Small granule-containing cells of the central nervous system of the bivalve mollusk Patinopectin yessoensis (Jay)

In the CNS of the Patinopecten yessoensis (Jay) two types of cells have been revealed. The I type cells are typical unipolar neurons with a developed granular endoplasmic reticulum and Golgi compex, with a nucleus containing small amount of chromatin. They possess elementary peptidergic granules. The II type cells have in their cytoplasm and processes a large amount of electron-opague granules, specific for adrenergic systems. The nucleus is rich in clustered chromatin, the granular endoplasmic reticulum is poorly developed, cytosomes are absent. According to their ultrastructural organization the latter correspond to small granular cells of the mammalian autonomic nervous system.     

Dark and light neurons in the cortical plate of the forebrain of the swamp turtle]

Electron microscopic study of the neuronal composition of Emys orbicularis cortical zones as compared with the light microscopy data has revealed that the distinctions in the cell cytoplasm ultrastructure with respect to the dendrite branching pattern and the peculiarities of their localization in the cortical plate are suggestive of a possible metabolic heterogeneity and can serve as a foundation of the morpho-functional characteristics of the neurons. A few neuronal types of neurons have been revealed: 1. light neurons distinguished by the cytoplasm matrix devoid of any electron density, a poorly developed endoplasmic reticulum and a few cytoplasmatic organells and inclusions; 2. light neurons having slight osmophility of the cytoplasm matrix, a more developed endoplasmic reticulum with elongated cisternae, a great number of organells including neurofilaments; 3. dark neurons having a high degree of cytoplasm matrix and the nucleus, a large number of the nuclei, cytoplasm, dendrites and axons in contrast to the above described patterns.     

Annulate lamellae and whorl bodies in the diffuse supraoptic nucleus of the hamster

The ultrastructure of neurons of the diffuse supraoptic nucleus of the hamster has been studied. These neurons show two specializations of the endoplasmic reticulum: annulate lamellae and whorl bodies. From one to three whorl bodies are found in the same neuron. The annulate lamellae and the whorl body cisterns are continuous with the cisterns of the rough endoplasmic reticulum. These neurons present an extraordinarily developed rough endoplasmic reticulum, small mitochondria, neurosecretory vesicles and a Golgi complex filled with electron-dense material. Astrocytic processes of different thickness surround the neurosecretory cells.     

The neurocytes of the caudal part of the tuberomamillary nucleus in Ovis aries L. (light microscopy and electron microscopy studies)

The neurons of the pars caudalis nuclei tuberomammillaris (pc-NTM) were studed light-microscopically and electron-microscopically in sheep and rams of Merino breed. In our study we observed: In the regarded neural nucleus, there is the majority of the great neurons (up to 60 microns in diameter) rich in the NISSL's bodies. When stained with the cresyl violet, the NISSL's substance is apparently stored mainly in peripheral area of the cell body and in the distant parts of numerous protoplasmic processes, what evokes an impression of the "jagged" surface of these cells. After staining with paraldehyde fuchsin, we found purple coloured lumps of irregular shape stored outside the cell bodies, in the neuropil. The less extended cells, usually with lower content of NISSL bodies, are in pc-NTM less frequent. In the electron-microscopic study we identified 3 types of neurons: Cells rich in rough endoplasmic reticulum; "light" cells, "dark" cells. The cells of the 1st type were the most frequent ones. Cisterns of rough endoplasmic reticulum in the 1st type of cells are often dilated. The protoplasmic processes of these cells are frequently stepped over by flat tubuli of endoplasmic reticulum. The 2nd type of cells is characterized by the light cytoplasmic matrix, low quantity of endoplasmic reticulum and frequent occurrence of lipofuscin bodies. The 3rd type of cells are characterized by the high density of cytoplasmic matrix, well developed GOLGI complex, and very broad cisterns of endoplasmic reticulum, forming a labyrinth, and it is bound to a broad perinuclear space.     

The lateral cervical nucleus in the cat

Summary Profiles of 14 neurons all sectioned through the nucleolar plane and 87 isolated dendritic profiles have been analyzed with respect to the surface area covered by boutons and astroglial processes. This analysis has revealed two different types of neurons within the lateral cervical nucleus (LCN) of the cat. The cell types also differ in other ultrastructural respects. One type, which probably consists of projection neurons, is characterized by a rather large size, a relatively small nucleus, numerous mitochondria, well developed granular and agranular endoplasmic reticulum. The cell membrane of these cells shows somatic spines and the perikaryon is covered with boutons to a mean extent of 42%. The other cell type, which probably is internuncial, is smaller, has a proportionally larger nucleus, few mitochondria and a poorly developed granular and agranular endoplasmic reticulum. These cells show no somatic spines and the perikaryal membrane is covered with boutons to an extent of about 10%. Also the bouton populations contacting the two cell types differ from one another. The proportion of internuncial neurons within the LCN has been estimated to about 8%. The internuncial neurons seem to have no preferential localization.The primary dendrites of the projection neurons have a bouton covering of about 48%. No proportional differences in covering could be revealed between different sizes of dendrites.The results are discussed in relation to what is known about the anatomical and physiological organization of the LCN, and also compared with the results obtained in other similar investigations on other parts of the central nervous system.The author is grateful to fil.kand. Göran Engholm for his help with the statistical considerations.This work was supported by grants from Anders Otto Swärds Stiftelse, Stiftelsen Lars Hiertas minne, Åhlén och Holms stiftelse, Åke Wibergs stiftelse and the Swedish Medical Research Council (Project No B70-12X-2710).     

Location matters: the endoplasmic reticulum and protein trafficking in dendrites

Neurons are highly polarized, but the trafficking mechanisms that operate in these cells and the topological organization of their secretory organelles are still poorly understood. Particularly incipient is our knowledge of the role of the neuronal endoplasmic reticulum. Here we review the current understanding of the endoplasmic reticulum in neurons, its structure, composition, dendritic distribution and dynamics. We also focus on the trafficking of proteins through the dendritic endoplasmic reticulum, emphasizing the relevance of transport, retention, assembly of multi-subunit protein complexes and export. We additionally discuss the roles of the dendritic endoplasmic reticulum in synaptic plasticity.     

Ultrastructural aspects of the paraventricular nuclei in the rat

Summary The ultrastructural aspects of the paraventricular nucleus and its neuropil are described in the normal rat.Two types of neurons can be distinguished morphologically. The first type contains numerous dense-core vesicles (mean diameter: 140 nm). The cisternae of the endoplasmic reticulum are arranged parallely at the periphery of the cell body.The second type of neuron contains a few dense-core vesicles (mean diameter: 75 nm) and the endoplasmic reticulum is randomly distributed in the cytoplasm. In the neuropil, two types of dense-core vesicles are observed in separated axons. The histogram of the distribution of their mean diameter clearly indicates a double population of vesicles.The signification of the second type of neuron in the paraventricular nucleus is discussed and its possible relation to TRF synthesis is evoked.This work was supported by a grant from the Belgian National Fund for Scientific Research.The author wish to thank Mrs. Hunninck-Couck for her devoted and skillful technical assistance.     

An electron microscope study of the effects of acute ischaemia in the brain

The structural changes in the CNS of cats subjected to periods of 10 and 20 min of eschaemia have been studied. After 10 min of ischaemia the neurons show loss of granular endoplasmic reticulum and vacuolation of the persisting endoplasmic reticulum. After 20 min of ischaemia there is neuronal cell death, some synaptic degeneration. Glial foot processes swell and occlude the lumen of vessels. The significance of these changes is discussed.     

Ultrastructural features and acetylcholinesterase histochemistry of the rat habenular complex

The ultrastructural features and the acetylcholinesterase (AChE) localization of the rat habenula have been studied. On the basis of different morphology and AChE content, it is suggested that at least two types of neurons are present in the medial habenula (MHb) and three types of neurons in the lateral habenula (LHb). In particular, actively AChE-synthesizing neurons have been noticed in both LHb and MHb. Some unusual ultrastructural arrangements of the endoplasmic reticulum have been described in habenular neurons. Finally, the most common types of synaptic contacts present in the habenular complex have been surveyed.     

Peripheral membrane proteins of sarcoplasmic and endoplasmic reticulum. Comparison of carboxyl-terminal amino acid sequences

Peripheral endoplasmic reticulum membrane proteins residing in the lumen of the endoplasmic reticulum occupy the same space as other secreted proteins. The presence of a four amino acid salvage or retention signal (KDEL-COOH = Lys-Asp-Glu-Leu-COOH) at the carboxyl-terminal end of peripheral membrane proteins has been shown to represent a signal or an essential part of a signal for their retention within the endoplasmic reticulum membrane. In heart and skeletal muscle, a number of sarcoplasmic reticulum proteins have recently been identified which are peripheral membrane proteins. The high-affinity calcium-binding protein (55 kilodaltons (kDa] appears to conform to the above described mechanisms and contains the KDEL carboxyl-terminal tetrapeptide. Thyroid hormone binding protein is present in the sarcoplasmic reticulum, in addition to its endoplasmic reticulum location, and has a modified but related tetrapeptide sequence (RDEL = Arg-Asp-Glu-Leu), which also probably functions as the retention signal. Calsequestrin and a 53-kDa glycoprotein, two other peripheral membrane proteins residing in the lumen of the sarcoplasmic reticulum, do not contain the KDEL retention signal. The sarcoplasmic reticulum may have developed a unique retention mechanism(s) for these muscle-specific proteins.     

Acetylcholinesterase activity and type C synapses in the hypoglossal, facial and spinal-cord motor nuclei of rats. An electron-microscope study

Using the electron-microscope technique of Lewis and Shute, we studied the localization of the acetylcholinesterase (AChE) activity in the hypoglossal, facial and spinal-cord motor nuclei of rats. The technique used selectively detects synapses with subsynaptic cisterns (type C synapses) as well as heavy deposits of reaction products in the rough endoplasmic reticulum, in fragments of the nuclear envelope, in some Golgi zones and on parts of the pericaryal plasma membrane, the axolemma and the dendritic membrane. In C synapses, AChE activity was located in the synaptic cleft and on the membrane of presynaptic boutons. Some C synapses exhibited distinct synaptic specialization in the form of multiple 'active zones'. These zones were characterized by dense presynaptic projections, short dilations of the synaptic cleft, and postsynaptic densities localized between the postsynaptic membrane and the outer membrane of the subsynaptic cistern. Within the postsynaptic densities, rows of rod- or channel-like structures were observed. The subsynaptic cisterns were continuous with the positive rough endoplasmic reticulum. The results are discussed in terms of the possible role of C synapses in the regulation of AChE synthesis in postsynaptic cholinergic neurons and/or in the regulation of AChE release into the extracellular space as well as in the establishment of new synaptic contacts.     

Ultrastructure of the “smooth endoplasmic reticulum cell” in the nucleus recessus lateralis of Salmo gairdneri Richardson

Summary The fine structure of one of the cellular components of the nucleus recessus lateralis of trout is described. These cells have an enormously developed smooth endoplasmic reticulum which makes their identification easy. The relationship of this cell type with the ventricular cavity, neighbouring cells and the presence of synaptic contacts with these cells are also described.     

Some observations on the fine structure of light and dark cells in the gall bladder epithelium of the mouse

Summary Electron microscopic observations have been made of the two epithelial cell types, light barrel-shaped and dark rod-shaped cells in the gall bladder of the mouse.The light cells have a voluminous cytoplasm of low electron opacity in which cell organelles such as mitochondria, elements of granular endoplasmic reticulum, and free ribosomes undergo more or less degenerative changes. However, there are a relatively abundant Golgi apparatus and numerous lysosomal dense bodies. The ultrastructural features of the light cells suggest that they are an aged, degenerative cell type with declining functional activity and a high degree of hydration.The dark cells are characterized by a high concentration of mitochondria and free ribosomes, more or less distinctive elements of granular endoplasmic reticulum, and well developed components of the Golgi apparatus. Such ultrastructural characteristics indicate that the dark cell type has a high synthetic activity.What has been observed in the present study can well be correlated with the results of previous studies on the same cells by methods of light microscopic histochemistry.     

Effect of actinomycin D on the ultrastructure of the hippocampal cortex

Ultrastructure of the CA1 zone in the rat dorsal hippocampus has been investigated after injection of actinomycin D into the cerebral lateral ventricles. Actinomycin D possesses a wider spectrum of action as it was previously thought. The data obtained make it possible to suppose that certain cerebral disturbances (in particular, memory), produced with actinomycin D, can be dependent on or stipulated by: decreased DNA synthesis in the neuronal nuclei, disorder of RNA synthesis in neurons and astrocytes, damage of the protein synthesis apparatus only in neurons with a dense granular endoplasmic reticulum, decreasing contents of functionally active neurons, possessing a loose granular endoplasmic reticulum, decreasing production of energy by mitochondria of synapses, neurons and astrocytes.     

Fine structure of the corpus Luteum of the raccoon during pregnancy

Summary Ultrastructure of the granulosa lutein cells of the raccoon from throughout pregnancy has been described. The lutein cells often from epithelial cords which are separated by the connective tissues, capillaries and lymphatics. Based on the arrangements and modifications of the cytoplasmic organelles and inclusions, three types of lutein cells have been recognized. The type I lutein cells predominantly contain tubular, agranular endoplasmic reticulum, juxtanuclear Golgi complexes, a few round to rod-shaped mitochondria, some free ribosomes, and occasional lipid droplets. Occasionally the tubular cristae of mitochondria and tubular smooth endoplasmic reticulum appear contiguous. The type II cells contain abundant lace-like and/or stacked fenestrated endoplasmic reticulum cisternae that frequently form membranous whorls, some tubular, agranular endoplasmic reticulum, mitochondria, and lipid droplets. Mitochondria are usually small, but unusual large ones also occur. The small, rod-to round-shaped mitochondria usually have tubular cristae; but the large, oval, elongate, and cup shaped mitochondria possess tubular, lamellar, plate like, and whorl-like cristae. The plasma membranes of the cells are complexly elaborated and folded, especially when apposing each other. In favorable sections, strands of fenestrated cisternae appose the folds of the plasma membranes. In general, the amount of cytoplasmic organelles and inclusions vary greatly in the cells. The type III cells predominantly contain lipid droplets and sparse cytoplasmic organelles. The type I and II cells are found throughout pregnancy, but the type III cells are observed from mid gestation to term. The cytological features of type I and II cells suggest that they probably secrete most of the steroids, whereas the type III cells primarily store lipids.This research was supported by UPSHS grant AM-11376 and NIH contract 69-2136.     

Alkalinization-Induced Changes in Intracellular Calcium in Rat Spinal Cord Neurons

It is well-known that pH changes can influence a lot of cellular processes. In this work, we have specifically studied the influence of alkalinization, which can be developed in spinal cord neurons during hyperventilation (respiratory alkalosis) and chronic renal failure (metabolic alkalosis) on calcium homeostasis. Application of Tyrode solution with increased pH (pH = 8.8) to secondary sensory neurons isolated from rat spinal dorsal horn induced elevation of intracellular free calcium concentration in the cytosol ([Ca2+]i) if applied after membrane depolarization. Repetitive application of alkaline solution led to disappearance of such elevations. Depletion of endoplasmic reticulum (ER) calcium stores by 30 mM caffeine almost completely blocked the effect of elevated extracellular pH. If caffeine-induced [Ca2+]i transients were evoked during alkalinization, their amplitudes were decreased by 41%. Preapplication of 500 nM ionomycin resulted in disappearance of alkalinization-induced [Ca2+]i transients, whereas prolonged applications (for 20 min) of 200 nM thapsigargin, a blocker of Ca2+ ATPase of the endoplasmic reticulum, resulted in disappearance of the rapid phase of the [Ca2+]i transients induced by alkalinization. Preapplication of the mitochondrial protonophore CCCP (10 microM) also induced changes in the alkalinization-induced calcium response--it lost its peak and was transformed into an irregular wave terminating in several seconds. The data obtained indicate that alkalinization induces an increase of [Ca2+]i level in the investigated neurons via a combined action of both intracellular Ca2+-accumulating structures--the endoplasmic reticulum and mitochondria. This suggestion was supported by morphological data that both structures in these neurons are tightly connected and may interact during release of accumulated calcium ions.     

The role of components of the endoplasmic reticulum in the biosynthesis of cytochrome P-450.

1. Antibodies have been prepared to rat hepatic cytochrome P-450 and their specificity demonstrated. These antibodies have been used to investigate the biosynthesis of cytochrome P-450 in vitro and in situ in various components of the endoplasmic reticulum. 2. A preparation of heavy rough endoplasmic reticulum translocates proteins newly biosynthesized in vitro vectorially into the luminal space and these are released by low concentrations of deoxycholate. A significant proportion of the radioactivity found in this released fraction is incorporated into cytochrome P-450. 3. Following incorporation of [14C]leucine by perfused rat liver, radioactively labelled cytochrome P-450 can be found in the intrascisternal content of heavy rough, light rough and smooth endopalsmic reticulum and also in a solublized Golgi preparation. 4. We suggest that at least part of the newly biosynthesized cytochrome P-450 is translocated into the intracisternal space of the rough endoplasmic and then passes through the other components of the endoplasmic reticulum before insertion at its ultimate membrane locus.     

Novel Robinow syndrome causing mutations in the proximal region of the frizzled-like domain of ROR2 are retained in the endoplasmic reticulum

ROR2 is a member of the cell surface receptor tyrosine kinase (RTKs) family of proteins and is involved in the developmental morphogenesis of the skeletal, cardiovascular and genital systems. Mutations in ROR2 have been shown to cause two distinct human disorders, autosomal recessive Robinow syndrome and dominantly inherited Brachydactyly type B. The recessive form of Robinow syndrome is a disorder caused by loss-of-function mutations whereas Brachydactyly type B is a dominant disease and is presumably caused by gain-of-function mutations in the same gene. We have previously established that all the missense mutations causing Robinow syndrome in ROR2 are retained in the endoplasmic reticulum and therefore concluded that their loss of function is due to a defect in their intracellular trafficking. These mutations were in the distal portion of the frizzled-like cysteine rich domain and kringle domain. Here we report the identification of two novel mutations in the frizzled-like cysteine-rich domain of ROR2 causing Robinow syndrome. We establish the retention of the mutated proteins in the endoplasmic reticulum of HeLa cells and therefore failure to reach the plasma membrane. The clustering of Robinow-causing mutations in the extracellular frizzled-like cysteine-rich domain of ROR2 suggests a stringent requirement for the correct folding of this domain prior to export of ROR2 from the endoplasmic reticulum to the plasma membrane. GenBank accession number ROR2, M97639.     

Ultrastructural changes in the MP3 neuron of the mollusk Lymnaea stagnalis after cryopreservation of the isolated brain

Investigation of a possibility of long-term storage of frozen (-196 degrees C) viable neurons and nervous tissue is one of the central present day problems. In this study ultrastructural changes in neurons of frozen-thawed snail brain were examined as a function of time. We studied the influence of cryopreservation, cryoprotectant (Me2SO), cooling to 4-6 degrees C, and a prolonged incubation in physiological solution at 4-6 degrees C on dictyosomes of Golgi apparatus, endoplasmic reticulum (ER) cisternae and mitochondria. It has been found that responses of these intracellular structures of cryopreserved neurons to the above influences are similar: dissociation of Golgi dictyosomes, swelling of endoplasmic reticulum cisternae and mitochondrial cristae. Both freezing-thawing and cryoprotectant were seen to cause an increase in the number of lysosomes, liposomes, myelin-like structures, and to form large vacuoles. The structural changes in molluscan neurons caused by cryopreservation with Me2SO (2 M) were reversible.     

THE RESPONSE OF VENTRAL HORN NEURONS TO AXONAL TRANSECTION

The morphological changes induced in the frog ventral horn neurons by axonal transection have been studied with the electron microscope. During the first 2 wk after axotomy the neuronal nucleus becomes more translucent and the nucleolus becomes enlarged and less compact. The cisternae of the granular endoplasmic reticulum vesiculate and ribosomes dissociate from membranes. Free ribosomes and polysomes are dispersed in the cytoplasmic matrix. Neurofilaments and neurotubules are increased in number. These structures appear to be important in the regeneration of the axon. It is proposed that neurotubules, neurofilaments, and axoplasmic matrix are synthesized by the free polyribosomes in the chromatolytic neuron. By the fourth postoperative week, the neurons show evidence of recovery. The cytoplasm is filled with profiles of granular endoplasmic reticulum and many intercisternal polysomes. The substances being manufactured by the newly formed granular endoplasmic reticulum are not clearly defined, but probably include elements essential to electrical and chemical conduction of impulses. The significance of these observations in respect to recent studies of axoplasmic flow is discussed.