Histochemistry of neurosecretory cells in the cerebral ganglion of the adult or juvenile sweet water crab, Potamon magnum magnum (Pretzman)]

The histochemistry of the neurosecretory material of the PF-positive neurosecretory cells of the cerebral ganglion of the adult and juvenile crab, Potamon magnum magnum (Pretzman) has been investigated. The PF-positive neurosecretory material is proteinaceous and contains considerable amounts of cystine; it is associated with significant amount of cytoplasmic RNA, lipids and phospholipids and with moderate amounts of carbohydrates, though no glycogen could be detected. The significance of these results is discussed.     

The structure and cytochemistry of the neurosecretory cells in the intertidal gastropod Thais bufo (lamarck)

Three types of neurosecretory cells ('A', 'B' and 'C' cells) have been distinguished in the central ganglion of Thais bufo. A few homogenous groups are met with and the rest are all heterogenous groups. The histochemical observations reveal that the neurosecretory material is rich in carbohydrates, disulphides, protein bound amino groups, glycoprotein and lipids. Thus the neurosecretory material seems to be a lipoprotein--glycoprotein complex.     

SULFHYDRYL AND DISULFIDE GROUPS OF PROTEINS : I. METHODS OF ESTIMATION

1. Methods have been described for reducing protein S-S groups, for oxidizing protein SH groups, and for estimating protein S-S and SH groups. 2. It has been found necessary in estimating the cystine content of proteins by the Folin-Marenzi method to take into account any cysteine that may be present. 3. A method for estimating the cysteine content of proteins has been described. 4. With these methods, estimations have been made of the S-S and SH groups and of the cystine and cysteine contents of a number of proteins. 5. In a denatured, but unhydrolyzed protein, the number of S-S and SH groups is equivalent to the quantity of cystine and cysteine found in the protein after hydrolysis.     

The histochemistry of the clitellum of Tubifex tubifex (annelida: oligochaeta)

The histochemistry of the clitellar glands of Tubiflex tubiflex is reported, together with the ultrastructure of mature secretion units. The distribution and ultrastructure of mucous and granule cells are consistent with published accounts of tubificid citella, thus indicating an absence of intrageneric variation. Mucous cells contain homogenous electron lucent globules which are rich in neutral and acidic mucopolysaccharides and are non-reactive with techniques designed to reveal protein. Type I cells possess structured granules composed of tyrosine-containing basic protein and carboxylated acid mucopolysaccharides. The granules of type II cells are heterogenous and contain several 0.3--0.7 micrometer inclusions of acidic protein with identifiable tyrosine, tryptophan and cystine. The inclusions occur within a matrix composed of neutral mucopolysaccharides and carboxymucin. Type III cell granules possess 0.2--0.5 micrometer inclusions within an electron dense matrix and are rich in neutral and carboxylated acid mucopolysaccharides and basic protein containing tyrosine, tryptophane and cystine. Clitelar carboxymucins are characterized by their stability to beta-glucoronidase, hyaluronidase, neurominidase and acid hydrolysis. A pyridine-extractable lipid moiety within the clitellar secretions was not detected. The results are discussed in relation to cell function during cocoon deposition and, comparatively, with those documented for other oligochaete families.     

The hypothalamo-neurohypophysial system in Indian fresh-water goby, Glossogobius giuris (Ham.)

The hypothalamo-neurohypophysial neurosecretory system in Indian fresh-water goby, Glossogobius giuris (Ham.) has been described. The tractus preoptico-hypophyseus serves the function of a morphological and physiological connection between the hypothalamus and pituitary gland. In addition to main mass of the nucleus preopticus cells (cystine/cysteine bearing), a group of few cells in the hypothalamus has also been observed. These cells are situated posterior to the position of the nucleus preopricus and are CH Ph + ve and AF + ve. The neurosecretory material in the cells of nucleus preopticus is in the form of fine granules. The nucleus lateralis tuberis is absent in the fish under study. The disposition of neurosecretory material is heaviest along the fibres of the neurohypophysis in the region of pars intermedia with which it forms a profuse interdigitation. The fibres usually terminate over the blood vessels. The Herring bodies are noticeable at different levels in the neurohypophysis and pars-distalis. Besides the neurosecretory fibres, Herring bodies, non-stainableneurosecretory fibres and blood vessels, the pituicytes are also present in the neurohypophysis (SAKSENA 1974a, b). The intraaxonal flow of neurosecretory material, the vascularization of the nucleus preopticus and hypothalamo-hypophysial regulatory mechanism have been also discussed.     

Neurosecretory system of the ventral ganglia in the dragonfly,Orthetrum chrysis (selys) (Odonata: Libellulidae)

Summary The neurosecretory cells of the ventral ganglia in the adult dragonfly, Orthetrum chrysis, are classified into A, B, C1 and C2 cells. The neurosecretory material in the ventral ganglia is composed of PAS-positive material with 1-, 2-glycol groups and some proteins. The proteins rich in cystine or cysteine occur predominantly in the A cells, moderately in C cells and negligibly in B cells. Proteins containing arginine occur in A and B cells only, and those containing basic amino acids occur in C2 cells. The neurosecretory pathways and the neurohemal organs are also described.     

Neurosecretory protein synthesis in relation to starvation and feeding in adult male Locusta migratoria

An immunological approach has been used to assess the effects of starvation and subsequent feeding upon the synthesis of neurosecretory protein in adult male Locusta migratoria. Starvation (for 5 days) did not reduce the incorporation of radioactive cystine into neurosecretory protein. However, the rate of neurosecretory transport in starved insects was approximately half that in normal fed insects.Within 1 hr after the start of feeding the rate of neurosecretory protein synthesis more than doubled and the rate at which newly synthesized protein was transported to the CC increased approximately three-fold. The incorporation of cystine into specific (i.e. neurosecretory) protein was always higher, even in starved insects, than into the non-specific proteins (i.e. proteins other than those from the A-cells) of the brain. This difference was maximal at 4 hr after feeding when the incorporation into specific protein was more than 5 times that into non-specific protein.     

Histochemical observations on the giant neurosecretory cells of the thoracic ganglion of the adult and juvenile crabs, Potamon magnum magnum (Pretzman).

The giant neurosecretory cells in the thoracic ganglion of the adult and juvenile crab, Potamon magnum magnum (Pretzman) were histochemically investigated. The secretion is mainly proteinaceous in nature, containing considerable amounts of acid mucosubstances, sulphate esters, lipids and a little carbohydrate but no glycogen. The detailed nature of proteinaceous neurosecretory material in the adult crab was further tested. It appears that the neurosecretory material of these cells contains moderate amounts of sulfhydryl groups and few of disulphide bonds. No trace of tyrosine could be observed. The neurosecretory granules were associated with considerable amounts of cytoplasmic RNA. In general, stronger reactions were obtained in summer and winter than in other seasons.     

A histochemical study of the cells of the ventral cord ganglia of the horseshoe crab,Limulus polyphemus (L.)

Summary The ventral cord ganglia of the horseshoe crab, Limulus polyphemus, contains six distinct cell types: three appear to be ordinary neurons and three exhibit the staining affinities of neurosecretory cells.The presumed neurosecretory cells have been termed neurosecretory cell I (NSC I), NSC II and NSC III. NSC I cells contain a colloid-like inclusion which may occur as a single small vacuole or occupy more than one-half of the cell volume. Colloid inclusions occur with greater frequency toward the periphery, although small cells of similar staining affinity occur in cords extending to the fibrous core. The histochemical tests suggest that the cytoplasm is positive for proteins, but contains no strong acidic groups which may have been derived from S-S or S-H groups. The presence of carbohydrate is also indicated.NSC II cells exhibit distinct secretory cycles. Early in the cycle the cytoplasm becomes phloxinophilic and progresses to a distinct fuchsinophilic stage. Small homogeneous irregular inclusions are found in the axon hillock during the latter stages of the cycle. Histochemical tests suggest the presence of a carbohydrate and strong acidic groups which may have been derived from S-S or S-H groups. There are small cells present which appear to be immature neurosecretory cells.NSC III cells are characterized by a perinuclear ring of cytoplasm which is stained by chrome alum hematoxylin but not by paraldehyde fuchsin. A secretory cycle may also be present in this cell type.The three cell types presumed to be ordinary neurons exhibit no particular staining affinity for the stains or tests used in this study.This study was supported in part by a grant from the Central Fund for Research of the Pennsylvania State University.     

Cytochemical studies of developmental stages during oogenesis in Culex pipiens molestus (Forsk?l). I. Lipids and carbohydrates

Lipids and carbohydrates were studied in the polytrophic ovaries of Culex pipiens molestus during oogenesis. The cytoplasm of both the oocyte and the nurse cells contains lipid structures at all stages of development--granules in the early stages and spheres in the later stages. Intranuclear lipid bodies can be demonstrated in the oocyte and in the nurse cells. After leaving the nucleus, lipids are deposited in the peripheral cytoplasm. From the third to the seventh adult phase, lipid granules are concentrated in the area of the nurse cell and oocyte junction, indicating that lipids originate in the nurse cells and are transported from these to the oocyte. The follicular epithelial cells provide the oocyte with lipid material for fatty yolk synthesis and formation of the egg envelopes. Lipids are distributed similarly to the Golgi apparatus, indicating that there is a relationship between this organelle and fat formation. In the early stages, the cytoplasm of the oocyte, the nurse cells and the follicular epithelium contains glycogen granules. In the later stages these cells also contain mucopolysaccharides. The mucopolysaccharide yolk spheres are enclosed in vacuoles, while the chorion is composed of acid mucopolysaccharides. The follicular epithelium and vitelline membrane are of a mucopolysaccharide nature. A topographical relationship exists between the Golgi apparatus and the glycogen granules, indicating that this organelle also plays a role in glycogen synthesis.     

Neurosecretory cells in the optic lobe of two aquatic coleopteran species,Dineutes indicus aube and Cybister rugulosus redt

In each optic lobe and optic peduncle of two aquatic beetles viz. Dineutes indicus and Cybister rugulosus the neurosecretory cells are observed with the help of various histochemical techniques. These cells are arranged to form a discrete group. A group in the optic lobe of both species contains about 25 to 30 neurosecretory cells. On the basis of staining properties the neurosecretory cells are classified into A and B types. These cells stain with chrome haematoxylin-phloxine and paraldehyde fuchsin, but do not stain with azan. Histochemically, the neurosecretory material is positive for proteins and shows a negative reaction for 1,2-glycols. The cells show variations in RNA contents in correlation with the state of secretory activity. Axons of the neurosecretory cell group of the optic lobe are observed directed to the optic peduncle. The axonal tract from neurosecretory cells in the optic peduncle runs towards the lateral margin of the brain.     

Histochemistry and ultrastructure of the albumen gland duct of the prosobranch gastropod Pomacea paludosa

Summary

The albumen gland duct passes through the base of the albumen gland. It consists of a single layer of cells composed of ciliated and secretory cells. Sulfated and non-sulfated acid mucopolysaccharides are secreted by the cells of the albumen gland duct. Cell resembling neurosecretory cells are also found between the ciliated and secretory cells. The secretion products probably contribute to the formation of the albumen layer which surrounds the fertilized egg.     

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.     

Histology, histochemistry, and ultrastructure of neurosecretory cells in the optic lobe of the cockroach, Periplaneta americana

In the region of the distal optic chiasma of each optic lobe of Periplaneta americana, there is a group of about 120 monopolar neurosecretory cells. These cells do not stain with paraldehyde fuchsin but remain acidophilic after oxidation. They stain red or sometimes indigo with the azan technique. Histochemically, the neurosecretory material is positive for protein and the amino acids tryptophan and arginine but negative for 1, 2-glycols and strongly acidic groups. At the ultrastructural level, the cytoplasm of the cells contain many elementary neurosecretory granules 100 to 170 nm in dia. The cells also contain well-developed Golgi bodies and endoplasmic retieulum. The axons from these cells run toward the interior of the optic lobe. In this region, axons containing dense granules (mean diameter 70 nm) and synaptic vesicles synapse onto the axons from the neurosecretory cells. The neurosecretory axons then cross over to the anterior side of the optic lobe and run towards the brain. The function of these neurosecretory cells is unknown, but they may be involved with photoperiodically controlled activity rhythms.     

Neurosecretory cells in Dysdercus similis (Hemiptera)

The neurosecretory cells of Dysdercus similis have been described. "A", "B", "C" and "D" types of neurosecretory cells are present. The "A" type of cells of the pars-intercerebralis show cyclical secretion. When these cells show secretory activity during one to three days of emergence, they have scattered granules. The cells are seen packed with clumps of neurosecretory material when they are not secreting, and this is interpreted as a storage stage. The axons of these cells supply the corpora cardiaca and some neurosecretory material also reaches the corpus allatum. The release of this neurosecretory material can be correlated both with moulting in the young stages and later with reproduction in the adults.     

Neuroendocrine control of oocyte development in Poecilocera picta (Insecta: Orthoptera)

The neuroendocrine control of oocyte development in Poecilocera picta Fab. has been described. The secretory activity of the A type of neurosecretory cells has been correlated with ovarian development. In P. picta during the first four days after emergence the neurosecretory material is seen passing down the axons but the cells are largely devoid of neurosecretory material. When the oocytes are developed the A type cells are with stored neurosecretory material.
In P. picta the mature males do not appear to accelerate the process of maturation in females. The females which are reared without males or with castrated males also mature at the same time as the females which are reared with males. The corpus allatum also enlarges and decreases at the same period. The number of resorptive bodies is much more in the females which are reared with castrated males or without males. There appears to be some correlation in the secretion of the neurosecretory material, copulation, and the appearance of resorptive bodies. In P. picta the role of the mature male is only in copulation which very likely allows the cell to synthesize and secrete and release a large amount of neurosecretory material whose discharge in the haemolymph enables a successful development of the oocyte. Corpus allatum appears to be controlled by a precursor from the brain.
Cautery of the cerebral neurosecretory cells, allatectomy and sectioning of the nerves have been done to find out the role of neurosecretory material.     

Cytochemistry of the neurosecretory cells in the crab Menippe rumphii (Fabricius, Crustacea: Brachyura)

In Menippe rumphii five types of neurosecretory cells are found in the cerebral, commissural and thoracic ganglia. Detailed cytochemical observations on the neurosecretory cells revealed that they have responded strongly to saliva resistant PAS staining. Among proteins those rich in disulfides and sulhydryl groups are observed. Greater amounts of cytoplasmic RNA are observed in the reproductive season. Considerable amounts of lipids and phospholipids are also observed in the AS cells. The cytochemical differences between the NS cells and the nonsecretory neurons are also discussed.     

The importance of disulfide bridges in human endopeptidase (enkephalinase) after proteolytic cleavage

The neutral endopeptidase (NEP) is a membrane-bound enzyme, which is solubilized by treatment with the protease, papain. Papain did not affect the apparent catalytic activity or the molecular mass of the purified human enzyme in SDS-PAGE. When NEP was treated with a reducing agent after papain digestion, it dissociated into smaller, lower molecular mass fragments. Amino acid analysis and s-carboxymethylation of the half cystine residues indicated that NEP contains four S-S bridges. We concluded that, although covalent bonds appear to be cleaved in NEP by papain, its activity and structure are sustained by S-S bridges.     

Protein and carbohydrate histochemistry in relation to the keratinization in the epidermis of Barbus sophor (Cyprinidae, Pisces)

The distribution of protein and carbohydrate constituents in the epidermis of Barbus sophor is described in order to give a better understanding of its cellular organization and physiology.
Various cytochemical techniques show the keratinized nature of the outer free margins of the polygonal cells in the most-superficial layer. These contain appreciable amounts of cysteine bound sulphydryl groups, basic proteins, protein bound NH₂ groups, ribonucleic acid and calcium and give a strong Papanicolaou's reaction. Absence of cystine bound disulphide groups suggests that the cornified layer in B. sophor is probably mechanically weak as adjacent keratin chains remain unbonded. The polygonal cells showing keratinization at the outer free margins remain metabolically active and are not sloughed off at the surface. This is in contrast to the keratinized epidermis of other teleosts so far reported in which the keratinized cells are dead and are sloughed off at the surface.
In addition to keratinization the polygonal cells undergo mucogenesis synthesizing sulphated acid mucopolysaccharides.
The presence of eosinophilic granular cells in the epidermis is interesting. The possible role of these cells in the protection of the epidermis has been discussed. The epidermis on the inner surface of the scale is very thin so it may not have much protective significance in these areas.     

Morphological and histochemical observations on the intestinal epithelium of Ascardia galli (Nematoda: Ascaridida).

The intestinal epithelium of Ascardia galli has been studied with various cytological and cytochemical techniques. It consists of large epithelial cells resting on a thick collagenous basal lamina. Their luminal surface is provided with microvilli. The intestinal cells store considerable amounts of glycogen and neutral lipids. Some intracellular granular inclusions, which stain for proteins, phospholipids and lipoproteins, are distributed throughout the cytoplasm. The brush border is composed of microvilli whereas the outer surface coat consists of saliva resistant PAS-positive material. The detailed histochemical analysis of surface material has revealed that it is composed of nonacetylated acid mucopolysaccharides rich in hyaluronic acid with carboxylate polyanions. The brush border shows intense activities of acid phosphatase and glucose-6-phosphatase, moderate of ATPase, and lipase, weak of 5'-nucleotidase. Acid phosphatase-positive intracellular structures are seen in the intestinal epithelium which form distinct aggregations.