Histochemical study on the nerve endings in the lung of Rattus rattus rufescens and Francolinus pondicerianus

Presence and the relation of the nerve endings with associated structures in the lund of Rattus rattus rufescens (Indian black rat) and Francolinus pondicerianus (grey partridge or safed teeter) has been studied by cholinesterase technique. Dot, plate, and Vater pacini Corpuscles like nerve endings in the lung of Rattus and dot, and bulb like nerve endings with axis cylinder covered with myelinated sheath in the lung of Francolinus were recorded. The nerve endings were AChE-positive.     

Fine structure of nervous elements in the pancreas of some vertebrates

Summary The innervations of the exocrine and endocrine pancreas of some vertebrate animals were studied by electron microscopy. The pancreas of the bat and monkey contained ganglion cells in the interlobular connective tissue or between acinar cells. Unmyelinated nerve fibers ran through the interlobular connective tissue and reached the exocrine and endocrine parts, and terminated there as the endings. The nerve endings within the pancreas could be divided into four types: 1. Type 1-a of the nerve ending contained only agranular synaptic vesicles of about 500 Å in diameter. 2. Type 1-b characterized by containing agranular synaptic vesicles and some large cored vesicles (1,000 Å diameter). These two types of nerve endings might belong to the cholinergic (parasympathetic) endings. 3. Type 2-a contained small cored vesicles and agranular synaptic vesicles along with a few large cored vesicles. 4. Type 2-b was characterized by containing vesicles of the same size as those of agranular synaptic vesicles, and a majority of these vesicles contained bar-shaped crystalloids. This ending also contained a few large cored vesicles. These nerve endings of Type 2-a and 2-b might be the adrenergic (sympathetic) endings.     

Characteristics of electrical activity in different sections of nerve endings in the frog

In experiments on frog sartorius neuromuscular preparations, the evoked electrical responses of nerve endings were recorded by extracellular microelectrodes. It was shown that in proximal parts of the nerve ending, the three-phase response (+ - +) with a high amplitude negative phase occurred due to motor nerve stimulation. With movement of the extracellular electrode in distal direction a certain increase of the initial positive phase and a significant decrease of the negative one were observed. At the end of the terminal that response transformed to the monophasic one (+). On local iontophoretic application of tetrodotoxin (TTX) to the recording site two components of the nerve ending response were revealed: TTX-insensitive and TTX-sensitive. A significant decrease of the TTX-sensitive component occurred along the course of the nerve ending. That component was absent from the distal synaptic areas. It is concluded that in frog nerve ending, the action potential propogates with decrement while depolarization of the end parts of the terminal is passive in nature.     

Fine Structure of Subepithelial “Free” and Corpuscular Trigeminal Nerve Endings in Anterior Hard Palate of the Rat

Axonally transported protein labeled many trigeminal nerve endings in subepithelial regions of the anterior hard palate of the rat. Sensory endings were most numerous in the lamina propria near the tips of the palatal rugae where large connective tissue and epithelial papillae interdigitated. Two kinds of sensory ending were found there: “free” endings, and a variety of corpuscular endings. The “free” sensory endings consisted of bundles of unmyelinated axons separated from the connective tissue by relatively unspecialized Schwann cells covering part or all of their surface and a completely continuous basal lamina; they were commonly found running parallel to the epithelium or near corpuscular endings. The corpuscular sensory endings all had a specialized nerve form, specialized Schwann cells, and axonal fingers projecting into the corpuscular basal lamina or connective tissue. There were at least four distinct types of corpuscular ending: Ruffini-like endings were found among dense collagen bundles, and they had a flattened nerve ending with a flattened Schwann lamella on either side. Meissner endings had an ordered stack of flattened nerve terminals with flattened Schwann cells and much basal lamina within and around the corpuscle. Simple corpuscles were single nerve endings surrounded by several layers of concentric lamellar Schwann processes. Glomerular endings were found in lamina propria papillae or encircling epithelial papillae; they were a tangle of varied neural forms each of which had apposed flattened Schwann cells, and a layer of basal lamina of varied thickness. Fibroblasts often formed incomplete partitions around Meissner and simple corpuscles.

The axoplasm of all kinds of subepithelial sensory endings contained numerous mitochondria and vesicles, as well as occasional multivesicular bodies and lysosomes; the axoplasm of all endings was pale with few microtubules and neurofilaments. The specialized lamellar Schwann cells had much pinocytotic activity. Four kinds of junctions were found between the corpuscular sensory endings and the lamellar Schwann cells: (1) symmetric densities that resemble desmosomes; (2) asymmetric densities with either the neuronal or glial membrane more dense; (3) neural membrane densities adjacent to Schwann parallel inner and outer membrane densities; and (4) sites of apparent Schwann endocytosis associated with neural blebs. The “free” sensory endings only made occasional desmosome-like junctions with their Schwann cells.

These observations are discussed in relation to possible mechanosensory transduction mechanisms, with particular attention to axoplasmic structure, axonal fingers, and neural and nonneural cell associations.     

Ultrastructure of Merkel corpuscles and so-called "transitional" cells in the white Leghorn chicken

In the chicken Merkel corpuscles are located in the dermis and consist of specialized Merkel cells, discoid nerve endings and lamellar cells. Merkel cells contain characteristic membrane-bound dense-core granules and bundles of microfilaments. Asymmetric junctions, synapse like, with thickened membranes and clusters of dense-core vesicles were observed between the Merkel cells and the nerve endings. The nerve ending is derived from myelinated nerves and sometimes contains clusters of clear vesicles. A laminar system formed by lamellar cells of the Schwann cell type encloses the Merkel cells and the nerve endings. So called "transitional" cells, showing some of the morphological features of both keratinocytes and Merkel cells, were observed in the basal layer of the epidermis. One was located partly in the epidermis and partly in the dermis. The structure of Merkel corpuscles is compared with that of Merkel cells in other tetrapods. The developmental significance of "transitional" cells and the origin of Merkel cells are discussed.     

Incorporation of radioactive glucosamine into macromolecules at nerve endings

Mice were injected intracerebrally with [¹⁴C]glucosamine, and incorporation into macromolecules in various subcellular fractions of brain was studied at a number of times after administration of the precursor. The [¹⁴C]glucosamine was rapidly incorporated into macromolecules of all the subcellular fractions of brain including both the soluble and particulate fractions of isolated nerve endings. Incorporation into macromolecules in the soluble fraction of nerve endings was quite extensive 3 hr after administration of the precursor and the specific acitvity of this fraction fell thereafter. In contrast there was only slight incorporation of [¹⁴C] leucine into the soluble protein from isolated nerve endings in the first few hours after administration, whereas the other subcellular fractions were maximally labelled at that time. The data suggests that, unlike protein which is largely transported to nerve endings in the axoplasm, there is extensive incorporation of carbohydrate into macromolecules in nerve endings. Whereas the protein component of a glycoprotein or mucopolysaccharide may be transported to the nerve ending from the perikaryon, the structure and function of this protein may be modified at the nerve ending by further incorporation of glucosamine, sialic acid and possibly other carbohydrates. The carbohydrate-containing macromolecules could influence nerve ending function immediately after these final synthetic reactions since these reactions occur at the nerve ending and not in the perikaryon.     

A histochemical study of the neuro-muscular spindles in the tongue of some rodents.

An investigation was undertaken to study the Neuro-muscular spindles in the tongue (Rattus rattus rufescens and Suncus murinus) by cholinesterase technique, under maintained pH of 5.2, incubation period 20 h., and temperature 38 degrees C (Rattus) and 39 degrees C (Suncus). Neuro-muscular spindles were observed frequently distributed from anterior to posterior of the tongue and took a little darker stain than the ordinary muscle fibres (negatively stained) while the nerve fibres and ganglia took still darker stain. Nerve fibres (myelinated, non-myelinated and tortuous) were seen penetrating and embracing the muscle spindles and most of them ran parallel to the spindles. Ganglia of various shapes (oval, irregular, elongated and rounded) and sizes (large, Rattus and medium and small, Suncus) were AChE-positive. Ganglia small and medium in size, rounded and elongated in shape were arranged in chain-like fashion on the muscle spindles in Suncus (Fig. 9, 10). The blood vessels (artery, vein, and capillary) were recorded either in close association with the muscle spindles or lying parallel to it. In all cases they were closely related and deeply associate either with the nerve fibres of the plexus of the muscle spindles or with the nerve fibres of the neural network of vessel. Perivascular and perimuscular ganglia with pre- and postganglionic nerve fibres were closely related to each other. Nerve endings, dot-like (Rattus) and bulb-like (Suncus) were occasionally recorded. AChE activity was found randomly in the muscle spindles (Fig. 2, 5.) and in the form of dark brown patches in the ordinary muscle fibres.     

Fine structure of islet-cell innervation in the pancreas of normal and alloxan-treated rats

Summary The innervation of the islets of Langerhans of normal albino rats and of albino rats treated with several daily doses of 125 mg/kg of alloxan was studied by electron microscopy. In the normal rat, nerve endings containing either agranular vesicles (200–400 Å) alone or in combination with large granular vesicles (500–800 Å) were found on both alpha and beta cells. Infrequently a third type of nerve ending containing small granular synaptic vesicles could be observed. Bundles of unmyelinated axons were also seen, as were typical autonomic ganglion cells. Similar normal neural elements were noted in rats treated with alloxan. However, islets of alloxan-treated animals also possess large elliptical profiles which appear to be dystrophic nerve terminals. These structures most frequently contact degranulated beta cells. Islets of Langerhans fixed with zinc iodide-osmium (ZIO) reported to specifically impregnate synaptic vesicles were also studied. Synaptic vesicles of normal axons and nerve endings as well as of the dystrophic structures were filled with ZIO reactive material. These studies suggest that alloxan may induce autonomic nerve ending changes in the rat endocrine pancreas. This may result from neuronal hyperactivity in an attempt to secrete insulin from the post-alloxan insulin-depleted beta cell.     

Changes in the ultrastructure of Merkel's nerve endings of the sinus hair of rats after denervation

Merkel nerve endings of sinus hairs in the upper lip of the rat were observed after having cut the nervus infraorbitalis. The nerve terminal of the Merkel nerve ending has already been degenerated 24 hours after denervation and was phagocytised by neighbouring keratinocytes. The Merkel cells did not change in structure after having lost their nerve terminal even within 169 days after nerve crush. Their position in the stratum basale of the sinus hairs remained constant; the number, size and position of the osmiophilic granules in the cytoplasm of the Merkel cells did not change. These results may show, that Merkel cells are not modified keratinocytes. The possibilities of their origin are discussed.     

Neuro-histochemical study of the tongue of Gallus domesticus (White Leghorn)

An investigation was undertaken to study the neuro-histochemistry of tonue of Gallus domesticus (white Leghorn) by cholinesterase technique under maintained pH of 5.2, temperature 40 degrees C and incubation period 20 h, and Ungewitter's silver impregnation method. Fungiform (8--16 in number), filiform (5--16 in number), circumvallate and foliate (numerous) were recorded profusely innervated in the various regions of the tongue. Neuro-muscular spindles related either with the non-myelinated, myelinated nerve fibres or with the ganglia via pre- or postganglionic nerve fibre showed cholinesterase (AChE) positive activity in the form of dark brown patches. Neural network over the glands, formed the basket envelope likestructures. Glands showed positive cholinesterase (AChE) reaction on the periphery. Dot-like nerve endings were connected with the neural terminal network in the foliate papillae region and motor end plate like endings on blood vessel, were occasionally recorded. Parasympathetic ganglia of various shape and sizes, arranged in chain-like fashion were observed in the connective tissue space between the fungiform and circumvallate papillae and foliate papillae region. The pre- and postganglionic fibres of these ganglia were related with the anteriorly and posteriorly located papillae via the neural terminal network. Cholinesterase (AChE) activity was marked either in the centre, of the fungiform or on the periphery of the filiform, circumvallate and foliate papillae.     

Submicroscopic Changes of the Nerve Endings in the Adrenal Medulla after Stimulation of the Splanchnic Nerve

The nerve endings of the adrenal medulla of the rabbit were studied under the electron microscope in the normal condition and after prolonged electrical stimulation of the splanchnic nerve. With a stimulus of 100 pulses per second for 10 minutes, there is an increase in the number of synaptic vesicles in the nerve ending. The mean number is of 82.6 vesicles per square micron in the normal and of 132.7 per square micron in the stimulated glands. With a stimulus of 400 pulses per second for 10 minutes, there is a considerable depletion of synaptic vesicles and other changes occur in the nerve endings. The mean number of vesicles is of 29.2 per square micron. These results are interpreted as indicative of an increased activity of the ending in one case, and as a diminished activity and fatigue of the synaptic junction in the other.     

Further studies of the transport of protein to nerve endings

Mice were injected intracerebrally with [l-¹⁴C]leucine, and the specific activities of subcellular fractions of brain and effractions of isolated nerve endings were determined. There was a progressive increase in the specific activity of protein associated with isolated nerve endings after incorporation of [l-¹⁴C]leucine into whole brain protein had terminated. Although, the incorporation of [¹⁴C]leucine into soluble protein of whole brain did not differ significantly in mice which were 3 months or 1-year old, the subsequent increase in specific activity of soluble protein isolated from nerve endings was significantly greater in the younger animals; 6-month-old mice were intermediate. Therefore, changes in some aspect of the transport of protein to nerve endings is altered even after sexual maturity. Anaesthetization with pentobarbitone during incorporation of [¹⁴C]leucine into protein, and inhibition of protein synthesis with acetoxycycloheximide after incorporation of [¹⁴C]leucine was complete, did not interfere with the subsequent appearance of radioactive protein at the nerve ending. Evidence is presented for the transport, from a proximal site of synthesis, of protein associated with particulate components of the nerve ending, including synaptic vesicles.     

Demonstration and localization of synapsin I in vestibular receptors in the cat: immunocytochemical study

The presence and localization of synapsin I, a neuron-specific phosphoprotein, was investigated in the cat vestibular epithelium, using a rabbit antisynapsin I anti-serum. The staining was performed by immunofluorescence or by a peroxidase-antiperoxidase (PAP) technique. A strong immunoreactivity was observed with both methods. This immunoreactivity appeared as spherical patches distributed in the lower part of the epithelium. This distribution pattern is very similar to that of the efferent synaptic endings which form axodendritic synapses with the afferent nerve chalice of type I hair cells, or axosomatic synapses with type II hair cells. Some of the nerve chalices were also labelled; in this case, the immunoreactivity was more evident with PAP staining. These results thus suggest the presence of large amounts of synapsin I in the vestibular efferent nerve endings. These endings are known to be filled with numerous synaptic vesicles. This localization of synapsin I is well correlated with previous work that report a close association between synapsin I and small synaptic vesicles. The presence of synapsin I in sensory endings such as the afferent nerve chalices was unexpected and is under investigation.     

Effect of armin on the ultrastructure of the neuromuscular synapse]

Alterations induced by the cholinesterase inhibitor armin (5.10(-7) g/ml) in the ultrastructure of motor nerve endings of the rat phrenic diaphragmal preparations at rest or electric stimulation of the nerve were studied. It was shown that armin at rest induced ultrastructural lesions in the endings similar to those in the control preparations during nerve stimulation. Electric stimulation did not produce additional changes in the ultrastructure of the neuromuscular junction under armin action. It is suggested that the disorder of the nerve ending function may be of importance in the mechanism of the blocking action of armin on the neuromuscular transmission.     

Histological study of motor innervation to long nuclear chain intrafusal fibers in the muscle spindle of the cat

Several muscle spindles of the cat tenuissimus muscle were cut in serial, 1-micron thick transverse sections and stained with toluidine blue in search for long nuclear chain intrafusal muscle fibers. Five complete poles of the long chain fibers were located. Each fiber pole displayed one plate-type motor ending situated in the extracapsular fiber region. The endings were supplied by myelinated motor axons that originated from intramuscular nerve fascicles containing motor axons to extrafusal muscle fibers. One of the endings was innervated by a collateral from a motor axon that supplied an extrafusal end-plate. Ultrastructurally, the long chain endings resembled extrafusal end-plates. They were more complex, in terms of prominence of sole-plate and degree of post-junctional folding, than any other intrafusal ending present in the spindles. The motor endings of the long chain fibers were assumed to be the terminals of static (fast) skeletofusimotor axons, which preferentially innervate the longest nuclear chain fibers of cat muscle spindles.     

Cytosolic sialidase from the nerve endings of developing rat forebrain

The developmental profile of cytosolic sialidase from the nerve ending and the nerve ending-free compartments of rat forebrain was studied from birth to 150 days. Soluble extracts containing the cytosol from nerve endings and nerve ending-free tissue were separately prepared; the recovery of cytosol in the two extracts was followed by assaying lactate-dehydrogenase. In both cytosolic extracts the content of sialidase and lactate-dehydrogenase had a marked and progressive enhancement from the 5th to the 20th day of life and then maintained a constant level through adult life. However the rate of lactate-dehydrogenase and sialidase increase in the two cytosolic compartments was different. Lactate-dehydrogenase increased at a very similar rate in the two cytosols. Instead the rate of sialidase increase was greater in the cytosol from nerve endings from the 5th to the 10th day of life, and, inversely, in the cytosol from nerve ending-free tissue from 10 to 20 days. Between the 5th and 10th day of life the nerve ending cytosol underwent an enrichment of sialidase which was several times higher than that of lactate-dehydrogenase; in the other periods of forebrain development sialidase and lactatedehydrogenase moved in parallel.The variations with age of cytosolic sialidase from the nerve ending-free tissue seemed to follow the overall process of brain development. The nerve ending soluble sialidase would more specifically reflect formation of synaptic junctions in some brain regions in a well defined period of brain maturation.     

Submicroscopic morphology of the infrared receptor of pit vipers

Summary The fine structure of the infrared receptor membrane of pit vipers has been studied under the electronmicroscope. From the outer to the inner surfaces, within a total thickness of only 8 to 16 , the following seven layers were recognized: 1. Outer epithelium, 2. outer connective layer, 3. layer of vacuolar cells, 4. layer of nerve endings, 5. layer of nerve fibers, 6. inner connective layer, 7. inner epithelium.The nerve endings, which form a densely packed layer, represent the most prominent component of the sensory membrane. Their inner structure is remarkable because of the high mitochondrial concentration. The population density of these organoids is as great as virtually to occupy the entire ending. Almost half of the volume of the sensory membrane is thus made of compact masses of mitochondria.The structure of the myelinated nerve fibers entering the sensory membrane, was analyzed together with the stages of transformation into nerve endings.This study revealed that there is a special region of the nerve fiber in its transition toward the nerve ending where mitochondriogenesis is very active, permitting the analysis of the mechanism of formation of these cell organoids. Some physiological implications inferred from the particular structure of the sensory membrane are discussed. Special emphasis is put on the enormous mitochondrial concentration at the nerve endings. The hypothesis is advanced that these organoids might in some way be involved in the mechanism of transducing emperature changes into nerve impulses.This paper was supported by a grant of the Office of Scientific Research of the U.S. Air Force given to the Instituto de Anatomía General y Embriologia, Facultad de Ciencias Médicas, Universidad de Buenos Aires, Argentina.Postdoctoral fellow of the Instituto Nacional de Microbiología, Buenos Aires, Argentina.     

Morphometric study of the neuromuscular synapses in the adult rat with special reference to the remodelling concept

There is increasing morphologic evidence that neuromuscular synapses are not rigid structures in the mature muscles of adult animals. On the contrary, they may be submitted to a continuous process of remodelling. In silver-impregnated sternocleidomastoid muscles of the young adult rat, we measured synaptic parameters such as nerve terminal length, the number of branching points of terminal arborization, and muscle fiber diameter, and used a morphometric approach to explore specific questions concerning neuromuscular remodelling. Quantitative data indicate that: (a) The complexity and maturation of the nerve endings in this muscle are very variable and the increase in branching points is not paralleled by an increase in terminal length; (b) Muscle fiber diameter is related only marginally to presynaptic parameters; (c) Accessory ending formation occurs when the original ending does not reach the mean size of endings in singly innervated areas; (d) The complexity of individual endings at dually innervated junctions is smaller than the mean development of singly innervated synapses, indicating the existence of some mutual inhibitory influence between closely spaced endings. Morphometric results suggest a continuous process of synaptic formation in this adult muscle.     

The sensorineural apparatus of the human eyelid

The present study describes a complex array of sensory nerve terminals in the human eyelid. In many respects this pattern of innervation resembles that previously described in the rhesus monkey, but in other respects it is unique to man. The most prominent nerve terminals are a complex array of lanceolate and circular Ruffini and free nerve endings that envelop the eyelashes. In addition, Merkel cells have not been conclusively identified to date in other nonhuman primate nonsinus hairs. The external root sheath collar contains Merkel cells, and dermal Merkel cells have also been identified close to the collar. The anterior cutaneous surface of the eyelid contains small vellus hairs with typical lanceolate, Ruffini, and free nerve endings resembling those of primate facial skin. Scattered Meissner and scant simple corpuscles as well as scattered free nerve endings (FNEs) can be identified on the occlusal surface of the eyelid. Intraepithelial as well as dermal FNEs were most easily identified in this region in areas lacking other corpuscular receptors. Corpuscular receptors are especially common at the occlusal/conjunctival angle. The inner or conjunctival surface of the eyelid appears to be a glandular epithelium, whereas in the rhesus monkey it is stratified squamous epithelium. This epithelium needs additional study. In summary, the present study confirms the unique sensory neural status of the human eyelid and verifies the presence of Ruffini nerve terminals by light and electron microscopy and of free nerve ending terminals at least by light microscopy, as well as a unique pattern of innervation of the human eyelash.     

RAPID TRANSPORT OF FUCOSYL GLYCOPROTEINS TO NERVE ENDINGS IN MOUSE BRAIN

Abstract— Mice were injected intracerebrally with mixtures of [³H]fucose and [¹⁴C]gluco-samine, and incorporation into macromolecules in various subcellular fractions of brain was studied at 1, 2, 3 and 4 h after administration of the precursors. There was a lag of several hours between the incorporation of [³H]fucose into the glycoproteins of the whole brain fractions and of that into the soluble and particulate glycoproteins of the nerve ending fractions. In contrast, no lag was observed between the incorporation of [¹⁴C]glucosamine into the macromolecules of the whole brain fractions and of that into the soluble macro-molecules of the nerve ending fraction. We conclude that fucosyl glycoproteins of the nerve ending fraction were synthesized in the nerve cell bodies and transported to nerve endings by rapid axoplasmic transport, whereas a substantial proportion of the glucosamine in the soluble macromolecules of the nerve ending fraction was incorporated by the nerve endings themselves. In addition, our evidence indicates that cyclobeximide inhibited fucose incorporation into brain glycoproteins by inhibiting the synthesis of acceptor proteins rather than fucosyl transferase.