The lack of a structured blood-brain barrier in the onychophoran Peripatus acacioi

Summary Onychophorans are ‘living fossils’ frequently purported to have evolved from the same ancestor as the arthropods and annelids. In the CNS ofPeripatus acacioi, beneath an outer acellular neural lamella, glial cells ensheath the cerebral ganglion and the nerve cords. These glial cells are, however, attenuated and rather few in number and, although they interdigitate with one another, they seem to lack intercellular junctions. Exogenous tracers penetrate between them and into the underlying neuropile, suggesting that there is no structural blood-brain barrier. Throughout the nervous tissue, extracellular spaces occur which contain banded collagen fibrils embedded in a matrix material. Thin glial cell processes, characterized by dense filaments, surround these regions and frequently form hemi-desmosomes with the extracellular matrix. The peripheral nerve cell bodies have a range of diameters; some have the characteristics of neurosecretory neurons. Granules in such neurons are produced by the Golgi saccules and associated fenestrated membranes which also possess many coated vesicles. Comparable granules are also found in axonal tracts, but no distinct peripheral neurohaemal areas have been found. Lysosomes are common in the nerve cell bodies and are frequently in the form of multivesicular bodies or large phagocytic vacuoles. Beneath the outer nerve cells lie many tracheae, arranged as a ring around the central neuropile which consists of glial processes, extracellular matrix, axons and nerve terminals. These nerve terminals occur throughout the central neuropile and are characterized by dense pyramidal presynaptic specializations and postsynaptic subsurface cisternae. The nervous system ofPeripatus is relatively simple in its organization, in the lack of glial intercellular junctions and in the ready accessibility of substances from the external milieu.     

An ultrastructural study of neurons and non-neuronal cells in the myenteric plexus of the rabbit colon

The myenteric plexus of the rabbit colon showed a degree of structural organization that was unusually high for the peripheral nervous system, providing a basis for the complex integrative activity which is known to occur. It resembled central nervous tissue in several respects: a wide range of neuron types was present; the proportion of glial cells to neurons was about 2:1; and there was a densely packed, avascular neuropil, not penetrated by connective tissue. Most neurons had at least one surface exposed to the extraganglionic space. Clear evidence was obtained for spontaneous neuronal degeneration. Three types of non-neuronal (glial) cells were observed: Type 1, which was most common, contained many 10 nm ‘gliofilaments’ and resembled enteric glial cells or astrocytes in the central nervous system; Type 2, composing about 5% of the glial cells, had few filaments; Type 3 was seen only rarely, had a small dark nucleus, little cytoplasm, may have been of extraganglionic origin and resembled microglia of the central nervous system. Fibroblast-like cells were also present in extraganglionic sites. Schwann cells could not be identified within the myenteric ganglia.     

Synganglial morphology and neurosecretory centers of adult Amblyomma americanum (L.) (Acari: Ixodidae).

Lone star ticks, Amblyomma americanum (L.), were processed by standard histological means for paraffin embedding, sectioning, and staining by the paraldehyde-fuchsin technique. The synganglion is highly condensed around the esophagus and possesses paired optic, cheliceral, palpal, pedal I-IV nerves, and opisthosomal nerves and a single unpaired esophageal nerve. Although optic nerves were observed leading from the eyes to the protocerebrum, distinct optic ganglia were not seen in any of the preparations examined. The paraldehyde-fuchsin technique revealed 14 neurosecretory centers (11 paired, 3 unpaired) within the synganglion, which are described in relation to the underlying neuropilar structure. A previously undescribed internal subesophageal center that consists of a single cell was observed within a cluster of perikarya lying posteriorly adjacent to the esophagus. A three-dimensional reconstruction of the internal neuropilar structure of the synganglion was made, and the included neurosecretory centers were mapped. Comparisons are made to previous work on other ticks, and physiological relationships are considered.     

Evidence for dynamic interactions between pituicytes and neurosecretory axons in the rat

Several ultrastructural features of pituicytes as well as their relationship with neurosecretory axons were measured in neural lobes from rats either: non-deprived; water-deprived for 4, 12, or 24 h; or rehydrated for 12 or 24 h after an initial 24 h dehydration. A significant increase in pituicyte lipid bodies occurs with as little as 4 h of water deprivation. This is a time when several dehydration-related changes are known to occur in the somata of those neurosecretory cells sending axons to the neural lobe. Thus, increases in pituicyte lipid bodies may be related to the initiation of hormone release. In the neural lobes of water replete, non-deprived rats, pituicytes were found to contain neurosecretory axons completely enclosed by their cytoplasm. The number of these per unit area of cytoplasm decreased by 64% with 24 h of water deprivation and returned to normal levels with 24 h of rehydration. Since few of the enclosed axons were found to be degenerating, enclosure is unlikely to represent the early stages of phagocytosis.These findings suggest that during periods of little hormone secretion the neurosecretory axons are enclosed by the glial cells, but are released under conditions of increased hormone demand. This may provide the morphological framework for pituicyte involvement in inhibition and facilitation of hormone release from the neurohypophysis.     

Morphological plasticity that occurs in the neurohypophysis following activation of the magnocellular neurosecretory system can be mimicked in vitro by beta-adrenergic stimulation

The osmotic stress to rats of replacing drinking water with 2% NaCl for four days (salt loading) led to dramatic ultrastructural morphological changes in the neural lobe of the pituitary, including a decrease in the ratio of glial to neurosecretory terminal coverage of the pericapillary basal lamina. The decrease in the proportion of the basal lamina covered by pituicytes, the specialized astrocytic glial cells of the neural lobe, was due to a decrease in the number of pituicyte processes reaching the pericapillary spaces. The concomitant increase in the proportion of neuronal coverage was due to the combination of an increase in the length of individual nerve terminals and a change in the number of terminals. Similar structural changes to those seen in vivo were produced by incubation of the isolated neural lobe with the beta-adrenergic agonist isoprenaline. A decrease in the ratio of glial to neuronal coverage of the basal lamina was achieved within 1 h and could be blocked by inclusion of the beta-adrenergic antagonist propranolol. It is proposed that the morphological plasticity is explained by the active movement of pituicyte cytoplasm.     

银盾革蜱神经分泌细胞的超微结构和分泌活性的研究

用透射电镜观察了银盾革蜱（Ｄｅｒｍａｃｅｎｔｏｒｎｉｖｅｕｓ）雌虫神经分泌细胞的超微结构。神经分泌细胞位于合神经节中，具有较小的核／质比。其细胞质中含有的高尔基体、粗糙内质网以及分泌颗粒是分泌活性的标志。正常雌虫饱食后２ｄ达到分泌活性的最高峰，随后即下降，到产卵时达到最低水平。滞育雌虫饱食后亦达到分泌的高峰，并可持续至少８５ｄ，此时细胞质中出现大量的平滑内质网。     

CarcinoÏde oncocytaire bronchique

Summary We describe a bronchial tumor with a biphasic pattern. The main cellular population is composed of large cells with eosinophilic cytoplasm packed with mitochondria. These oncocytes countain few dense-core membrane limited granules. The secondary cell population has the morphological and histochemical features that characterize the typical bronchial carcinoid. A third cell type, recognizable at ultrastructural level, contains numerous neurosecretory granules and abundant large mitochondria, seeming therefore to represent a transitional cell form. We assume that oncocytes and carcinoid cells, found in the same tumor with intermediate forms, represent the extremes of a morphological spectrum. Furthermore, the oncocytic cytoplasmic components (neurosecretory granules and the mitochondria) are present in reciprocal arrangement in the carcinoid cells. These observations, together with similar reported cases suggest that the oncocytic state corresponds to a functional stage. We consider that oncocytes from this endobronchial tumor and from similar pathologic conditions reported in the literature correspond to carcinoid cells with mitochondrial hyperplasia and with diminished endocrine activity. This oncocytic carcinoid tumor is associated, in the same pulmonary lobe, with a second neoplasm that has the histological and ultrastructural features of a poorly differentiated carcinoid (oat-cell carcinoma).

Tirés-à-part: Dr. P. Walter     

Paraganglioma of the Cauda Equina: An Ultrastructural and Immunohistochemical Study of Two Cases

The ultrastructural and immunohistochemical features of 2 paragangliomas arising in the cauda equina are described. In both cases the tumor cells were arranged in small nests or cords and contained characteristic neurosecretory granules, lamellar stacks of rough endoplasmic reticulum (RER), and some well-developed Golgi apparatuses in their cytoplasm. The cells varied in electron density; the darker cells, occasionally resembling sustentacular cells, were probably dehydrated light cells because they contained a few neurosecretory granules. Sustentacular cells were difficult to identify by electron microscopy, but irregularly distributed S-100 protein and glial fibrillary acidic protein (GFAP) were found in these cells by immunostaining. Many tumor cells contained abundant neurofilaments. Curiously, a few cytokeratin-positive cells were found in 1 case. On microscopic examination, a small area of ganglioneuroma was found associated with the paraganglioma in 1 case. Ganglionic differentiation was concluded to be frequent in paragangliomas of the cauda equina region as in duodenal paragangliomas.     

Paraganglioma of the Cauda Equina: An Ultrastructural and Immunohistochemical Study of Two Cases

The ultrastructural and immunohistochemical features of 2 paragangliomas arising in the cauda equina are described. In both cases the tumor cells were arranged in small nests or cords and contained characteristic neurosecretory granules, lamellar stacks of rough endoplasmic reticulum (RER), and some well-developed Golgi apparatuses in their cytoplasm. The cells varied in electron density; the darker cells, occasionally resembling sustentacular cells, were probably dehydrated light cells because they contained a few neurosecretory granules. Sustentacular cells were difficult to identify by electron microscopy, but irregularly distributed S-100 protein and glial fibrillary acidic protein (GFAP) were found in these cells by immunostaining. Many tumor cells contained abundant neurofilaments. Curiously, a few cytokeratin-positive cells were found in 1 case. On microscopic examination, a small area of ganglioneuroma was found associated with the paraganglioma in 1 case. Ganglionic differentiation was concluded to be frequent in paragangliomas of the cauda equina region as in duodenal paragangliomas.     

Morphologic features of the caudal neurosecretory system in the blueback herring, Pomolobus aestivalis

The caudal neurosecretory system of the blueback herring, Pomolobus aestivalis, captured in freshwater, was examined. Neurosecretory cells were identified readily in terminal spinal cord segments. The axonal processes of these neurosecretory cells formed a discrete, ventrally placed bundle which terminated in a conspicuous ventral enlargement from the filum terminale. This enlargement comprises the neurohaemal contact zone or urophysis. The neurosecretory terminals were engorged with secretory granules in these fishes from a freshwater environment. The non-neuronal component of the urophysis also was examined in this study. These glial cells were dispersed throughout the urophysis. Most often they were found ensheathing the terminal neurosecretory processes. These cells showed a resemblance to neurohypophysial pituicytes. Although mentioned in the literature, these glial cells have not been examined with the electron microscope. The ultrastructural features of the glial cells and their relation to the neurosecretory constituents are described in this report.     

N- and R-cadherins expression in the rat sciatic nerve demonstrated by postembedding immunogold method on semi-thin sections

The calcium-dependent adhesion molecules N- and R-cadherins take part in the morphogenesis of the nervous system. The data about their localization in the peripheral nerves postnatum are limited and somewhat controversial. In the present study, a new methodological approach to the immunohistochemical cadherin detection has been applied. The sciatic nerves of 10-day-old rats were immunotested on semi-thin Epon sections by anti-N- and R-cadherins polyclonal antibody and visualized by IgG conjugated to colloidal gold and silver amplification. In the myelinated nerve fibers, an immuno-like activity was associated with the axoplasm and axolemma, the surface of the myelinating Schwann cells and their extracellular matrix. The units of nonmyelinated fibres and their associated glial cells were also intensely immunostained. The myelin sheaths and the glial cytoplasm were immunonegative. The significance of the localization is discussed in regard to the possible role of N- and R-cadherins in the anatomical and functional compartmentation of peripheral nerves.     

The fine structure of choroid plexus carcinoma

The ultrastructural findings of two cases of choroid plexus carcinoma occurring in infants 11 and 24 months old are discussed. Both had a similar ultrastructural pattern which demonstrated a clear polygonal cytoplasm, cell polarity towards the lumina, junctional complexes and evidence of a basement membrane, all of which point to an epithelial origin for the proliferating cells. Abundant immature cellular characters such as polyribosomes, glycogen granules and hypertrophied rough endoplasmic reticulum were found. The importance of the differential dignosis from ependymoblastoma and benign choroid plexus papilloma is emphasized. In the former, the major differentiating features are the existence of abundant glial neuropil and cellular characters of glial differentiation, whilst in the latter, cytoplasmic features demonstrate cellular maturity.     

Cell types in the pineal gland of the horse: an ultrastructural and immunocytochemical study

A combined ultrastructural and immunocytochemical study was performed on the pineal gland of the horse in order to identify the cell types present and describe their characteristics. Comparisons have been made with other mammals. Two main cell types are present: pinealocytes and glial cells. Pinealocytes display different degrees of electron density in the nucleus and the cytoplasm, yet no ultrastructural feature supports the idea of separate populations. Putative secretory materials are stored in vesicles related to the Golgi apparatus. A variety of electron-dense bodies are present in the cytoplasm. Interstitial cells responding to anti-GFAP (glial fibrillary acid protein) and anti-vimentin antibodies, but not to anti-neuronal 200-kD protein antibodies, are located close to the perivascular spaces and connective septa. Morphological and immunocytochemical features support classifying them as astrocytes, probably protoplasmic. The presence of a cavity lined with pericytes, putatively a remnant of the embryonic lumen of the organ, is a consistent finding and may relate to the third ventricle.     

Ultrastructure of the nervous system.

Within the nervous system, five cell types were studied by the electron microscope: the neuron, astrocyte, oligodendroglia, microglia and ependymal cells. The neurons are characterized by the Nissl substance within the perikaryon, the neurotubules and the neurofilaments. The three types of glial cells can be distinguished from one another. The astrocyte has relatively lucent cytoplasm and contains filaments which extend into long cytoplasmic processes. The oligodendroglia has electron-dense cytoplasm with thin extensions wedged between the cellular processes of the neuropil. The microglia cytoplasm is similar to the oligodendroglia but can be differentiated by the peculiar convoluted outline of the nuclear membrane and the clear space which separate the nucleus from the cytoplasm. The ependymal cell has characteristic microvilli and cilia at its ventricular surface. The cytoplasm contain parabasal bodies and tight junctions between adjacent cells. In the peripheral nerve, the axon and the myelin sheath are located within the Schwann cells. The terminal nerve endings from enlargements filled with synaptic vesicles. Electron microscopy of the nervous system is not only essential for research, but is also helpful in diagnosis of certain diseases.     

Retinal pigment epithelium melanin granules are phagocytozed by Müller glial cells in experimental retinal detachment

The ability of retinal Müller glial cells to perform phagocytosis in vivo is studied in a rabbit model of experimental retinal detachment where pigment epithelial cells are occasionally detached together with the neural retina. While macrophages and/or microglial cells phagocytoze most of the cellular debris at the sclerad surface of the detached retinae, some Müller cells accumulate melanin granules. The granules are virtually intact at the ultrastructural level, and are surrounded by a membrane. They are often located close to the sclerad end of the cells, but some are distributed throughout the outer stem process up to the soma. It is concluded that rabbit Müller cells in vivo are capable of phagocytosis and of transporting the phagocytozed material within their cytoplasm.     

Abnormalities of smooth muscle, basal laminae, and nerves in the aganglionic segments of the bowel of lethal spotted mutant mice

The terminal portion of the bowel of the lethal spotted mutant mouse (ls/ls) lacks an enteric nervous system due to the failure of neural crest precursors to colonize this region during embryonic life. As a result, the mouse develops congenital megacolon. We have postulated that the defect occurs because the microenvironment of the aganglionic segment is segmentally abnormal and does not permit the migration and/or survival of the enteric neural or glial precursors in the affected zone. We have examined the terminal segment of adult ls/ls and control mice by light and electron microscopy to determine if the defect is associated with identifiable structural abnormalities that persist to maturity. A striking abnormality is an overgrowth of the muscularis mucosa in the adult ls/ls mouse, particularly in the outer longitudinal layer. Electron microscopy also reveals an extensive thickening of the basal lamina around smooth muscle cells. In addition, nerves that are derived from fibers that are extrinsic to this area are abnormal. Large bundles of nerve fibers, some of which contain myelinated axons, large-caliber unmyelinated axons, and abundant collagen, are prominent in the intermuscular region of the aganglionic segments and often reach into the submucosa. The supporting cells of the unmyelinated and myelinated nerves in the aganglionic segment have voluminous perineural cytoplasm typical of immature Schwann cells. They also exhibit intermediate filaments in their cytoplasm. Otherwise they have the typical morphology of peripheral Schwann cells, rather than enteric glia, including individual ensheathment of axons and a surrounding basal lamina. We suggest that the extracellular matrix and/or cells of mesenchymal origin of the terminal bowel of the ls/ls mouse may prevent the ingrowth of the normal precursors of the glia as well as neurons of the enteric nervous system, but may permit or even encourage the ingrowth of abnormal numbers of extrinsic axons.     

不同禁水时间老龄大鼠下丘脑大细胞分泌神经元的免疫电镜观察

电镜观察了不同禁水时间老龄大鼠下丘脑视上核和室旁核分泌神经元，即加压素和催产素神经元及胶质细胞的超微结构变化，结果显示，禁水6与12h后，上述两核团中的分泌神经元胞体增大，胞质中粗面内质网（RER）排列紧密且规则，高尔基（Golgi)器的未成熟分泌颗粒及神经分泌颗粒增多，轴突内神经分泌颗粒少见，胶质细胞成份减少，突起回缩；相邻两神经元胞膜直接接触，质膜并列现象及突触增多，而在禁水24h后，神经元胞体内的神经分泌颗粒有减少，轴突中的神经分泌颗粒却增多并聚集成膨大区域，以上结果提示老龄大鼠视上核和室旁核的分泌神经元在禁水时其合成激素的功能是活跃的，而且催产素神经元的结构变化与加压素神经元结构变化是相似的，胶质细胞的结构变化为神经元质膜并列及突触的形式提供了有利条件。     

Ventrally emigrating neural tube (VENT) cells: a second neural tube-derived cell population

Two embryological fates for cells of the neural tube are well established. Cells from the dorsal part of the developing neural tube emigrate and become neural crest cells, which in turn contribute to the development of the peripheral nervous system and a variety of non-neural structures. Other neural tube cells form the neurons and glial cells of the central nervous system (CNS). This has led to the neural crest being treated as the sole neural tube-derived emigrating cell population, with the remaining neural tube cells assumed to be restricted to forming the CNS. However, this restriction has not been tested fully. Our investigations of chick, quail and duck embryos utilizing a variety of different labelling techniques (DiI, LacZ, GFP and quail chimera) demonstrate the existence of a second neural tube-derived emigrating cell population. These cells originate from the ventral part of the cranial neural tube, emigrate at the exit/entry site of the cranial nerves, migrate in association with the nerves and populate their target tissues. On the basis of its site of origin and route of migration we have named this cell population the ventrally emigrating neural tube (VENT) cells. VENT cells also differ from neural crest cells in that they emigrate considerably after the emigration of neural crest cells, and lack expression of the neural crest cell antigen HNK-1. VENT cells are multipotent, differentiating into cell types belonging to all four basic tissues in the body: the nerve, muscle, connective and epithelium. Thus, the neural tube provides at least two cell populations--neural crest and VENT cells--that contribute to the development of the peripheral nervous system and various non-neural structures. This review describes the origin of the idea of VENT cells, and discusses evidence for their existence and subsequent fates.     

Caspase-3在成年大鼠中枢神经系统分布的免疫组织化学研究

采用免疫组织化学ABC法观察caspase3在成年大鼠中枢神经系统不同区域内的分布。caspase3阳性反应产物主要分布于脊髓前角和侧角大型运动神经元及中型神经元的胞浆、胞核及突起;脊髓后角及灰质连合的中、小型神经元的胞核及胞浆亦可见caspase3阳性反应产物;脊髓白质内的星形胶质细胞、小胶质细胞及少突胶质细胞的胞核和胞浆,caspase3阳性反应产物呈强阳性反应。在延髓内,caspase3阳性反应产物定位于中央灰质、三叉神经尾侧亚核和中央网状核内中型神经元的胞浆。大脑皮层各区内的caspase3阳性反应产物集中分布于ⅢⅤ层锥体细胞的胞浆,呈弱阳性反应。海马的CA1、CA2、CA3、CA4区的锥体细胞层的细胞,胞浆亦呈弱阳性反应。小脑内,以Purkinje细胞胞浆着色为主。乳头体核、尾状核、豆状核、嗅前核后部、嗅结节及丘脑等部位亦可见caspase3阳性神经元。本研究的结果说明caspase3在中枢神经系统内广泛分布,且在不同部位神经元的亚细胞分布存在差异。