Substance P-like immunoreactive nerve fibers in the pars distalis of the anterior pituitary in the dog

Summary The pars distalis of the anterior pituitary is known to be regulated by hypothalamic hormones. Recently, we have discovered the presence of substance P-like immunoreactive nerve fibers in the pars distalis of the monkeys. Substance P-like immunoreactivity in the pars distalis of the dog was investigated in this study. A substantial amount of substance P-like immunoreactive nerve fibers with a large amount of varicosities were found. They were widely distributed in the gland, more abundant along its periphery. Most of them were closely related to the glandular tissue, some were located on vascular walls. Substance P-like immunoreactive nerve fibers were also found in the meningeal sheath of the anterior pituitary. They could be followed into the parenchyma of the gland.     

Über Aufbau und Innervation der Kopfanhänge der prosobranchen Schnecken

Zusammenfassung Das Epithel der Kopfanhänge von elf marinen und Süßwasserprosobranchiern besteht aus prismatischen bis kubischen Stützzellen mit meist dichtem Mikrovillussaum und z.T. Pigmentgranula sowie Sinneszellen, die fast immer in Form sekundärer Sinneszellen vorliegen; nur bei Patella coerulea kommen vermutlich auch primäre Sinneszellen vor. Ihr Zytoplasma ist apikal durch glattwandige E. R.-Zisternen, helle Bläschen und Mikrotubuli gekennzeichnet. Außerdem tragen diese Zellen Zilien und stehen basal mit Nervenendigungen in Kontakt, die sich in drei Gruppen einteilen lassen: 1. Vermutlich cholinerge Endigungen mit optisch leeren Bläschen (Ø 600–800 Å). 2. Endigungen mit dense core vesicles (Ø 1000–1100 Å). Die Annahme, daß diese Endigungen biogene Amine enthalten, wird durch fluoreszenzmikroskopische Befunde gestützt. 3. Endigungen mit großen (Ø 3000–4000 Å) neurosekretorischen Elementargranula.

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Structure and innervation of the cephalic tentacles of Prosobranch molluscs

Summary The epithelium of the cephalic tentacles of eleven marine and freshwater prosobranch snails consists of villus bearing supporting cells, which partly contain pigment granules, and sensory cells, which occur in form of secondary sensory cells with the exception of Patella coerulea which presumably possesses primary sensory cells. These receptor cells are characterized as chemoreceptors by apical cilia, smooth surfaced E.R., microtubulues and empty vesicles. At their bases they are in close contact with nerve endings which can be classified in three groups: 1. presumably cholinergic endings with clear vesicles (Ø 600–800 Å). 2. endings with dense core vesicles (Ø 1000–1100 Å). The assumption that these endings contain biogenic amines is supported by positive fluorescence microscopical tests. 3. Endings with big (Ø 3000–4000 Å) neurosecretory elementary granules.

Herrn Prof. Dr. W. Bargmann danke ich für die Überlassung eines Arbeitsplatzes im Anatomischen Institut Kiel.     

Synapse-like contacts between axons of the pineal tract and the subcommissural organ in Rana perezi (Anra) and their absence in Carassius auratus (Teleostei): ultrastructural tracer studies

The present study was designed to investigate the controversial subject of the existence of a neural input from the pineal organ via the pineal tract to the subcommissural organ (SCO) in teleosts and anurans. Horseradish peroxidase was injected into the pineal organ and pineal tract of Carassius auratus and Rana perezi. Within the pinealofugal fibers the tracer was visualized at the light-and electron-microscopic levels either by immunocytochemistry using an anti-peroxidase serum, or by revealing the enzymatic activity of peroxidase. In both species, labeled myelinated and unmyelinated fibers of the pineal tract were readily traced by means of electron microscopy. In R. perezi, numerous terminals contacting the SCO cells in a synapse-like (synaptoid, hemisynaptic) manner bore the label, whereas a different population of endings was devoid of the tracer, indicating that in this species the SCO receives a dual neural input, one of pineal origin, the other of unknown source and nature. In the SCO of C. auratus, neither labeled nor unlabeled synapse-like contacts were found. Thus, in this latter species, a direct neural input to the SCO is missing. It is concluded that the secretory activity of the SCO can be controlled by different mechanisms in different species, and that more than one neural input mechanism may operate in the same species.     

Development of chicken intrafusal muscle fibers

The first sign of developing intrafusal fibers in chicken leg muscles appeared on embryonic day (E) 13 when sensory axons contacted undifferentiated myotubes. In sections incubated with monoclonal antibodies against myosin heavy chains (MHC) diverse immunostaining was observed within the developing intrafusal fiber bundle. Large primary intrafusal myotubes immunostained moderately to strongly for embryonic and neonatal MHC, but they were unreactive or reacted only weakly with antibodies against slow MHC. Smaller, secondary intrafusal myotubes reacted only weakly to moderately for embryonic and neonatal MHC, but 1–2 days after their formation they reacted strongly for slow and slow-tonic MHC. In contrast to mammals, slow-tonic MHC was also observed in extrafusal fibers. Intrafusal fibers derived from primary myotubes acquired fast MHC and retained at least a moderate level of embryonic MHC. On the other hand, intrafusal fibers developing from secondary myotubes lost the embryonic and neonatal isoforms prior to hatching and became slow. Based on relative amounts of embryonic, neonatal and slow MHC future fast and slow intrafusal fibers could be first identified at E14. At the polar regions of intrafusal fibers positions of nerve endings and acetylcholinesterase activity were seen to match as early as E16. Approximately equal numbers of slow and fast intrafusal fibers formed prenatally; however, in postnatal muscle spindles fast fibers were usually in the majority, suggesting that some fibers transformed from slow to fast.     

The innervation of the trachea and extrapulmonary bronchi of the mouse

Summary In the mouse, nerves were located throughout the trachea and extrapulmonary bronchi in both the smooth muscle and the connective tissue. However, no nerves were found within the epithelium. In the smooth muscle there were large numbers of nonmyelinated nerves. These were usually en passant elements but varicosities containing small mitochondria and vesicles were also seen; these axons sometimes appeared to be efferent to the muscle.Unilateral cervical vagotomy reduced the numbers of nerves in the muscle of the trachea and ipsilateral primary bronchus, suggesting that they were afferent. The intramuscular nerves were characterized in terms of their complement of cytoplasmic organelles; in particular nerves containing many mitochondria disappeared following vagotomy.Pretreatment of mice with 5-hydroxydopamine to accentuate the electron-opacity of catecholamine-containing granules resulted in 3.5% of the nerves within tracheal muscle showing such granules.The afferent nerves of the smooth muscle may be complex branching structures with many varicosities. The absence of epithelial nerves may be related to the absence of the cough reflex in the mouse.     

Fine structural and cytochemical study of the innervation of smooth muscle in an amphibian (Bufo marinus) lung before and after denervation

Summary The innervation of the toad (Bufo marinus) lung was studied with transmission electron microscopy and fluorescence techniques, both before and after 12 or 20 days close vagosympathetic denervation. Four cytologically distinct types of neuronal processes were recognised, in relation to the visceral muscles of the lung. These were described as cholinergic, adrenergic, nonadrenergic/non-cholinergic (NANC) and sensory on the basis of the characteristics of their vesicular content and cytochemical reactions. An apparent efferent innervation of visceral smooth muscle was achieved by NANC (50%), cholinergic (25%) and adrenergic (25%) fibres. A few sensory fibres were also present. After denervation only NANC fibres persisted, showing that the cell bodies of these fibres were intrapulmonary. The vascular smooth muscle was supplied by cholinergic, adrenergic and sensory fibres. In the walls of the proximal branches of the pulmonary artery were fibres containing large dense-cored vesicles. These profiles, which were associated with the vasa vasorum, were similar to neurosecretory fibres. After denervation all neural profiles associated with the vasculature had degenerated. The observations suggest that vagal vasodepressor effects in the toad lung are mediated indirectly through relaxation of visceral muscle strands which in their contracted state compress vascular channels.The authors would like to thank Dr. J.R. McLean for technical advice on fluorescence microscopy. This work was supported by a grant from the Australian Research Grants Committee     

Development of innervation to the atrial myocardium of the rabbit

Summary The development of innervation to the atrial myocardium of rabbits from 20th day of gestation to 35 days postnatal was studied ultrastructurally by electron microscopy and by demonstration of catecholamines by histofluorescence. Special attention was directed to the first morphologic appearance of nerve fibers and terminals and the closeness of juxtaposition of terminals with myocardial cells. Adrenergic and cholinergic terminals were identified on the basis of their differential ability to take-up and store the false adrenergic neurotransmitter 5-hydroxydopamine. Adrenergic terminals were first encountered at 20 days of gestation whereas cholinergic terminals could not be positively identified until the 24th day of gestation. Throughout development adrenergic terminals were more numerous than cholinergic, about 71 % of the terminals encountered being adrenergic. Many terminals approach closely (20–30 nm) to the sarcolemma of the muscle cells of the atrium. In many instances adrenergic and cholinergic fibers travel together in the same nerve bundle and are closely apposed without intervening Schwann-cell cytoplasm. Such a relationship could allow peripheral interaction between these fibers in the myocardium.Supported in part by the Kentucky Heart Association, Human Development Studies Program of the University of Kentucky and DHEW Grant 1 RO1 HL 22226-01 HED from the National Heart, Lung and Blood Institute. The technical assistance of Merle Wekstein is appreciated     

Innervation of the pineal gland in the Mongolian gerbil (Meriones unguiculatus)

Summary Parasympathetic stimulation of parotid glands has been studied in vivo, a) in normal resting glands, b) 72 h after post-ganglionic sympathectomy and c) after adrenergic degranulation of the acinar cells.Morphological results in each gland were compared with a similarly pretreated, but not parasympathetically stimulated, contralateral gland from the same animal.On parasympathetic stimulation of glands with densely granulated acinar cells (groups a- and b-) a variable, but usually relatively small, tendency for vacuole formation occurred in some cells. After prior degranulation of the cells (group c-) the tendency for vacuole formation was greatly accentuated. This indicates that the pre-existing metabolic state of the cells can influence the responses to stimulation of a single nerve. Dilatation of rough endoplasmic reticulum and nuclear changes were also more prominent after parasympathetic stimulation of previously degranulated acinar cells, and this suggests that parasympathetic impulses may have strong activating effects on resynthesis under these conditions. It is also likely that parasympathetic stimulation induced some, albeit small, degree of degranulation and, since this occurred in the absence of sympathetic nerves (group b-) it was probably the consequence of a direct cholinergic effect. The present results therefore indicate that the concept of an absolute dichotomy between parasympathetic and sympathetic responses is not tenable in this tissue.Analyses of saliva for amylase and peroxidase gave complex results but indicate that the two enzymes are not necessarily secreted in parallel. The morphological results support the idea that some enzyme molecules may have entered the saliva without being prepackaged into secretory granules, but could have passed directly from dilated cisterns of rough endoplasmic reticulum into intra-cellular vacuoles, and this tendency was most apparent after para-sympathetic stimulation of previously degranulated cells.Travel grants from the Wellcome Trust to J.R. Garrett are gratefully acknowledged. This work has been helped by the technical assistance of Mr. P.S.A. RowleyM.R.C. Research Assistant     

Ultrastructure of arterioles in the cat brain

Summary A total of 110 arterioles were examined in the brains of cats; different sites were studied including the cortex, putamen, pons and crus cerebri. No internal elastic laminae were seen in the subendothelial space, although occasional fragments of elastic material were present in the larger arterioles. The media was composed of one, two or three layers of smooth muscle cells which interlocked in such a way that the vessel wall thickness was constant. Numerous tight junctions were seen between adjacent smooth muscle cells and between the endothelium and smooth muscle cells. Apart from the usual cell organelles, the smooth muscle cells of arterioles had numerous dense patches on the cell surface. The structure of the adventitia varied according to the diameter of the vessel and the site in the brain; it contained adventitial cells, bundles of collagen fibres and nerve fibres. Innervation of arterioles was more constant in the brain stem than in the cortex. Metarterioles had less specialised, atypical smooth muscle cells, a discontinuous media and numerous, extensive myoendothelial tight junctions; they were not innervated by nerve fibres. The diameter of metarterioles was less than 10 m whereas that of arterioles was 10–45 m. The possible functional aspects of arteriolar innervation are discussed.     

Ultrastructure of the basiepithelial nerve plexus of the sea urchin,Centrostephanus longispinus

Summary In submandibular glands of rabbits both adrenergic and cholinergic axons are intimately associated with parenchymal cells of the intercalary ducts and the granular tubules, lying beneath the basement membrane and often in the space between the parenchymal cell and an associated myoepithelial cell. The submandibular acini receive a less intimate and less plentiful innervation by adrenergic and cholinergic axons which remain outside the basement membrane and are still associated with Schwann cells. Occasional axons of both adrenergic and cholinergic type occur beneath the basement membrane of submandibular striated ducts in intimate association with basal parts of the cells.In the parotid glands numerous adrenergic and cholinergic axons are found beneath the basement membrane of acini and intercalary ducts in intimate association with the cells.This work has been helped by the technical assistance of Mr. P.S.A. Rowley