Morphology, ultrastructure and synapse distribution of putative serotonergic salivary neurons in the locust

Neurons innervating the locust salivary glands have been investigated using the electron microscope. We have examined the projections in the suboesophageal ganglion of two identified salivary neurons by differential double labelling with cobalt ions, followed by silver intensification. Numerous synaptic inputs occur on the arborizations of the salivary neurons, particularly on fine branches and on small spines arising from larger branches. Although a few instances occur of common input to both salivary neurons from a single presynaptic element, many of the appositions between branches seen in the light microscope do not represent functional connections. A few structures resembling presynaptic dense bodies have been observed in salivary neuron profiles, but with few synaptic vesicles. Large dense granules are present in some labelled profiles, but not in the vicinity of synapse-like membrane specializations. We have also examined unidentified neuron profiles within the acini of the salivary glands, which contain large dense granules and small vesicles. There is good evidence that these unidentified terminals correspond to the suboesophageal salivary neurons. The central arborizations of the salivary neurons appear to serve largely as sites of synaptic input, whereas the peripheral terminals are likely sites of transmitter release. The results are considered in the context of the known immunoreactivity of the salivary neurons with antibodies to 5-hydroxytryptamine, and compared with analogous systems in other insect groups.     

On certain fluorescent axon terminals containing granular synaptic vesicles in the cerebellar nucleus lateralis

Summary Thin, unmyelinated, green fluorescent fibers bearing fine beads or varicosities have been found in the neuropil and near blood vessels of the nucleus lateralis. In electron micrographs these fibers are identifiable as a class of axons, the CAT fibers, which contain large and small granular synaptic vesicles and agranular vesicles in their varicosities. There are two types of CAT fiber. 1) The CAT₁ terminals contain many large and elongated vesicles, 700–1700 Å in size, with dark, homogeneously dense centers; a few small granular vesicles each with an intensely osmiophilic particle, and small agranular vesicles. These terminals have not been seen in synaptic contact with other elements of the neuropil. 2) The CAT₂ terminals have a very thin unmyelinated connecting thread between small varicosities. The varicosities contain small agranular synaptic vesicles and small granular ones containing either a single dense particle, or an elliptical, intensely osmiophilic droplet flanked by lighter semicircular particles. Large granular vesicles, 750–950 Å each, with a variably dense center, are also found. These terminals form conventional axodendritic synapses with Gray's type 1 synaptic junctions and the subsynaptic specialization of Taxi, as well as synapses on thorns of spiny neurons.It is suggested that the CAT₁ and CAT₂ fibers may be the electron microscope equivalents of norepinephrine- and 5-hydroxytryptamine-containing fluorescent axons. These fibers probably have extrinsic origins since no fluorescent cells or perikarya with small or large granular vesicles have been found in the lateral nucleus. Their origins, however, are unknown. The proximity of these fluorescent fibers to blood vessels is discussed, and their functions are the subject of some speculation.Supported in part by U.S. Public Health Service grants NS10536, NS03659, Training grant NS05591 from the National Institute of Neurological Diseases and Stroke and a William F, Milton Fund Award from Harvard University.     

The dense core of vertebrate central nervous system synapses revealed by potassium permanganate fixation as formed by small membrane-bounded vesicles

Large dense core vesicles occur in small number in the company of clear synaptic vesicles in various parts of the central nervous system following osmium or aldehydes fixation. When fixed by potassium permanganate large vesicles occur that contain one or more membrane-bounded small spherical vesicles, as is demonstrated in the habenular nuclei of the frog Rana esculenta. On a morphological basis we suppose that the population of this kind of synapse consists of the same type of vesicles, the majority being free while some are enclosed within a larger vesicle. An hypothesis on the meaning of these composite vesicles is presented.     

The ultrastructure of the olivary pretectal nucleus in rats. A tracing and GABA immunohistochemical study

 The olivary pretectal nucleus is a primary visual centre, involved in the pupillary light reflex. In the present study an ultrastructural analysis was made of the olivary pretectal nucleus by means of separate, anterograde and retrograde tracing techniques and immunohistochemistry of gamma-aminobutyric acid. Large-projection neurons and two types of gamma-aminobutyric acid-immunoreactive (GABA-ir) neurons are observed in the olivary pretectal nucleus. The primary dendrites of the projection neurons have a dichotomous appearance, the secondary dendrites a multipolar appearance. At the ultrastructural level the projection neurons have well-developed Golgi fields, abundant rough endoplasmic reticulum and the nucleus is always heavily indented. Numerous small GABA-ir neurons and a few medium-sized GABA-ir neurons are found. The small GABA-ir neurons contain a few stacks of rough endoplasmic reticulum and the nucleus is oval-shaped. The medium-sized GABA-ir neurons have well-developed Golgi fields, a moderate number of rough endoplasmic reticulum stacks and an indented nucleus. GABA-positive dendritic profiles containing vesicles also are observed. In the neuropil of the olivary pretectal nucleus, retinal terminals are found that contain round clear vesicles and electron-lucent mitochondria. They make asymmetric synaptic contacts (Gray type I) with dendritic profiles and with profiles containing vesicles. Terminals originating from the contralateral olivary pretectal nucleus exhibit small, round clear vesicles, electron-dense mitochondria and make asymmetric synaptic contacts (Gray type I) mainly with dendritic profiles. Two types of GABA-ir terminals were found. One type is incorporated in glomerulus-like arrangements, whereas the other type is not. GABA-ir terminals contain pleomorphic vesicles, electron-dense mitochondria and make symmetric synaptic contacts (Gray type II). Retinal terminals, terminals originating from the contralateral olivary pretectal nucleus and GABA-ir terminals are organized in glomerulus-like structures, in which dendrites of the large projection neurons form the central elements. Triadic arrangements are observed in these structures; a retinal terminal contacts a dendrite and a GABA-ir terminal and the GABA-ir terminal also contacts the dendrite. The complexity of the synaptic organization and the abundancy of inhibitory elements in the olivary pretectal nucleus suggest that the olivary pretectal nucleus is strongly involved in processing visual information in the pupillary light reflex arc. Received: 17 July 1996 / Accepted: 24 September 1996     

Exocytosis from large dense cored vesicles outside the active synaptic zones of terminals within the trigeminal subnucleus caudalis: a possible mechanism for neuropeptide release

It has been hypothesized that chemical interactions between neurons in the central nervous system can occur in the absence of well defined synaptic complexes, but morphological correlates have been difficult to find. The present study demonstrates exocytotic release from large (70-130 nm) dense cored vesicles at structurally nonspecialized areas along the plasmalemma of structurally different categories of terminals and occasionally from dendrites and axons within the neuropil of the trigeminal subnucleus caudalis. In rats, the marginal (lamina I) and substantia gelatinosa (lamina II) layers contain the central terminals of primary afferent fibers from the infraorbital nerve that supply the skin and whiskers (vibrissae). Different types of interneurons are also present and may modify the input being relayed to higher centers. While exocytotic profiles were present in control animals, they increased significantly (P less than 0.01) on the ipsilateral side 1-24 h after a unilateral skin lesion in the vibrissae area. A second increase (P less than 0.001) occurred 14-15 days after the lesion. Virtually all examples of large vesicle exocytosis were observed at structurally nonspecialized sites while those at the active synaptic zones involved small clear vesicles. Substance P-like immunofluorescence, present in controls and on the ipsilateral side during the first 6 days, subsequently declined until 4 weeks after surgery when some recovery was noted. The increase in large vesicle exocytosis and the decrease in substance P are interpreted to reflect functional adjustments of different neurons in response to the lesion. The exocytosis involving large dense cored vesicles may serve to deliver transmitters and/or neuropeptide modulators to appropriate receptors in a wider area than release into a specialized synaptic cleft would allow.     

An ultrastructural study of 5-hydroxytryptamine-, thyrotropin-releasing hormone- and substance P-immunoreactive axonal boutons in the motor nucleus of spinal cord segments L7-S1 in the adult cat

The distribution and fine structure of 5-hydroxytryptamine-, thyrotropin-releasing hormone- and substance P-immunoreactive synaptic boutons and varicosities were studied in the motor nucleus of the spinal cord segments L7-S1 in the cat, using the peroxidase-antiperoxidase immunohistochemical technique and analysis of ultrathin serial sections. The 5-hydroxytryptamine-, thyrotropin-releasing hormone- and substance P-immunoreactive boutons had a similar ultrastructural appearance as judged from serial section analysis. The boutons could be classified into two types on the basis of their vesicular content, with one type containing a large number of small agranular vesicles together with only a few, if any large granular vesicles, while the other type contained a large number of large granular vesicles in addition to small agranular vesicles. The vesicles were spherical or spherical-to-pleomorphic. Postsynaptic dense bodies (Taxi bodies) were occasionally observed in relation to all three types of immunoreactive boutons, which almost invariably formed synaptic junctions with dendrites. Judged by the calibre of the postsynaptic dendrites, the boutons were preferentially distributed to the proximal dendritic domains of motoneurons. In one case, a substance P-immunoreactive bouton formed an axosomatic synaptic contact. In addition to synaptic boutons, 5-hydroxytryptamine-, thyrotropin-releasing hormone- and substance P-immunoreactive axonal varicosities containing a large number of large granular and small agranular vesicles but lacking any form of conventional synaptic contact were observed. Such varicosities were either directly apposing surrounding neuronal elements or separated from the neurons by thin glial processes. The origin of the immunoreactive boutons was not traced, but it was thought likely that the main source of the boutons was neurons with their cell bodies located in the medullary raphe nuclei.     

Ultrastructural organization of the interstitial subnucleus of the nucleus of the tractus solitarius in the cat: Identification of vagal afferents

Summary This electron microscopic study, based on serial section analysis, describes the synaptic organization of the interstitial subnucleus of the nucleus of the solitary tract and identifies the terminals of the vagal primary afferents utilizing degeneration and HRP transport. The interstitial subnucleus contains sparsely scattered cell bodies, numerous dendrites and axon terminals, and bundles of unmyelinated and myelinated axons. The cell bodies which are small in diameter have an organelle poor cytoplasm and a large invaginated nucleus.Axon terminals can be classified into two main types according to their vesicular shape. The first type contains clear, round vesicles and can be further subdivided into two subgroups on the basis of their morphology and the size of their vesicles. In the first subgroup the terminals are small, contain a few mitochondria and their vesicles are densely packed with an homogeneous size. In the second subgroup the terminals which vary from small to large, contain many mitochondria and contain round vesicles which are heterogeneous in size. The second main terminal type consists of axon terminals containing pleomorphic vesicles which are associated with asymmetrical or symmetrical synaptic contacts on dendrites. Axo-axonic contacts are present in the interstitial subnucleus. In general, the presynaptic axon terminals contain pleomorphic vesicles and the postsynaptic elements contain round vesicles of varying size. In some dendrites, identified by the presence of ribosomes, groups of round and/or pleomorphic vesicles are found associated with synaptic contacts. These dendrites are presynaptic to conventional dendrites and postsynaptic to axon terminals. After removal of the nodose ganglion, degenerative alterations are seen only at the caudal and middle levels of the interstitial subnucleus. Degeneration occurs in a few myelinated axons and in axon terminals which usually contain a mixture of small and larger round, clear vesicles. After HRP injection into the vagus nerve, the HRP reaction product is visible in axon terminals filled with clear, round vesicles which are heterogeneous in size. The labelled axon terminals establish single or multiple synaptic contacts.This study demonstrates that terminals of vagal primary afferents consist principally of terminals of the second subgroup. The morphology of these terminals are compared to primary afferents in the brainstem and spinal cord.     

Fine structure of nerves in the regenerating limb of the newt Triturus

The structural integrity of some tissues and the regeneration of extremities in some vertebrates depends upon the nervous system. To investigate the structure of nerves exhibiting trophic function, nerves in regenerating forelimbs of adult newts, Triturus viridescens, were studied with the electron microscope. Nerve fibers sprout from the transected axons, 2–3 days after amputation of the limb, and invade all portions of the blastema and epidermis in large numbers. Regenerating nerve fibers contain a greatly increased amount of smooth-surfaced channels of endoplasmic reticulum containing moderately dense material, an increased number of microtubules, and large (1000 Å) membrane-bounded dense granules. The latter were not observed normally and could be distinguished from synaptic vesicles and dense-core vesicles thought to contain catechol amines. In larger nerves, the organelles are distributed in the peripheral axoplasm around a central zone containing neurofilaments. The relationship of the fine structural changes occurring during regeneration to the trophic action of nerves and the regeneration of nerve fibers is discussed. The tortuous membranous tubules of endoplasmic reticulum could serve as channels for the transport of substances, either trophic material or materials necessary for growth of the nerve, down the axon. Microtubules may play a role in the regulation of form of the growing axon and also be related to axoplasmic flow or migration of particles (e.g., granules) along their length. The large membrane-bounded dense granules appearing during regeneration resemble neurosecretory granules, which have been associated with regeneration in some invertebrates. These structures could, therefore, contain a trophic substance or hormone that is transported down the axon, released into the intercellular spaces, and controls subsequent regeneration of the limb.     

Argentaffin and other “endocrine” cells of the small intestine in the adult mouse. I. Ultrastructure and classification

Electron microscopic examination of the epithelium of crypts and villi in the jejunum of the adult mouse revealed the existence of three main types of cells which may be classified within the group of argentaffin and other “endocrine” cells of the gastrointestinal tract. Sixty per cent of the cells are characterized by the presence of biconcave granules. They are referred to as biconcave granule cells (Type EC). While most granules are biconcave, some of them are spherical and may show a particulate structure. Those cells with 95% or more biconcave granules predominate in the crypt, while those with 50–75% predominate in villi. The cells of this type are considered by some authors to be true argentaffin or enterochromaffin cells. Thirty-seven per cent of the cells have spherical dense granules, a few of which look like empty vesicles. However, because of the marked differences between the sizes of the granules, these cells are classified into two categories. Those with granules averaging 360 mμ in diameter are referred to as large spherical granule cells (Type L, 13%), while those with granules averaging 250 mμ in diameter, are referred to as small spherical granule cells (Type S, 24%). Both types are found in crypts and villi. Three per cent of the cells do not fit into these categories. All these uncommon cells contain spherical granules; and, in addition, some have oval ones; others show large halos; and still others include granules of different sizes and densities. The existence of several “endocrine” cell types suggests that they play different roles in the jejunum of the mouse.     

On the identification of the afferent axon terminals in the nucleus lateralis of the cerebellum an electron microscope study

Summary Axon terminals in the neuropil of the lateral nucleus can be divided into six classes, each with a specific constellation of characteristics that consistently occur together. Two of these classes have synaptic varicosities with elliptical synaptic vesicles, one in a dense, the other in a sparse matrix, and both make axosomatic and axodendritic synapses. The remaining four classes all have round synaptic vesicles and do not make axosomatic synapses. In the first of these four, the vesicles are tightly packed in a dense matrix, in another they are loosely dispersed, and in the third they are clustered. In the fourth, large granular vesicles predominate. Of these six classes, the most numerous belong to the axons of the Purkinje cell terminal arborization. These boutons resemble their counterparts in the cerebellar cortex, the recurrent collaterals of the Purkinje axon. They have elliptical and flat synaptic vesicles in a dark matrix. The varicosities terminate on somata and dendrites of large and small neurons and constitute the majority of their input. Purkinje axons constitute 86% of the total population of terminals on large neuronal perikarya and 50% of those on their dendrites, but only 78% on the somata of small neurons and 31% on their dendrites. The terminals of climbing fiber collaterals are recognized by their resemblance in electron micrographs to the terminals of the climbing fiber arborization in the cerebellar cortex. They bear round synaptic vesicles packed into a dense axoplasmic matrix and make Gray's type 1 axodendritic synapses with large and small neurons. These axons are restricted to the lateral and ventral aspects of the nucleus and constitute 5% of the terminals on large cell dendrites and 6% of those on small neurons. The axons tentatively identified as collaterals of mossy fibers are myelinated fibers with a light axoplasm containing round synaptic vesicles, dispersed throughout their varicosities. They make Gray's type 1 synapses and constitute a fair percentage of the total axodendritic contacts in the neuropil, 22% on large neurons and 28% on small neurons. The bases for these tentative identifications are discussed in detail, as are the various synaptic relationships undertaken by each class of axon. The remaining 4 classes of axons of the neuropil will be described in subsequent papers.Supported in part by U.S. Public Health Service grants NS 10536 and NS 03659, Training grant NS 05591 from the National Institute of Neurological Diseases and Stroke, and a William F. Milton Fund Award from Harvard University.     

Synaptic circuitry in the retinorecipient layers of the optic tectum of the lamprey (Lampetra fluviatilis). A combined hodological, GABA and glutamate immunocytochemical study

The ultrastructure of the retinorecipient layers of the lamprey optic tectum was analysed using tract tracing techniques combined with GABA and glutamate immunocytochemistry. Two types of neurons were identified; a population of large GABA-immunonegative cells, and a population of smaller, highly GABA-immunoreactive interneurons, some of whose dendrites contain synaptic vesicles (DCSV). Five types of axon terminals were identified and divided into two major categories. The first of these are GABA-immunonegative, highly glutamate-immunoreactive, contain round synaptic vesicles, make asymmetrical synaptic contacts, and can in turn be divided into AT1 and AT2 terminals. The AT1 terminals are those of the retinotectal projection. The origin of the nonretinal AT2 terminals could not be determined. AT1 and AT2 terminals establish synaptic contacts with DCSV, with dendrites of the retinopetal neurons (DRN), and with conventional dendritic (D) profiles. The terminals of the second category are GABA-immunoreactive and can similarly be divided into AT3 and AT4 terminals. The AT3 terminals contain pleiomorphic synaptic vesicles and make symmetrical synaptic contacts for the most part with glutamate-immunoreactive D profiles. The AT4 terminals contain rounded synaptic vesicles and make asymmetrical synaptic contacts with DRN, with DCSV, and with D profiles. A fifth, rarely observed category of terminals (AT5) contain both clear synaptic vesicles and a large number of dense-core vesicles. Synaptic triads involving AT1, AT2 or AT4 terminals are rare. Our findings are compared to these of previous studies of the fine structure and immunochemical properties of the retinorecipient layers of the optic tectum or superior colliculus of Gnathostomes.     

Ultrastructure and synaptic relations of neural elements containing glutamic acid decarboxylase (GAD) in the perigeniculate nucleus of the cat

Summary The perigeniculate nucleus of the cat (PGN) was examined at light and electron microscopic levels after immunocytochemical labeling for the gamma-aminobutyric acid (GABA) synthesizing enzyme, glutamic acid decarboxylase (GAD). In light microscopic sections, virtually all perikarya were found to be labeled (GAD+), as well as proximal dendrites, fibres and punctiform elements. Cells in the thalamic reticular nucleus (TRN) dorsal to PGN were also labeled. Ultrastructural analysis of PGN showed immunoreactivity in all somata, in dendrites and in the following vesicle containing profiles: 1.) F1 terminals, which are characterized by large size, dark mitochondria, and pleomorphic vesicles. These terminals form symmetrical synaptic contacts with somata, somatic spines and with dendrites of GAD+ PGN cells. 2.) F2 terminals, which are smaller than F1 terminals, contain also pleomorphic vesicles and frequently make serial synapses of the symmetric type with other F2 terminals. Presumably, F1 terminals are formed by collaterals of PGN-cell axons and F2 terminals by vesicle containing dendrites of PGN cells. Terminals devoid of immunoreactivity included: 1.) RLD terminals characterized by large size, round vesicles, dark mitochondria, and by asymmetric synaptic contacts with somata, especially with somatic spines, and with dendrites of GAD+ perigeniculate neurons; 2.) RSD terminals, characterized by small size, round vesicles and dark mitochondria, which make asymmetric synapses with GAD+ dendrites of medium and small size; 3.) Multivesicular (MV) terminals with variably shaped vesicles including dense core vesicles synapsing on GAD+ dendrites. There are reasons to believe that RSD terminals belong to corticofugal axons and RLD terminals to collateral axons of LGN relay cells. The origin of MV terminals remains to be determined. The GABAergic nature of the PGN cells conforms with the presumed function of these cells as mediators of inhibition of LGN relay cells. The complex synaptic relations observed between GAD+ elements in the PGN would allow for reciprocal inhibition between perigeniculate cells.Supported in part by NIH grants EY02877 to V.M. Montero and HD 03352 to the Waisman Center     

Differential expression and subcellular localization of secretogranin II and synaptophysin during early development of mouse hypothalamic neurons in culture.

Mature neurons contain two distinct regulated secretory pathways, characterized electron microscopically by so-called large dense core vesicles and small synaptic vesicles, respectively. Each vesicle type is characterized by vesicle-specific proteins, such as the granins (chromogranins/secretogranins) for the matrix of large dense core vesicles and synaptophysin for the membrane of small synaptic vesicles. So far, no data exist on the biogenesis of these two distinct vesicle types during neuronal development. We have used secretogranin II and synaptophysin as markers for the biogenesis of these two vesicle types during the development of mouse hypothalamic neurons in culture, using immunocytochemistry and biochemical analyses. By immunofluorescence, we found that secretogranin II appears as early as synaptophysin, but in a subset of neurons only, and with different subcellular localizations. It was observed in cytoplasmic areas where little or no synaptophysin immunofluorescence was detected, such as lamellipodia, emerging neurites and growth cones. At later stages, the proportion of secretogranin II-containing varicosities remained steady whereas that of synaptophysin-containing varicosities increased dramatically. By quantitative analysis we found that the level of expression of synaptophysin increased several-fold during synaptogenesis whereas that of secretogranin II decreased. These data suggest that large dense core vesicles and small synaptic vesicles can be formed separately and expressed at different levels. They provide evidence for a differential biogenesis of these two distinct vesicle types.     

Ultrastructural evidence for an innervation of epithelial enterochromaffine cells in the guinea pig duodenum

The innervation of the duodenal enterochromaffine cells (E.C.) of the guinea pig was studied at the electronmicroscopic level. Pretreatment with 5-OH-dopamine was performed to visualize catecholaminergic (CA) nervous elements. Near the basement membrane of all examined E.C. in the crypts, bundles of unmyelinated nerve processes were observed, only partly ensheathed in a Schwann cell cover. At least 4 types of processes could be observed. 1) Boutons containing only small clear vesicles, probably cholinergic fibres; 2) boutons with small clear vesicles, and in addition large (200 nm) granules with a dense matrix (P-type-fibres); 3) boutons with small electron-dense vesicles, probably CA-fibres; and 4) processes with few vesicles but having the appearance of dendrites. No typical synaptic arrangements were observed, but the minimal distance between the E.C. and the nerve bundles was 150 to 250 nm, thus well within the functional limits of the “autonomic gap”. Thus, epithelial E.C. may be influenced by several types of nervous elements, including CA-fibres.     

大鼠皮质大颗粒泡的超微结构

用电镜在4只猫听区皮质和3只大白鼠额叶皮质中观察了大颗粒泡的超微结构。在2031个突触面上,共见到大颗粒泡282个。1.大颗粒泡的位置:在104个突触前囊中含有大颗粒泡135个,它们多位于突触前囊的后部(64/135),位于前囊的中部或接近突触前膜者较少。有的大颗粒泡不在突触前囊内,而位于轴突、树突或个别例在树突侧棘中。2.大颗粒泡数目:大脑皮质中的大颗粒泡数目较少。在各种成分中(包括突触前囊、轴突、树突以及其他成分)计见282个大颗粒泡,在一个成分中含1个大颗粒泡的占多数(78.80％)。个别例有含有6个者。多数突触前囊无大颗粒泡。3.大颗粒泡的形态:多为圆形或卵圆形,亚铃形或不整形者也有之。致密核心多数致密,有的并不致密而显得灰淡,有的致密核心呈网状,或在一个包膜内有两个核心。有的膜下环并不明显。4.大颗粒泡的大小不一。本文对大颗粒泡和某些递质的可能关系进行了讨论。     

Chondroitin sulfate and electron lucent bodies in the pericellular rim about unshrunken hypertrophied chondrocytes of chick long bone.

Direct observation of unstained, 1 mm thick blocks of fresh epiphyseal cartilage from tibia of 15- and 18-day-old chick embryos revealed shrunken chondrocytes on its cut surfaces but unshrunken chondrocytes deep within the tissue blocks. The unshrunken hypertrophied chondrocytes are rimmed with refractile substance identified as chondroitin sulfate removable with hyaluronidase. This substance is stained metachromatically red with toluidine blue, and is stained with ruthenium red and with ruthenium red-OsO4. The latter, observed with the electron microscope, is present as an electron dense rim, specifically about the unshrunken, hypertrophied chondrocytes between the plasma membrane and lacunar wall. By rendering the chondroitin sulfate electron dense with RR-OsO4, electron lucent bodies (ELB) were revealed specifically about the hypertrophied chondrocytes. The ELB contain an electron dense core with radiating fibrils. The content and source of ELB, also found in the intercellular matrix, are not known. The 0.1% toluidine blue solution containing 0.2 M MgC12 or 0.4% NaCl or KCl stained juxtanuclear clusters of granules metachromatically red. The location of intracellular granules was believed to represent a cluster of Golgi-derived vesicles. The pericellular metachromatic, RR-OsO4-positive rim is believed to be an accumulation of externalized juxtanuclear metachromatic granules. The possibility that the ELB may also be externalized content of Golgi vesicles was entertained.     

The synaptic organization of terminals traced from individual labeled retino-geniculate axons in the cat

Individual retino-geniculate axons in the dorsal lateral geniculate nucleus of the cat were filled with horseradish peroxidase and studied with both the light and electron microscope. A procedure was followed which allowed us to identify the size, shape and arrangement of particular terminal swellings by light microscopy and then to study their patterns of synaptic contacts with the electron microscope. Many of the labeled terminals in laminae A and Al have the same fine structural features as retino-geniculate terminals that have been described previously. They are large, contain round synaptic vesicles and pale mitochondria, and are the central processes in glomeruli where they form asymmetric contacts with dendrites and terminals containing pleomorphic synaptic vesicles. Other terminals have the same cytological features but are quite small and are not the central processes in glomeruli. Some of these small terminals form simple axo-dendritec contacts while others participate in very large glomeruli containing several terminals from a single retino-geniculate axon. These different patterns of synaptic contacts made by different terminals can be found on branches of a single axon and correspond to the variations in terminal arbors described in the preceding paper (MASON & ROBSON, 1978).     

Amphicrine carcinoma of breast with giant granules: an immunohistochemical, histochemical and ultrastructural study

A moderately differentiated grade 2 invasive ductal carcinoma was diagnosed in the right breast of an 81-year-old woman. The uniform nuclear profiles and moderately abundant granular cytoplasm suggested a neuroendocrine tumour and a Grimelius stain was positive. Neurone specific enolase, synaptophysin and somatostatin stained positively, and casein was interpreted as positive although with some background staining. By electron microscopy, tumour cells possessed desmosomes, tonofibrils, intercellular lumina, lamina and dense granules. Rounded dense granules 160-480 nm in diameter resembled neuroendocrine granules. They were found in both luminal and basal areas. Fewer and much larger ('giant') granules had a rounded profile and were up to 5 microm across. The smaller cytoplasmic granules were mostly Grimelius-positive while giant granules were negative. The smaller granules were also uranaffin-positive, but no uranaffin-positive cytoplasmic giant granules were encountered. Both small and giant granules were observed in lumina, and here both were uranaffin-positive. Intraluminal giant granules had a substructure of small pale lipid-like lacunae, and some had irregular profiles. The exceptional size of these exocrine granules is emphasised, and the nature of both the small and giant granules discussed in this amphicrine carcinoma.     

Two types of thyrotropes (TSH cells) in the adenohypophysis of the untreated mouse

Employing immunohistochemical techniques with OsO4 single fixation, we identified mouse pituitary thyrotropes (TSH cells). The cells stained with TSH antiserum are angular or slender with a small nucleus. These cells contain small secretory granules (about 120-200 nm in diameter) and numbers of cored vesicles, either attached to the cell membranes or lying free in a relatively electron transparent cytoplasm. In the untreated male mouse, the TSH cell modifies its morphology according to the functional phase. The first form of TSH cells [Type I] is small. The endoplasmic reticulum and the Golgi apparatus are not prominent, and secretory granules are very few. In the second hypertrophied form [Type II], the endoplasmic reticulum is very prominent and occurs as a series of grossly dilated sacs of irregular shape. The Golgi apparatus is greatly enlarged and a large number of electron-dense secretory granules and cored vesicles are observed. Type II thyrotropes are rarely encountered in normal tissue.