopa in the presence of an inhibitor of monoamine oxidase, pargyline. After such incubation, some nerve cell bodies gave a fluorescence histochemical reaction indicative of the presence of a decarboxylated product of 1-dopa, probably dopamine. No fluorescence reaction occurred in the unincubated control or if the inhibitor of AADC, RO 4-4602, was included in the incubation mixture. The AADC-containing cell bodies apparently do not take up and store dopamine, because no fluorescence could be detected after incubation with dopamine and a monoamine oxidase inhibitor. The AADC-containing cells were found in about half of the ganglia of the submucous plexus of the guinea-pig small intestine, but were considerably less frequent in the myenteric plexus. They were also found in the other areas examined in this study, that is, in both enteric plexuses of the guinea-pig distal colon and of the small intestines of rabbits and rats.     

Morphological and immunohistochemical identification of neurons and their targets in the guinea-pig duodenum

Nerve circuits within the proximal duodenum were investigated using a combination of immunohistochemistry for individual neuron markers and lesion of intrinsic nerve pathways to determine axon projections. Cell shapes and axonal projections were also studied in cells that had been injected with a marker substance. Several major neuron populations were identified. Calbindin immunoreactivity occurred in a population of myenteric nerve cells with Dogiel type II morphology. These had axons that projected to other myenteric ganglia, to the circular muscle and to the mucosa. All were immunoreactive for the synthesizing enzyme for acetylcholine, choline acetyltransferase, and some were also immunoreactive for calretinin. Myenteric neurons with nitric oxide synthase immunoreactivity projected anally to the circular muscle. These were also immunoreactive for vasoactive intestinal peptide, and proportions of them had enkephalin and/or neuropeptide Y immunoreactivity. It is suggested that they are inhibitory motor neurons to the circular muscle. A very few (about 2%) of nitric oxide synthase-immunoreactive neurons had choline acetyltransferase immunoreactivity. Tachykinin (substance P)-immunoreactive nerve cells were numerous in the myenteric plexus. Some of these projected orally to the circular muscle and are concluded to be excitatory motor neurons. Others projected to the tertiary plexus which innervates the longitudinal muscle and others provided terminals in the myenteric plexus. Two groups of descending interneurons were identified, one with somatostatin immunoreactivity and one with vasoactive intestinal peptide immunoreactivity. The two most common nerve cells in submucous ganglia were neuropeptide Y- and vasoactive intestinal peptide-immunoreactive nerve cells. Both provided innervation of the mucosa. There was also a population of calretinin-immunoreactive submucous neurons that innervated the mucosal glands, but not the villi. Comparison with the ileum reveals similarities in the chemistries and projections of neurons. Differences include the almost complete absence of nitric oxide synthase immunoreactivity from vasoactive intestinal peptide-immunoreactive interneurons in the duodenum, the projection of calbindin-immunoreactive Dogiel type II neurons to the circular muscle and the absence of tachykinin-immunoreactivity from these neurons.     

Dysplastic nitrergic neurons in the rectum of a patient with rectal ectasia

We present a histochemical study of resected colon from a 13-year-old boy diagnosed with rectal ectasia. Laminar preparations and sectioned tissue of rectum were assayed histochemically for nitric oxide (NO) synthase activity by reducing nitro blue tetrazolium salt in the presence of the cofactor NADPH. Tissue preparations displayed intensely labelled neurons and fibers throughout the gut wall. Laminar preparations of Meissner's plexus showed a hyperplasia of ganglia and NO-related neurons throughout the length of the resected rectum compared with normal bowel. Sectioned tissue of the Auerbach's plexus demonstrated a normal number of ganglia and NO-related neurons. As well, the ectatic bowel showed a proliferation of nerve fibers in keeping with the degree of circular smooth muscle hypertrophy. This proliferation may represent a reactive phenomenon secondary to the functional obstruction. The NO histochemical technique may form the basis of further investigations in defining the cause of this functional obstruction.     

Mechanical and thermal allodynia in chronic central pain following spinal cord injury

The interstitial cells of Cajal (ICC) are found in a number of different locations in the gastrointestinal tract, where they form close associations with both muscle cells and nerve terminals. In this study we examined the embryological origin of ICC in the mouse intestine to determine whether they arise from the neural crest or from the intestinal wall. Segments of intestine were removed from embryonic mice either before or after the arrival of neural crest cells (the precursors of enteric neurons and glial cells) and transplanted under the renal capsule of host (adult) mice and allowed to develop for 18-41 days. In the mouse intestine, antibodies to c-kit protein selectively label ICC at a variety of locations, and antibodies to the NK1 receptor (the receptor for substance P) labels ICC at the level of the deep muscular plexus in the small intestine and a subpopulation of enteric neurons in the large intestine. The presence of neurons in the explants was examined using antisera to neuron-specific enolase, substance P, and calretinin. In segments of small and large intestine explanted after the arrival of neural crest cells, immunoreactive neurons and c-kit- and NK1-immunoreactive ICC were present with a distribution similar to that seen in control tissue at a similar developmental age. In segments of large intestine explanted before the arrival of neural crest cells, neurons were not present; however, c-kit-immunoreactive ICC were present in these aneuronal explants, indicating that ICC do not arise from the neural crest. The source of ICC in mammals is therefore likely to be the mesenchyme of the gut.     

Involvement of the pelvic plexus and the suprarenal ganglia in the neuropeptide Y (NPY) innervation of the cervix and the uterus of the rat

The involvement of the pelvic plexus and suprarenal ganglia in the neuropeptide Y (NPY) innervation of the genital tract was studied in the female rat by means of denervation experiments and retrograde tracing studies. Removal of the paracervical ganglia caused a significant decrease of the NPY-immunoreactive nerve density and NPY concentration in the lower part of the genital tract: cervix, uterine body and lower part of the uterine horn. The decrease in NPY concentration in these three regions was more pronounced after lesion of the pelvic plexus. Lesion of the ovarian nerve plexus caused a depletion in the NPY-immunoreactive nerve fibres and a decrease in NPY concentration in the upper part of the uterine horn. Pelvic nerve section, inferior mesenteric ganglia excision and superior ovarian nerve section had no effect on the NPY innervation in the genital tract. Injection of fluorogold into the cervix and lower part of the uterus combined with immunohistochemistry revealed that 87.5% of labelled neurons in the pelvic plexus were NPY-immunoreactive. Following injection of fluorogold into the upper part of the uterus, 92% of labelled neurons in the suprarenal ganglia were NPY-immunoreactive. Treatment with 6-hydroxydopamine revealed that the NPY-immunoreactive nerve fibres were non-noradrenergic in the cervix, but were noradrenergic in the upper part of the uterus. In the uterine body and lower part of the uterine horn, both noradrenergic and non-noradrenergic NPY-immunoreactive nerve fibres were observed. These data demonstrate the major contribution of pelvic plexus neurons in the non-noradrenergic NPY innervation of the lower part of the genital tract, and the involvement of the suprarenal ganglia in the noradrenergic NPY innervation of the upper part of the uterus via the ovarian nerve plexus.     

Distribution of enteric nerve cells projecting to the superior and inferior mesenteric ganglia of the guinea-pig

Retrograde tracing, using Fast Blue dye, was employed to determine the distribution of enteric nerve cells that project to the superior mesenteric and inferior mesenteric ganglia of the guinea-pig. Retrogradely labelled neurons were found in the myenteric but not submucous ganglia. When the superior mesenteric ganglion was injected, labelled neurons were found in low frequencies (less than 5 nerve cell bodies/cm2) in the duodenum, jejunum, ileum, caecum and proximal colon. The distal colon was analysed in five segments of equal length (1-5; oral to anal). Segment 1 had about 4 labelled nerve cells/cm2, whereas segments 2 to 5 displayed an average of about 25 nerve cells/cm2. The rectum contained about 36 labelled neurons/cm2. After injection of the inferior mesenteric ganglia with Fast Blue, no labelled neurons were found in the duodenum, jejunum, ileum or caecum. No labelled cells were observed in the gallbladder. A small number of labelled cells occurred in the proximal colon and in segment 1 of the distal colon. The frequency of labelled cells increased markedly in the more anal regions of the distal colon, and reached a peak in the rectum (138 cells/cm2). Both nerve lesions and immersion of the cut nerve in Fast Blue solution showed that the superior mesenteric nerve carries the axons of neurons located in the middle distal colon to the superior mesenteric ganglion. Almost half of the neurons in the rectum that project to the inferior mesenteric ganglia do so via the hypogastric nerves.(ABSTRACT TRUNCATED AT 250 WORDS)     

Social inhibition of territorial behaviour in yearling male collared lizards, Crotaphytus collaris

The myenteric plexus was investigated in the gastrointestinal tract of pre-diabetic and diabetic non-obese diabetic (NOD) mice. The plexus was immunostained by the avidin-biotin complex method, using a general marker for nerve elements, namely protein gene-product 9.5. The nerve fibres were quantified by point-counting and the number of ganglia and their area were determined by image analysis. The relative volume density of the nerve fibres in duodenal muscularis propria was found to be significantly reduced in of both pre-diabetic and diabetic NOD mice. There was no statistical difference between controls and NOD mice regarding relative volume density of nerve fibres in antral and colonic muscularis propria. The number of myenteric ganglia/mm baseline was significantly decreased in the duodenum of diabetic NOD mice, and showed a non-statistically significant tendency to decrease in pre-diabetic mice. In the antrum and colon, there was no difference between the controls and NOD mice regarding the number of ganglia/mm baseline. Nor was there any significant difference between controls and NOD mice in the area of myenteric ganglia in either antrum, duodenum or colon. It is concluded that the changes in the duodenal myenteric plexus of NOD mice are prior to the onset of diabetes. It is suggested that the absence of changes in the antral and colonic myenteric plexus when using a general marker for neuroelements does not preclude a possible change in cholinergic, adrenergic or peptidergic innervation.     

Absence of the p75 neurotrophin receptor alters the pattern of sympathosensory sprouting in the trigeminal ganglia of mice overexpressing nerve growth factor

Sympathetic axons invade the trigeminal ganglia of mice overexpressing nerve growth factor (NGF) (NGF/p75(+/+) mice) and surround sensory neurons having intense NGF immunolabeling; the growth of these axons appears to be directional and specific (). In this investigation, we provide new insight into the neurochemical features and receptor requirements of this sympathosensory sprouting. Using double-antigen immunohistochemistry, we demonstrate that virtually all (98%) trigeminal neurons that exhibit a sympathetic plexus are trk tyrosine kinase receptor (trkA)-positive. In addition, the majority (86%) of those neurons enveloped by sympathetic fibers is also calcitonin gene-related peptide (CGRP)-positive; a smaller number of plexuses (14%) surrounded other somata lacking this neuropeptide. Our results show that sympathosensory interactions form primarily between noradrenergic sympathetic efferents and the trkA/CGRP-expressing sensory somata. To assess the contribution of the p75 neurotrophin receptor (p75(NTR)) in sympathosensory sprouting, a hybrid strain of mice was used that overexpresses NGF but lacks p75(NTR) expression (NGF/p75(-/-) mice). The trigeminal ganglia of NGF/p75(-/-) mice, like those of NGF/p75(+/+) mice, have increased levels of NGF protein and display a concomitant ingrowth of sympathetic axons. In contrast to the precise pattern of sprouting seen in the ganglia of NGF/p75(+/+) mice, sympathetic axons course randomly throughout the ganglionic neuropil of NGF/p75(-/-) mice, forming few perineuronal plexuses. Our results indicate that p75(NTR) is not required to initiate or sustain the growth of sympathetic axons into the NGF-rich trigeminal ganglia but rather plays a role in regulating the directional patterns of axon growth.     

An ethical responsibility for pain management

Recently, neuronal nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase has been elucidated to be the nitric oxide synthase (NOS) per se. In order to examine the existence and distribution of cerebrovascular nerve fibers containing these substances, NADPH-diaphorase histochemistry was applied to the cerebral blood vessels and the cranial ganglia known to innervate the cerebral vessels in the rat. Numerous nerve fibers with varicosities forming plexuses were observed in the circle of Willis and its branches. In addition, thick nerve bundles were seen to run along the wall of the internal ethmoidal artery. NADPH-diaphorase reaction was prominent in neurons of the sphenopalatine, otic and internal carotid ganglia. This study demonstrated, for the first time, the NADPH-diaphorase-containing nerve fibers in the cerebral vessels and ganglion cells in the parasympathetic and sensory ganglia known to innervate the cerebral vessels.     

Morphometric determination of the methodological criteria for the diagnosis of intestinal neuronal dysplasia (IND B)

Intestinal neuronal dysplasia of the submucous plexus (IND B) is an indicator of a developmental abnormality of vegetative gut innervation. It is the mildest form of an inborn error of intestinal innervation. The diagnosis of IND B does not result in a functional conclusion or clinical recommendation but is often accompanied by oligoneuronal hypoganglionosis of the myenteric plexus or an aganglionosis of the rectum. The aim of this study was to demonstrate by morphometric means a way in which the diagnosis of IND B could be made much more reliable. In 20 control subjects, 40 IND B cases and 10 hypoganglionoses with IND B, it was shown that a specific nerve cell staining (e.g. Lactic dehydrogenase, Succinic dehydrogenase, Diaphorase reaction or an immunohistochemical nerve cell staining) was necessary for diagnosis. Cross sections of giant ganglions and cross sections with large nerve cell numbers (> 7 nerve cell profiles) were the most reliable diagnostic criteria. The morphometric examinations were performed with an optic electronic image analysis system. Biopsy serial sections of the rectum-mucosa that contained submucosa demonstrated that 30-40% of the sections contained no submucous ganglion. Sixty to 70% of the sections showed ganglia of the submucous plexus. In 100 biopsy sections in subjects with IND B, 20 +/- 5% contained giant ganglions cross sections. In the patients with hypoganglionosis of the submucous plexus, 55 +/- 4% sections had no ganglion and 18 +/- 3% had giant ganglion cross sections. The data demonstrate that for a reliable diagnosis of IND B, at least 30 sections are necessary, stained with a dehydrogenase reaction that contain a minimum of 4 giant ganglion cross sections. These data demonstrate that IND B is not a qualitative diagnosis as Hirschsprung's disease but rather a quantitative diagnosis.     

Insulin-stimulated cell growth in insulin receptor substrate-1-deficient ZR-75-1 cells is mediated by a phosphatidylinositol-3-kinase-independent pathway

The projections of enteric neurons to the circular muscle of the guinea pig gastric corpus were investigated systematically by using the retrogradely transported fluorescent carbocyanine dye 1,1'-didodecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI), applied to the muscle layer or myenteric plexus in vitro. DiI-labeled motor neuron cell bodies were located up to 6.3 mm aboral, 17 mm oral, and up to 20 mm circumferential to the DiI application site. Labeled nerve fibers ran for long distances from the DiI application site toward the greater and lesser curvatures, where they coursed parallel to the bundles of the "gastric sling" muscle. The majority of labeled cells were located toward the lesser curvature of the stomach. Nerve cell bodies that were aboral to the DiI application site were usually small, immunoreactive for choline acetyltransferase, and, thus, were likely to be excitatory motor neurons. Neurons that were located orally were larger, fewer in number, and immunoreactive for nitric oxide synthase and, thus, were likely to be inhibitory motor neurons. Application of DiI directly to the myenteric plexus filled neurons up to 15 mm aborally and up to 21 mm orally but labeled few neurons circumferentially. All nerve cells that were filled from either the circular muscle or the myenteric plexus had Dogiel type I morphological features. These results demonstrate a clear polarity of projection of inhibitory and excitatory motor neurons and a functionally continuous innervation of the circular and gastric sling muscle layers. Nonmotor neurons in the myenteric plexus were demonstrated, but neurons with Dogiel type II morphological features are apparently absent.     

Long-term nonprogression in HIV infection. Clinical Epidemiology Group from the Centre d''Information et de Soins de l''Immunodéficience Humaine

Distributions of the cardiac plexuses and cardiac ganglia were gross-anatomically and histologically studied in eight Beijing ducks. The cardiac plexuses consisted of two components, the cardiac nerve arising from the sympathetic trunk and the cranial and caudal cardiac nerves arising from the vagus. Branches of these nerves made the cardiac plexuses on the epicardium. The cardiac plexuses could be divided into the six plexuses, that is, the right and left coronary plexuses, pericardiac transverse sinus plexus, caudal cardiac plexus, and right and left superior cardiac plexuses. There were small ganglia in the caudal cardiac plexus and the right and left coronary plexuses. These ganglia containing multipolar neurons were found like a linking chain in a single nerve.     

Microperoxisome distribution in the central nervous system of the rat

The distribution of microperoxisomes was studied in areas of the central nervous system having high concentrations of catecholaminergic neurons and in areas lacking this neuron type, using the alkaline DAB cytochemical method for catalase. Substantial numbers of microperoxisomes are found in neurons in the locus coeruleus and in nucleus A1 of the medulla, as well as in the substantia nigra, whereas few catalase-reactive bodies are seen in neurons of the cerebrum and cerebellum. The number of catalase-reactive microperoxisomes per unit area in the catecholaminergic neurons of the CNS is comparable to the number seen previously in neurons of the peripheral cervical sympathetic ganglia. Some spinal cord neurons also contain reactive microperoxisomes. Catalase-reactive microperoxisomes are numerous in oligodendrocytes of all areas studied, and in ependymal cells bordering the third and fourth ventricles. Astrocytes contain few reactive structures in the cytoplasm near the nucleus, but they are readily found in astrocytic processes and end-feet.     

Ultrastructure of sea urchin tube feet. Evidence for connective tissue involvement in motor control

An analysis of the ultrastructure of the tube feet of three species of sea urchins (Strongylocentrotus franciscanus, Arbacia lixula and Echinus esculentus) revealed that the smooth muscle, although known to be cholinoceptive, receives no motor innervation. The muscle fibers are attached to a double layer of circular and longitudinal connective tissue which surrounds the muscle layer and contains numerous bundles of collagen fibers. On its outside, the connective tissue cylinder is invested by a basal lamina of the outer epithelium to which numerous nerve terminals are attached. These are part of a nerve plexus which surrounds the connective tissue cylinder. The plexus itself is an extension of a longitudinal nerve that extends the whole length of the tube foot. It is composed of axons, but nerve cell bodies and synapses are conspicuously lacking, suggesting that the axons and terminals derive from cells of the radial nerve. Processes of the epithelial cells penetrate the nerve plexus and attach to the basal lamina. There is no evidence that the epithelial cells function as sensory cells. On the basis of supporting evidence it is suggested that the transmitter released by the nerve terminals diffuses to the muscle cells over a distance of several microns and in doing so affects the mechanical properties of the connective tissue.     

Anatomy of medullary and peripheral autonomic neurons innervating the neonatal porcine heart

Gross and microscopic anatomical investigations were carried out in 14 piglets aged from 4 to 66 days. True Blue (7-50 microliters) and Diamidino Yellow (7-50 microliters) were injected individually into 2 different cardiac sites (the right atrial ganglionated plexus, the inferior vena cava, inferior atrial ganglionated plexus, the right atrium or the right ventricle). Gross anatomy: Globular superior cervical and nodose ganglia, elongated stellate ganglia, multiple small middle cervical ganglia and multiple small mediastinal ganglia along the course of cardiopulmonary nerves were identified. Microscopic anatomy: Neurons innervating specific cardiac regions or intrinsic cardiac ganglionated plexuses were distributed relatively evenly among stellate (primarily in their cranial poles) and middle cervical ganglia bilaterally, fewer labeled neurons being located in the superior cervical and mediastinal ganglia bilaterally. Parasympathetic efferent preganglionic neurons associated with either intrinsic cardiac ganglionated plexus studied were identified primarily throughout the ventrolateral region (the external formation) of the nucleus ambiguus bilaterally. Labeled neurons were also identified throughout the right and left nodose ganglia. Individual neurons did not project axons to different cardiac regions, as no double-labeled neurons were identified. No correlation between age and the numbers and locations of labeled neurons was apparent. Thus, porcine sympathetic efferent neurons which innervate individual cardiac regions, including intrinsic cardiac ganglionated plexuses, lie scattered primarily throughout the right and left mediastinal and middle cervical ganglia as well as the cranial poles of stellate ganglia at birth, apparently changing little during the first 2 months of age. Porcine cardiac parasympathetic efferent preganglionic neurons are located primarily in the external formation of the nucleus ambiguus bilaterally at birth. The numbers of afferent cardiac neurons distributed throughout the nodose ganglia bilaterally also change little during that time. It is concluded that most of the autonomic neurons which innervate the heart are in place at birth.