A review of FMRFamide- and RFamide-like peptides in metazoa

Neuropeptides are a diverse class of signalling molecules that are widely employed as neurotransmitters and neuromodulators in animals, both invertebrate and vertebrate. However, despite their fundamental importance to animal physiology and behaviour, they are much less well understood than the small molecule neurotransmitters. The neuropeptides are classified into families according to similarities in their peptide sequence; and on this basis, the FMRFamide and RFamide-like peptides, first discovered in molluscs, are an example of a family that is conserved throughout the animal phyla. In this review, the literature on these neuropeptides has been consolidated with a particular emphasis on allowing a comparison between data sets in phyla as diverse as coelenterates and mammals. The intention is that this focus on the structure and functional aspects of FMRFamide and RFamide-like neuropeptides will inform understanding of conserved principles and distinct properties of signalling across the animal phyla.     

The relationship between complex vesicles, dense-cored vesicles and dense projections in cortical synaptosomes

Cortical synaptosomes fixed in unbuffered OsO4 and glutaraldehyde have been block-stained with phosphotungstic acid (PTA) in order to investigate the relationship between complex vesicles and dense projections. It is concluded that the shell of the complex vesicles contributes to the formation of dense projections and that, in addition, there is a correspondence between this shell and the previously described presynaptic network. The process by which complex vesicles become associated with dense projections appears to be accentuated by electrical stimulation of the synaptosomes.     

Ultrastructural localization of monoamines and peptides in rat area postrema

An overall schema for the synaptic interactions of monoaminergic and peptidergic neurons and their relation to the ventricle and to blood vessels within the rat area postrema is presented. The specific markers include: 1) the immunocytochemical localization of the catecholamine-synthesizing enzyme tyrosine hydroxylase, and the neuropeptides enkephalin and substance P; and 2) the radioautographic localization of [3H]serotonin (5-hydroxytryptamine) and 3H-labeled amino acids anterogradely transported from the nodose ganglion.     

Ultrastructural Studies of the Development of Nerves in Hydra

Three types of mature epidermal neurons and several of theirdifferentiating stages aie presented in this ultrastructuralstudy. Each of the three types, neurosensory, neurosecretory,and ganglionic cells, is derived from interstitial cells, (i)Mature neurosensory cells contain elongated nuclei, a well-developedcilium in each cell, and membrane-bounded neurosecretory droplets(700–1300 A in diameter). There may be two or more neuritesin which are numerous microtubules, glycogen particles, ribosomesand many neurosecretory droplets, (ii) Mature neurosecretorycells closely resemble neurosensory cells, except that no ciliumis present. The perikarya contain small, dense nuclei, neurosecretorydroplets (850–1300 A in diameter), mitochondria, glycogenparticles, and microtubules. Active Golgi complexes are presentin both cell types. The nemites are similar to those describedfor neurosensory cells, (iii) Mature ganglionic cells are bipolaror multipolar. The small, dense nuclei are surrounded by a smallamount of cytoplasm. The neurites contain mostly microtubules;a few mitochondria, ribosomes, and glycogen particles are alsopresent, but there are no secretory droplets. To date, only neurosensory and neurosecretory cells have beenobserved in the gastiodermis. They are structurally indistinguishablefrom their epideimal counterparts. A significant finding is that three types of synapses—neuromuscular,neuronematocyte, and interneuronal—are identified in boththe epidermal and gastrodermal neurons.     

Ultrastructural localization of a new neuronal peptide (VIP).

The vasoactive intestinal polypeptide (VIP) represents a neuronal peptide of wide-spread occurrence. By electron microscopic immunocytochemistry VIP has been localized to the granules of the terminals of p-type neurons. This localizations lends support to the postulate that VIP may be a neurotransmitter.     

Ultrastructural localization of egg-laying prohormone-related peptides in the atrial gland of Aplysia californica

The atrial gland is an exocrine organ that secretes into the oviduct of Aplysia californica and expresses three homologous genes belonging to the egglaying hormone gene family. Although post-translational processing of the egg-laying hormone precursor in the neuroendocrine bag cells has been examined in detail, relatively little is known about the post-translational processing of egg-laying hormone-related gene products in the atrial gland. A combination of morphologic techniques that included light-microscopic histology and immunocytochemistry, transmission electron microscopy, and immuno-electron microscopy were used to localize egg-laying hormone-related peptides in the atrial gland and to evaluate the characteristic morphology of their secretory cells. Results of these studies showed that there were at least three major types of secretory cells in the atrial gland (types 1–3). Significantly, of these three cell types, only type 1 was immunoreactive to antisera against egg-laying hormone-related precursor peptides. The immunoreactivity studies established that all three egg-laying hormone-related precursor genes are expressed in type-1 cells and indicated that the processing of these precursors also occurs within the secretory granules of this cell type. Evidence was also obtained that proteolytic processing of the egg-laying hormone-related precursors differed significantly from that observed in the bag cells. In contrast to the bag cells, the NH₂-terminal and COOH-terminal products of the egg-laying hormone-related precursors of the atrial gland were not sorted into different types of vesicles.     

Ultrastructural localization of a new neuronal peptide (VIP)

Summary The vasoactive intestinal polypeptide (VIP) represents a neuronal peptide of wide-spread occurrence. By electron microscopic immunocytochemistry VIP has been localized to the granules of the terminals of p-type neurons. This localization lends support to the postulate that VIP may be a neurotransmitter.A brief report of the present work was given at the First International Symposium on Gastrointestinal Hormones, Asilomar, California, October 1976     

Fusion failure of dense-cored proacrosomal vesicles in an inducible mouse model of male infertility

The acrosome is a specialized secretory vesicle located in the head of spermatozoa and has an essential role during fertilization. This organelle and the sperm nucleus have aberrant morphologies in forms of male infertility in humans (teratozoospermia), often associated with poor motility (asthenoteratozoospermia). To further our understanding of the aetiology of these conditions, we have performed a pathological investigation of a model of asthenoteratozoospermia that can be induced in mice by N-butyldeoxynojirimycin (NB-DNJ). We have found that, in mice treated with NB-DNJ, instead of an acrosome forming over the round spermatid nucleus, multivesicular bodies (MVB) accumulate in the vicinity of this nucleus. Electron microscopy has revealed that proacrosomic vesicles or granules (PAG) secreted during the Golgi phase of spermiogenesis do not fuse together to form an acrosomic vesicle, but rather attach transiently to the spermatid nucleus. Immunocytochemistry has shown that acrosomal membrane proteins and cytosolic acrosome-associated proteins are redirected to MVB in affected testes, whereas glycoproteins originating in the dense core of the PAG are degraded. Thus, the major effect of NB-DNJ is to inhibit membrane fusion of Golgi-derived secretory vesicles destined for acrosome formation, raising the possibility that these vesicles are critically affected in forms of (astheno)teratozoospermia.     

Effects of blinding on the ultrastructure of mouse pinealocytes with particular emphasis on the dense-cored vesicles

Summary The mammalian pineal is thought to produce an antigonadotropic principle under conditions of reduced photoperiod, constant darkness or blinding by optic enucleation. A number of previous studies on mammalian pineals have suggested that the dense-cored vesicles present in pinealocytes may represent morphological evidence of secretory activity.In the present study the ultrastructure of pinealocytes was studied in adult Charles River CD-1 mice blinded by optical enucleation. By one month following optic enucleation the mean number of dense-cored vesicles in the cytoplasm of pinealocytes adjacent to pericapillary spaces had significantly decreased by 55% when compared with intact controls, and remained at this low level at two months and six months. A relative increase in the proportion of large agranular vesicles and an increased number of large, irregular vacuoles was observed also in the pinealocytic polar processes of blinded mice. When compared to control mice the pinealocytic Golgi regions appeared to be hypertrophied in blinded mice. The apparent stimulation of pinealocytic organelles coupled with the observed decrease in dense-cored vesicles suggest an increased synthesis and release of secretory product.Supported in part by NIH Grant No. HD 08759     

Ultrastructural evidence for the presence of nerve cells in the gastrodermis of Hydra

Summary Two types of nerve cells, namely, neurosensory and neurosecretory cells have been identified and described in the gastrodermis of Hydra pseudoligactis. The morphological criteria used for the identification of gastrodermal nerves are based on those presented previously for epidermal nerves. The third type of nerve cell in the epidermis, ganglionic cells, was not observed in these studies. The distribution, function and origin of gastrodermal nerve cells are discussed briefly.With the technical assistance of Linda M. Bookman.     

Bilayer localization of membrane-active peptides studied in biomimetic vesicles by visible and fluorescence spectroscopies.

Depth of bilayer penetration and effects on lipid mobility conferred by the membrane-active peptides magainin, melittin, and a hydrophobic helical sequence KKA(LA)7KK (denoted KAL), were investigated by colorimetric and time-resolved fluorescence techniques in biomimetic phospholipid/poly(diacetylene) vesicles. The experiments demonstrated that the extent of bilayer permeation and peptide localization within the membrane was dependent upon the bilayer composition, and that distinct dynamic modifications were induced by each peptide within the head-group environment of the phospholipids. Solvent relaxation, fluorescence correlation spectroscopy and fluorescence quenching analyses, employing probes at different locations within the bilayer, showed that magainin and melittin inserted close to the glycerol residues in bilayers incorporating negatively charged phospholipids, but predominant association at the lipid-water interface occurred in bilayers containing zwitterionic phospholipids. The fluorescence and colorimetric analyses also exposed the different permeation properties and distinct dynamic influence of the peptides: magainin exhibited the most pronounced interfacial attachment onto the vesicles, melittin penetrated more into the bilayers, while the KAL peptide inserted deepest into the hydrophobic core of the lipid assemblies. The solvent relaxation results suggest that decreasing the lipid fluidity might be an important initial factor contributing to the membrane activity of antimicrobial peptides.     

Ultrastructural studies of some chitinozoan vesicles

Four species of Ordovician Chitinozoa from Öland, Sweden, Cyathochitina stentor, Desmochitina minor, Lagenochitina esthonica , and Lagenochitina tumida , have been investigated by transmission electron microscopy. There is very great variation in the vesicle wall ultrastructure of the specimens studied. The texture of the vesicle wall can be homogeneous or granular and can also contain denser zones. Ultrastructures such as pore canals and 'empty spaces' have been revealed. The systematic affinity of the Chitinozoa is discussed, and it is concluded that chitinozoans display typical metazoan structures represented in eggs and egg capsules of marine invertebrates and possibly also in fish.     

Ultrastructural localization of acetylcholinesterase

Summary A method is described allowing localization of acetylcholinesterase (AChE) by both light and electron microscopy. During the reaction lead thio-diacetyl is decomposed, and therefore precipitated as PbS in the presence of native-SH group produced by the hydrolysis of acetylthiocholine perchlorate. The reaction takes place at neutral pH, since improves the sensitivity of AChE localizations. Application of the method to parasympathetic neurons showed that AChE was mainly localized in the rough endoplasmic reticulum of the perikaryons. No reaction was visible in glial cells. AChE was also localized on the plasma membrane of parasympathetic neurons. In mouse embryo muscles AChE activity was seen to be high and was not yet restricted to the synaptic area. The well developed Schwann cells accompanying the neurites displayed constant AChE activity on their plasma membrane.Supported by a grant of INSERM C.R.L. N⁰ 79-5-318-6     

Fine structure of dense-cored vesicles in glomus cells of rat carotid body after fixation with permanganate or glutaraldehyde

Summary Glomus (Type I) cells of the carotid body of adult rats were studied electron microscopically after fixation with potassium permanganate or with glutaraldehyde and osmium tetroxide. Two permanganate fixation methods (using Krebs-Ringer-glucose, pH 7.0, or acetate buffer, pH 5.0) were compared. Numerous dense-cored vesicles were observed only in about one tenth of the glomus cells when neutral permanganate was used for fixation, although all glomus cells showed such vesicles after fixation with glutaraldehyde and osmium tetroxide. Numerous vesicles with a dense core were observed in about one third of the cells after fixation with acid potassium permanganate. With this fixation, small dense-cored vesicles similar to those in adrenergic nerve terminals were occasionally seen in the cytoplasm of glomus cells. It is tentatively concluded that the amine-storing vesicles of the carotid body are different from those in the small intensely fluorescent (SIF) cells and those in adrenergic nerve terminals.     

Ultrastructural heterogeneity of vesicles in the brush border of enterocytes

Ultrastructural properties of microvesicles, situated in the brush border of rat's enterocytes were studied in normal condition, upon toxic action of cholic acid and of CCl4. Heterogeneity of the objects under study was demonstrated, properties of their excretion by enterocytes are emphasized. Particular criteria of differentiation of excretory vesicles in the parietal zone of epithelium of rat's ileum were given.     

Ultrastructural localization of calcium in matrix vesicles and preodontoblasts of developing rat molar tooth germs during initial dentinogenesis

We investigated the ultrastructural localization of calcium in progenitor predentine and preodontoblasts of developing rat molar tooth germs using the potassium pyroantimonate technique. At the precalcification stage, antimonate reaction product was sparsely, randomly distributed in the preodontoblasts and in the progenitor predentine but no significant reaction could be noticed associated with matrix vesicles. At the matrix vesicle calcification stage, large amounts of antimonate reaction product tended to be localized in the region adjacent to the distal, outer surface membrane of preodontoblasts in which moderate antimonate reaction activity could be observed in mitochondria. Strong antimonate reaction was detected preferentially on the outer surface membrane of some matrix vesicles at this stage. At the subsequent collagen calcification stage, definite antimonate reaction was no longer seen within mitochondria of the late preodontoblasts, instead precipitate was mainly distributed in Golgi area, secretory granules and lateral intercellular spaces. It is suggested that although matrix vesicles contain few calcium capable of reacting to antimonate immediately after their biogenesis, subsequently, large amounts of calcium are accumulated associated with the outer surface membrane of matrix vesicles in the extracellular matrix.