Ultrastructural studies on the secretory cycle of the neurosecretory cells and the formation of herring bodies in the paraventricular nucleus of the rat

Summary The ultrastructure of the neurosecretory cells in the paraventricular nucleus of the normal male rat was studied by electron microscopy during various functional states. Four morphologically distinct types of neurosecretory cells were observed. It appears that they do not represent different classes of cells but different phases of secretory activity of a single cell type. The perikarya of the neurosecretory cells show a definite cycle of formation and transportation of secretory granules. We have designated the phases of this cycle as: (1) phase of synthesis, (2) phase of granule production, (3) phase of granule storage and (4) phase of granule transport. The neurosecretory granules appear to be moved in bulk into the axons, forming a large axonal swelling filled with granules as a result of one cycle in the neurosecretory process. Thus it may be postulated that a secretory cycle in the perikaryon of the neurosecretory cell seems to result in the formation of a Herring body in its axon, and that its content is then conveyed to the posterior pituitary.     

Cytological aspects of different nerve cell somata in the buccal ganglia of Helix pomatia L.

In early September most of the neurons of the buccal ganglia of Helix pomatia contain neurosecretory material as membrane bound granules. There is only one, in exceptional cases 2 types of granules per cell. This suggests that different types of granules do not change into one another, and that each granule type contains a different secretory product. One granule type contains PAF-positive neurosecretory material, another one catecholamines, but most of the granules cannot be associated with special substances. The identified giant neurons B1-B4 contain granules in less density than the smaller neurons. B1 and B2 resemble each other in their granule type, whereas both B3 and B4 differ from B1 and B2.     

Purification of bovine neurosecretory granule membranes by density gradient centrifugation

A procedure for the subfractionation of neurosecretory granules into membrane and content components is described. The procedure involves the hypotonic lysis of the secretory granule fraction and further purification of the membranes by centrifugation through a discontinuous sucrose gradient. The neurosecretory granule membranes represented 5.2% of the total proteins of the neurosecretory granule fraction and were highly enriched in cytochrome b561. Electron microscopic analysis of the purified membranes showed vesicles devoid of electrodense content.     

Are mechanisms of exocytosis neurotransmitter specific?

In their review, Langley and Grant (1997) investigate the question whether mechanisms of exocytosis are neurotransmitter specific. There is now much evidence that the mechanisms governing the exocytosis of the two principal storage organelles—granules (large dense core vesicles) and electron-lucent vesicles—differ. But much less is known concerning potential differences in the release mechanisms of electron-lucent vesicles that store different types of fast neurotransmitters or of granules in different types of neurons. It is an open question whether there is a unifying control mechanism for the exocytosis of, for example, a peptide-containing granule of a glutamatergic neuron, a chromaffin granule, a noradrenergic granule or a granule from a neurosecretory neuron in the pituitary. The small electron-lucent synaptic vesicles of various kind apparently share common molecular components of regulated release. They carry the calcium sensor synaptotagmin, small GTP-binding proteins of the rab3 group or the v-SNARE synaptobrevin. Nevertheless, there may be differences in the regulatory mechanisms. This concerns the type of calcium channel involved or the absence of some of the presynaptic molecules such as rab3a, synapsin I or the t-SNAREs SNAP-25 or syntaxin from distinct types of neurons or sensory cells.     

鲫鱼尾部神经分泌系统显微和亚显微结构的季节性变化

鲫鱼尾部神经分泌系统的神经分泌细胞和它的轴突中可观察到各种不同电子密度的颗粒。在性腺各个不同的发育阶段,该系统的分泌物具有累积、充满、释放和恢复这样一种周期性变化,由此说明鲫鱼的尾部神经分泌系统和它的生殖有关。     

Morphofunctional basis of neurosecretory cell plasticity

Mass accumulation and storage of neurosecretory products are typical only for nonapeptidergic elements, as it has been shown by our study of the structure and function in neurosecretory cells of different nature. All liberinergic, statinergic and monoaminergic neurosecretory cells keep constancy in the state of high functional activity of extrusive processes at normal conditions. Morpho-functional features of these elements principally differ from those of nonapeptidergic neurosecretory cells, which are characterized by remarkable secretory cycles. The extremely large size of elementary secretory granules, maximum development of the Herring bodies, various modes of secretion, secretory and extrusive cycles in neurosecretory function, and massive accumulation of neurosecretory granules occurring in neurosecretory terminals finally, all these characters are considered to be the primary features of a high plasticity of the nonapeptidergic neurosecretory cell. A high reactivity of nonapeptidergic neurosecretory cells has been demonstrated here by the quantitative ultrastructural research of the dynamics of functional activity of neurosecretory terminals at both experimental and physiological stressful states. The highest plasticity of nonapeptidergic neurosecretory cells compared to all other neurosecretory cell types may be provided by their ability to restore the initial law level of functional activity, referred to as "functional reversion".     

Regulation of secretory granule size by the precise generation and fusion of unit granules

Morphometric evidence derived from studies of mast cells, pancreatic acinar cells and other cell types supports a model in which the post-Golgi processes that generate mature secretory granules can be resolved into three steps: (1) fusion of small, Golgi-derived progranules to produce immature secretory granules which have a highly constrained volume; (2) transformation of such immature granules into mature secretory granules, a process often associated with a reduction in the maturing granule’s volume, as well as changes in the appearance of its content and (3) fusion of secretory granules of the smallest size, termed ‘unit granules’, forming granules whose volumes are multiples of the unit granule’s volume. Mutations which perturb this process can cause significant pathology. For example, Chediak–Higashi syndrome / lysosomal trafficking regulator (CHS)/(Lyst) mutations result in giant secretory granules in a number of cell types in human beings with the Chediak–Higashi syndrome and in ‘beige’ (Lyst^bg/Lyst^bg) mice. Analysis of the secretory granules of mast cells and pancreatic acinar cells in Lyst-deficient beige mice suggests that beige mouse secretory granules retain the ability to fuse randomly with other secretory granules no matter what the size of the fusion partners. By contrast, in normal mice, the pattern of granule–granule fusion occurs exclusively by the addition of unit granules, either to each other or to larger granules. The normal pattern of fusion is termed unit addition and the fusion evident in cells with CHS/Lyst mutations is called random addition. The proposed model of secretory granule formation has several implications. For example, in neurosecretory cells, the secretion of small amounts of cargo in granules constrained to a very narrow size increases the precision of the information conveyed by secretion. By contrast, in pancreatic acinar cells and mast cells, large granules composed of multiple unit granules permit the cells to store large amounts of material without requiring the amount of membrane necessary to package the same amount of cargo into small granules. In addition, the formation of mature secretory granules that are multimers of unit granules provides a mechanism for mixing in large granules the contents of unit granules which differ in their content of cargo.     

Fine structure of the neurohemal sinus gland of the shore crab,Carcinus maenas,and immuno-electron-microscopic identification of neurosecretory endings according to their neuropeptide contents

Summary The sinus gland of the shore crab, Carcinus maenas, is a compact assembly of interdigitating neurosecretory axon endings abutting upon the thin basal lamina of a central hemolymph lacuna. Four types of axon endings are distinguishable by the size distribution, shape, electron density and core structure of their neurosecretory granules. One additional type of axon ending is characterized by electron-lucent vacuoles and vesicles. The axon profiles are surrounded by astrocyte-like glial cells. Various fixations followed by epoxy- or Lowicrylembedding were compared in order to optimize the preservation of the fine structure of the granule types and the antigenicity of their peptide hormone contents. By use of specific rabbit antisera, the crustacean hyperglycemic, molt-inhibiting, pigment-dispersing, and red-pigment-concentrating hormones were assigned to the four distinct granule types which showed no overlap of immunostaining. Epi-polarization microscopy and ultrathin section analysis of immunogold-stained Lowicrylembedded specimens revealed that immunoreactivity to Leu-enkephalin and proctolin is co-localized with moltinhibiting hormone immunoreactivity in the same type of granule. The size and core structure of the immunocytochemically identified granule types vary little with the different pretreatments but, in some cases, to a statistically significant extent. The present results are compared with those from earlier studies of sinus glands in different crustaceans. The methods of granule identification used in this study supplement the classical approach in granule typing; they are easier to perform and more reliable for the analysis of release phenomena in identified secretory neurons supplying the neurohemal sinus gland.     

Cytochemical ultrastructural study of the glycoproteins in the rat hypothalamo-post-pituitary complex]

The results obtained with various methods applied to the cytochemical detection of carbohydrates at an ultrastructural level, confirm the existence of glycoproteins in neurosecretory material in the neurohypophysis as well as in the hypothalamic magnocellular nuclei. This glycoproteic component, however, is not present in all the secretory granules and, according to their cytochemical behaviour, it is possible to distinguish two types of neurosecretory fibres: one where all the granules respond negatively; the other where most of the granules are reactive. The existence of two types of neurons corresponding to these two fibres cannot yet be asserted, but seems very likely, perhaps connected with the hormonal duality of the magnocellular nuclei. The reactions are also positive on the Golgi apparatus, in accordance with its function in glycoprotein synthesis. But the difference of reactivity between the Golgi cisternae and the neurosecretory product suggests that glycoprotein synthesis is still going on in the neurosecretory granules outside the Golgi area.     

Licht- und elektronenmikroskopische Untersuchung von neurosekretorischen Zellen im Gehirn der Eintagsfliege Ephemera danica Müll. (Ephemeroptera: Ephemeridae)

In the brains of the males the amount of stained secretory material was nearly constant during the last four instars. In the females a decrease in this neurosecretory product in the last nymphal and subimaginal stage could be observed, followed by an increase in the imagines. In the final nymphal stage four types of neurosecretory cells (nsc) were found in the medial protocerebral cell group, showing differences in shape, size, and the contents of the cytoplasm, especially of the secretory granules. In addition to the medial nsc, some cells in the frontal part of the brain and in the deutocerebrum are described. They contain electron-opaque granules and probably have a neurosecretory function. The secretory product of the medial nsc is transported along the axons of the nervus corporis cardiaci 1 (ncc1), an unpaired nerve tract on the ventral side of the brain. Leaving the brain the ncc1 immediately enters the corpus cardiacum. Connections between secretory granules and neurotubuli point to an important role for the neurotubuli in the transport of secretory material.     

Ultrastructure of the protocerebral neurosecretory cells of larval Galleria mellonella, in situ and after culture of the brain in vitro

Three major groups of neurosecretory cells are described in the larval brain of Galleria mellonella at two different times during the last larval instar and in larval brains after 72 hr of culture in vitro. The medial group in vivo consists of four distinct neurosecretory cell types, based on characteristic size and morphology, while the posterior and lateral groups each contain a single distinct type of neurosecretory cell. Morphological differences between the same neurosecretory cells at the different times during the last instar are most apparent in the lateral L-1 cells and in the medial M-2 cells, where pleiomorphism is particularly evident in the size, density and accumulations of neurosecretory granules. The only neurosecretory cells in which apparent synthesis of neurosecretory granules is still observed after culture of the brain in vitro are the medial M-2 cells. The other neurosecretory cell types show no accumulation of neurosecretory granules nor new synthesis of neurosecretory material, but are similar to neurosecretory cells in the brain in vivo in all other respects. The morphology of the neurosecretory cells in the larval brain in vivo and in vitro is discussed in relation to their appearance at the light microscopic level and to a known neurohormonal function of the brain which is maintained during 72 hr in vitro.     

Ultrastructural Analysis of the Neuroendocrine Apparatus of Oncopeltus fasciatus (Heteroptera)

• 1 In Oncopeltus fasciatus, the A-cells of the pars intercerebralis and their tracts are stainable in situ with the performic acid-victoria blue (PAVB) method. The axons from these cells, after traversing the corpus cardiacum, terminate in the anterior part of the aorta which thus serves as the neurohemal organ.
• 2 Ultrastructurally, four types of secretory neurons are distinguishable in the pars intercerebralis region: pic-I with granules measuring 1000–3000 Å in diameter; pic-II with granules of irregular size and shape, the elongate ones showing mean dimensions of 2400 × 1400 Å; pic-III with less electron-dense granules measuring 1000–2700 Å in diameter; pic-IV, present not only in the pars intercerebralis but also in adjacent regions of the brain, with variable proportions of granules measuring 700–1800 A and dense-cored vesicles measuring 1000–2400 Å.
• 3 The nervi corporis cardiaci contain at least three types of neurosecretory axons, based on granule content, presumably representing pic-I, pic-II and pic-III neurons.
• 4 The wall of the aorta contains endings of at least three distinct types, again representing pic-I, pic-II and pic-III neurons, and thus provides the neurohemal site for these three types of protocerebral neurosecretory cells. Axons of pic-IV neurons appear to enter the cerebral neuropil.
• 5 The corpus cardiacum is composed of two types of parenchymal secretory cells, with electron-dense granules measuring 1300–3000 Å and 1000–2300 Å in diameter, respectively. The corpus cardiacum also contains interstitial cells and some axons of extrinsic origin, with and without granules.
• 6 The corpus allatum may be paired or median, and receives a small number of at least two types of axons. The corpora allata of some reproducing females show a large number of PAVB-stainable inclusions which appear to be modified cytoplasmic organelles, but are definitely not neurosecretory material.
• 7 The hypocerebral ganglion is composed of two types of secretory-appearing neurons and glial cells. The two neuronal types contain secretory granules, 1000–3000 Å and 900–2100 Å in diameter, respectively. Axons of the recurrent nerve also may contain occasional granules.
• 8 In this heteropteran insect, the two principal functions of the corpus cardiacum appear to be spatially separated: the neurohemal function is subserved by the aortic wall, which permits release of material into both the aortic lumen and the hemocoel, and the intrinsic endocrine function is possessed by the parenchymal cells.

     

锯缘青蟹窦腺显微和超微结构研究

借助光学和电子显微镜观察据缘青蟹（Ｓｃｙｌｌａ ｓｅｒｒａｔａ）窦腺的形态结构。窦腺位于眼柄视神经节内髓背侧近外髓处。窦腺呈羹状;腺体壁山神经分泌细胞的末梢和神经胶质细胞组成。神经末梢内充满了电子致密的神经分泌颗粒。根据颗粒的大小、形态及电子致密度等特征,可以区分出４种类型的神经末梢。一些末梢中的多形性颗粒可能是由Ⅱ型末梢中的颗粒转变而成的。一些现象表明,神经分泌物质可以通过胞吐作用或一种类似“顶浆分泌”的方式释放,从而说明神经激素的释放可能是多途径的．     

Ultrastructural study of the pericardial organ-anterior ramifications complex neurosecretory terminals

Summary The pericardial organ-anterior ramifications complexes of Uca pugilator and Callinectus sapidus were studied by transmission electron microscopy. Five morphologically distinguishable groups of granules and two groups of vesicles were identified. These granules and vesicles are present in approximately the same proportions in the pericardial organs and anterior ramifications of both species. Two of the granule groups are never mixed in the same axon terminals and are believed to represent different hormone-protein complexes. The remaining granule and vesicle groups are believed to be products of neurosecretory hormone release. Evidence that at least some of these granules and vesicles arise from intraaxonal release of neurosecretory material is presented.This work was supported by USPHS-NIH Training Grant GM-00669 and by the University of Texas Institutional Grant No. 5 SO 1 RR 05426-11.     

Granulolysis in neurosecretory neurons of the rat supraoptico-posthypophyseal system

Ultrastructural and cytochemical observations on neurosecretory neurons of the rat supraoptico-posthypophyseal systems were made under experimental conditions which resulted in striking changes in the amount of neurosecretory granules and lysosomes. Attention was focused on granulolysis. At the onset of rehydration following a 4 days water deprivation, very active autophagy took place in neurosecretory axons of the neural lobe involving the marked increase in smooth endoplasmic reticulum, microvesicles and neurosecretory granules, although the latter were still very few due to previous depletion. When axonal transport was inhibited by colchicine at the onset of rehydration, granules accumulated in the perikarya while granule reloading of the neural lobe was delayed. However autophagy, although always active in axons, remained scarce in perikarya. Moreover, in the latter there was only slight evidence of crinophagy. Hypophysectomy also induced granule accumulation in the perikarya, although accompanied by little granulolysis. Images indicative of crinophagy as shown by acid phosphatase localization were few and exclusively restricted to perikarya, while autophagy occurred essentially in axons. Autophagy appeared to be the predominant process for granulolysis and might be considered here as an aspect of the general turnover of cell constituents, related to the sudden regression of hyperactivity-induced hyperthrophy, rather than as an expression of a specific regulation of an excess of secretory material.     

Neurosecretory granule formation in ligated axons: additional arguments for a local differentiation from a Golgi apparatus extension

The sorting domain for the different types of granules and small synaptic vesicles in neurosecretion is still largely a matter of debate. Some authors state that an exocytotic process has to precede granule formation. In previous studies, we favoured the idea that neurosecretory packages in terminals are assembled from axonal reticulum membranes simply by differentiation at the axon ending, the axonal reticulum being an extension of the Golgi apparatus. By ligating bovine splenic nerve, a de novo differentiation can be induced. After ligation, granules and granulo-tubular complexes appear. They were immunoreactive for SV2, VMAT2 and synaptobrevin II, which are all known to be highly enriched in large dense granules. Previously the granulo-tubular structures have already been recognized as precursor stadia of neurosecretory granules.It is concluded that at a de novo differentiation, a sorting out and aggregation is taking place of molecules typical for large dense granules. The small dense granules and tubules can be considered unripe, precursor forms of the large dense granules. All this occurs in the absence of signs of exocytosis. The present findings corroborate the view that granule formation occurs via local differentiation at an axon ending.     

Ultrastructure of esophageal glands and their secretory granules in the root-knot nematodeMeloidogyne incognita

Summary The dorsal and subventral esophageal glands and their secretory granules in the root-knot nematodeMeloidogyne incognita changed during parasitism of plants. The subventral esophageal glands shrank and the dorsal gland enlarged with the onset of parasitism. While secretory granules formed by both types of glands were spherical, membrane-bound, and Golgi derived, the granules differed in morphology and size between the two types of glands. Subventral gland extensions in preparasitic second-stage juveniles were packed with secretory granules which varied in diameter from 700–1,100 nm and had a finely granular matrix. Within the matrix of each subventral gland granule was an electron-transparent core that contained minute spherical vesicles. The size and position of the core varied within different granules. Few granules were present in the dorsal gland extension in preparasitic juveniles. The matrix of dorsal gland secretory granules formed during parasitism was homogeneous and more electron-dense than the matrix of subventral gland granules. Subventral gland secretory granules of parasitic juveniles and adult females appeared degenerate.     

ICA 512, an autoantigen of type I diabetes, is an intrinsic membrane protein of neurosecretory granules.

Islet cell autoantigen (ICA) 512 is a novel autoantigen of insulin-dependent diabetes mellitus (IDDM) which is homologous to receptor-type protein tyrosine phosphatases (++PTPases). We show that ICA 512 is an intrinsic membrane protein of secretory granules expressed in insulin-producing pancreatic beta-cells as well as in virtually all other peptide-secreting endocrine cells and neurons containing neurosecretory granules. ICA 512 is cleaved at its luminal domain and, following exposure at the cell surface, recycles to the Golgi complex region and is sorted into newly formed secretory granules. By immunoprecipitation, anti-ICA 512 autoantibodies were detected in 15/17 (88%) newly diagnosed IDDM patients, but not in 10/10 healthy subjects. These results suggest that tyrosine phosphorylation participates in some aspect of secretory granule function common to all neuroendocrine cells and that a subset of autoantibodies in IDDM is directed against an integral membrane protein of insulin-containing granules.     

The neurosecretory system of the eyestalk of Carcinus maenas (Crustacea: Decapoda)

The optic ganglia neurosecretory cells of male and female Carcinus maenas during intermoult are distinguishable into six types based on size, location, appearance and method of secretory material release from the perikaryon. Release occurs via the sinus gland and also, in one case, directly into blood capillaries among the neurosecretory cells themselves. The sinus gland consists of axonal extensions of the neurosecretory cells; no secretory granules are produced there and nuclei observed between the axonal endings are those of ill-defined glial cells.