Distribution of serotonergic neurons in the eyestalk and brain of the crab,Cancer antennarius

1. Serotonin-containing neurons were localized immunocytochemically in crab cerebral ganglia and their extensions in the eyestalk.2. Approximately 155 serotonergic cells were found in identifiable regions of the brain, the largest number being localized in the anterior cell cluster (40 reactive cells) and the bilateral anterior olfactory cell clusters (40 cells each).3. Serotonin immunoreactive cells were found in all three ganglionic divisions of the eyestalk. The medulla terminalis contains up to 15 reactive cells, of which only one occurs in the X-organ (origin of neurosecretory axons in the sinus gland nerve). The m. terminalis also contains three identifiable cells in the mediolateral border adjacent to the sinus gland nerve, of which one is a giant (up to 100 μm diameter), designated MT-1. The axon of MT-1 branches profusely after entering the m. terminalis neuropil.4. No serotonin immunoreactivity was apparent within the sinus gland, the sinus gland nerve or the organ of Bellonci.5. These findings are discussed in relation to the known serotonergic control of peptide hormone secretion by the eyestalk X-organ-sinus gland complex.     

Properties of serum high-density lipoproteins in the crab, Cancer antennarius Stimpson

Two classes of high-density lipoprotein (HDL) comprise virtually all the lipoprotein mass in female hemolymph. These lipoproteins have hydrated densities of 1.187 g/ml (HDL3) and 1.112 g/ml (HDL2). A third species (HDL1, density 1.080 g/ml) appeared in ovigerous crabs. The mean annual HDL protein concentration was 109 mg/dl of which 67% was HDL3. HDL proteins of both HDL2 and HDL3 were mostly insoluble in tetramethylurea. Three major components with apparent mol. wts of 185,000, 100,000 and 84,000 daltons were identified by gel electrophoresis in SDS. Amino acid compositions are reported. Electron microscopy indicated that the HDL are polymorphic and discoidal. Similarities in shape and differences in size of HDL3 and HDL2 particles were consistent with their lipid and protein composition. Phospholipids, mostly phosphatidylcholine, were the dominant lipid class (74%); no cholesteryl esters were detected. Palmitic and oleic acids were the major fatty acid components of esterified lipids.     

Purification and characterization of an O-acetylsialic acid-specific lectin from a marine crab Cancer antennarius

A sialic acid-binding lectin with high specificity for 9-O-acetyl- and 4-O-acetylsialic acids was purified from the hemolymph of the California coastal crab, Cancer antennarius, by affinity chromatography using bovine submaxillary mucin coupled to agarose. The binding specificity of the crab lectin distinguishes it from other known sialic acid-specific lectins from Limulus polyphemus and Limax flavus which show a broader range of specificity for sialic acids. The purified lectin is homogenous on sodium dodecyl sulfate-polyacrylamide electropherograms with a subunit molecular weight of about 36 kDa. The specificity of the lectin for O-acetylsialic acids appears to account for the fact that it agglutinates mouse, rat, rabbit, and horse erythrocytes, which contain O-acetylsialic acids on cell surface glycoconjugates, but not human monkey, sheep, goat, and chicken erythrocytes which contain only NeuAc or N-glycolylneuraminic acid (NeuGc). This conclusion was supported by the potent inhibition of hemagglutination by bovine and equine submaxillary mucins which contain 9(7,8)-O-acetyl- and 4-O-acetylsialic acids, respectively, and also by free 9-O-acetyl-N-acetylneuraminic acid (9-O-Ac-NeuAc) and 4-O-Ac-NeuAc relative to NeuAc and NeuGc. Further support for the role of O-Ac-sialic acids in hemagglutination of erythrocytes was obtained by enzymatic modification of human erythrocytes. Sialidase-treated erythrocytes were resialylated with purified sialyltransferases and various CMP-sialic acid donor substrates to contain NeuAc or NeuGc or 9-O-Ac-NeuAc in the Sia alpha 2,3Gal or Sia alpha 2,6Gal linkages. Cells resialylated to contain NeuAc or NeuGc were not agglutinated, but cells resialylated to contain 9-O-Ac-NeuAc were agglutinated with high titer, comparable to that of mice or horse erythrocytes.     

Ultrastructure of the normal human adrenal cortex]

Electron-microscopic investigations on the human adrenal cortex have been carried out in order to elucidate ultrastructural differences in the various zones. These studies indicate that certain differences in the ultrastructure of the mitochondria may be particularly useful as criteria to differentiate the three cortical zones. In this regard, the appearance of two types of mitochondria in the zona glomerulosa as well as in the zona fasciculata is of obvious interest. The probability of a relationship between the activity of the different cortical zones and the predominant presence of one form of mitochondria has been suggested. In addition, ultrastructural transformations following stress situations as well as problems dealing with the so-called 'dark and light cells' and the basal membrane are discussed.     

Ultrastructure of normal monkey liver.

The normal ultrastructure of the liver of rhesus and cynomolgus monkeys is described. A classification of type A and B periportal hepatocytes is introduced together with a discussion of the high incidence of mitochondrial abnormalities. It is suggested that the rhesus monkey is more suitable as an experimental model than the cynomolgus since its liver ultrastructure is less variable.     

DNA reassociation kinetics and chromosome structure in the crabs Cancer borealis and Libinia emarginata

DNA reassociation kinetics have been partly elucidated for the higher crabs C. borealis and L. emarginata, using calf thymus DNA as a standard. These crabs contain no detectable repeated DNA in the approximate multiplicity frequency range 2-100 copies, which is unusual for invertebrate DNAs. Each species contains a component renaturing at an intermediate rate, and also a very rapidly renaturing fraction. The very rapidly renaturing fraction is considerably larger than the cesium chloride-resolvable satellites of each species. The fraction reassociating at an intermediate rate includes sequences with a reiteration frequency of up to 9.0 X 10(4) copies. This is unusually high for invertebrate DNAs. The nearly exact correlation between kinetic complexity and independently determined haploid genome size leads to the conclusion that the most slowly renaturing sequences of both crab species are present only once per haploid genome. Therefore the chromatids of these species are uninemic structures, and there has been no detectable occurrence of polyploid speciation in the recent evolutionary history of either species.     

Uptake of high-density lipoprotein by Y-organs of the crab,Cancer antennarius. I. characterization in vitro and effects of stimulators and inhibitors

Cholesterol is the obligate precursor for ecdysteroid hormone synthesis by the ecdysial glands (Y-organs) in crustaceans, and all cholesterol in the hemolymph is bound to high-density lipoprotein (HDL). The mechanism was studied of how Y-organ cells acquire cholesterol. Y-organ segments were incubated with HDL isolated from hemolymph and labeled with ¹²⁵I. After incubation, tissue was homogenized in acid to determine radioactivity in acid-precipitable (cell associated, intact) HDL and in acid-soluble (degraded) HDL. Both HDL uptake and degradation showed saturation kinetics. At saturation most of the total counts represented degraded HDL; by 3 h, degradation was 80%. Rates of HDL uptake and breakdown were higher in Y-organs from de-eyestalked crabs (deprived thereby of molt-inhibiting hormone, MIH) than in glands from intact crabs. Both parameters were depressed by inhibitors of glycolysis and oxidative phosphorylation dose dependently and by low temperature. HDL uptake also was depressed by cAMP added to the medium experimentally or through efflux from the tissue during incubation. These results indicate a mechanism for HDL uptake that entails receptor-mediated, energy-dependent endocytosis of the entire HDL-cholesterol complex. Also the results suggest that HDL uptake and degradation are mediated by cAMP and depressed by an eyestalk factor, presumably MIH. © 1995 Wiley-Liss, Inc.     

Ultrastructure of the nucleus of human sertoli cells in normal and pathological testes

Cell and Tissue Research - The ultrastructure of the nucleus of human Sertoli cells has been studied in testicular samples from normal adults and patients with hypospermatogenesis or absence of the...     

Ultrastructure of surface epithelial cells of the normal human rectum

Luminal surface epithelial cells, excluding a few endocrine cells of the normal human rectum, were studied electron microscopically and 5 types of cells were recognized with special reference to some structure containing mucous substances. Principal-1 cells showing few tiny vesicles and Principal-2 cells containing some tiny vesicles seemed to belong to the absorptive cell group. Vesicle cells having numerous tiny vesicles, and Columnar mucous cells accompanied by numerous tiny vesicles and some round or oval mucous vacuoles, seemed to be labelled as of the secretory cell group. The common features of the epithelial columnar cells, except for the Goblet cell, were columnar shape, microvilli whose length and density had considerable variation, glycocalyceal bodies around the microvilli, and thick surface coat. Goblet cells were characterized by a goblet shape which was expanded by numerous mucous droplets. It is of special interest that 4 different types of columnar epithelial cells are recognized on the luminal surface of the normal human rectum, and that Vesicle cells and Columnar mucous cells are first observed on the luminal surface of the large intestine. Similar epithelial cells have only been reported in the crypt of the large intestine and not on the luminal surface.     

Respiratory and digestive responses of postprandial Dungeness crabs, Cancer magister, and blue crabs, Callinectes sapidus, during hyposaline exposure

Respiratory responses and gastric processing were examined during hyposaline exposure in two crab species of differing osmoregulatory ability. The efficient osmoregulator, Callinectes sapidus, displayed an immediate increase in oxygen uptake when exposed to low salinity in isolation. In contrast, the weak osmoregulator, Cancer magister, showed no change in oxygen uptake upon acute exposure (<6 h), but slight increases in oxygen uptake tended to occur over longer time scales (12–24 h). These changes were likely attributable to an increase in avoidance activity after 6 h hyposaline exposure. Following feeding in 100% SW, oxygen uptake doubled for both species and remained elevated for 15 h. When postprandial crabs were exposed to low salinities, C. sapidus were able to sum the demands of osmoregulation and digestion. Thus, gastric processes continued unabated in low salinity. Conversely, postprandial C. magister prioritized responses to low salinity over those of digestion, resulting in a decrease in oxygen uptake when exposed to low salinity. This decrease in oxygen uptake corresponded to a reduction in the rate of contraction of the pyloric stomach and a subsequent doubling of gastric evacuation time. The current study is one of the few to illustrate how summation or prioritization of competing physiological systems is manifested in digestive processes.     

Ultrastructure and relationships of spermatozoa of the freshwater crabs Potamon fluviatile and Potamon ibericum (Crustacea, Brachyura, Potamidae)

The spermatozoa of Potamon fluviatile and P. ibericum are virtually indistinguishable and do not support separate subgeneric rank ( Potamon and Pontipotamon , respectively). Synapomorphic with the spermatozoa of the South African freshwater crab Potamonautes perlatus Sidneyi are the elongation of the two centrioles and disposition of the centrioles almost parallel to each other, unknown elsewhere in the Brachyura, and reduction of the thickened ring (homoplasic with grapsids and gecarcinids). Other, probably synapomorphic, similarities of Potamon and Potamonautes include the wide inner acrosome zone, absence of a definite acrosome ray zone (homoplasic in other families) and the cleistospermial spermatophores. Further similarities, of questionable polarity, are the simple, not multilaminar, nuclear membrane and the tendency of the nuclear arms to wrap around the nucleus. Differences of Potamon from Potamonautes , which possibly support their present generic separation and give weak support to their former separate familial placement in the Potamidae and Potamonautidae respectively, are perforation of the operculum and the weak, rather than strong, development of a periopercular rim. Absence in Potamon and Potamonautes of an accessory opercular ring and a xanthid ring separates them from xanthids. No close affinities with other heterotreme families are seen but their assignment to the Helerotremata is not in doubt. Their spermatozoa lack two of the distinctive features of thoracotreme spermatozoa (apical button on the operculum and concentric lamellation of the outer acrosome zone). No clear correlates of spermatozoon structure with a freshwater existence are recognizable but reduction of the thickened ring possibly relates to peculiarities of the acrosome reaction. However, the production of spermatophores with single spermatozoa (cleistospermia) is possibly a device to prevent polyspermy and wastage of the small number of lecithotrophic eggs produced in potamids.     

Phylogenetics of Cancer crabs (Crustacea: Decapoda: Brachyura).

We used morphological, mitochondrial DNA sequence, paleontological, and biogeographical information to examine the evolutionary history of crabs of the genus Cancer. Phylogenies inferred from adult morphology and DNA sequence of the cytochrome oxidase I (COI) gene were each well resolved and well supported, but differed substantially in topology. Four lines of evidence suggested that the COI data set accurately reflected Cancer phylogeny: (1) in the phylogeny inferred from morphological data, each Atlantic species was sister taxon to an ecologically similar Pacific species, suggesting convergence in morphology; (2) a single trans-Arctic dispersal event, as indicated by the phylogeny inferred from COI, is more parsimonious than two such dispersal events, as inferred from morphology; (3) test and application of a maximum likelihood molecular clock to the COI data yielded estimates of origin and speciation times that fit well with the fossil record; and (4) the tree inferred from the combined COI and morphology data was closely similar to the trees inferred from COI, although notably less well supported by the bootstrap. The phylogeny inferred from maximum likelihood analysis of COI suggested that Cancer originated in the North Pacific in the early Miocene, that the Atlantic species arose from a North Pacific ancestor, and that Cancer crabs invaded the Atlantic from the North Pacific 6-12 mya. This inferred invasion time is notably prior to most estimates of the date of submergence of the Bering Strait and the trans-Arctic interchange, but it agrees with fossil evidence placing at least one Cancer species in the Atlantic about 8 mya.     

Ultrastructure of human amnion and amniotic plaques of normal pregnancy

Summary The fine structure of amniotic and amniotic-plaque epithelia has been studied from normal term pregnancies. The columnar/cuboidal amniotic epithelial cells usually have apical or central nuclei, some free ribosomes, patches of granular endoplasmic reticulum, juxtanuclear Golgi complexes, rod-shaped mitochondria, lipid droplets and some glycogen granules. They have short, blunt microvilli which frequently branch and bathe in the amniotic fluid. The lateral plasma membranes enclose tortuous intercellular spaces which are always interrupted by variously folded processes and desmosomes. The epithelial cells rest on a basal lamina and exhibit highly folded basal processes. The amniotic epithelial cells are neither distinctly Golgi and fibrillar types nor light and dark in appearance.Amnion from near the umbilical cord contains many microscopic and several large plaques. Similar structures are not found on the reflected amnion. The microscopic plaques are whitish and translucent, whereas the large ones are opaque. The large plaques vary between 1–3 mm in diameter, and are over 15 cell layers thick. Each large plaque has a main central region and edges continuous with either the microscopic plaque or the simple amniotic epithelium. The main region shows four zones, namely, stratum basale, stratum spinosum, stratum granulosum and stratum corneum. Such zones are not distinct at the edges. The fine structure of basal cells compares with the amniotic epithelial cells, but the cells of spinosum and granulosum layers possess variable amounts of tonofibrils, keratohyalin granules, free ribosomes and other cytoplasmic organelles and inclusions. The corneum cells are keratinized and are frequently separated by intercellular spaces. They slough into the amniotic cavity singly or as a sheet, and contribute towards the composition of the amniotic fluid. The plaques are of amniotic origin, and are not formed by adhesion of either squamous cells or fetal skin cells (masses of keratinized squames). The present observations suggest that the occurrence of amniotic plaques is normal. The presence of plaques may not be necessarily associated with fetal abnormality. However, increase in numbers of plaques may be caused by conditions of fluid imbalance. The homology and significance of plaques in eutherian mammals have been discussed.This research was supported by USPHS Grant AM-11376 and NIH Grant 69-2136.     

The cuticle of the crabs Cancer pagurus L. and Carcinus maenas (L.)

Fine closely-packed parallel fibres pass obliquely, at an angle of about 10° to the horizontal, through the cuticles of Cancer and Carcinus . They lie on axes parallel to the four faces of an obtuse pyramid, and have no counterpart in the model proposed by Bouligand (1965, 1971, 1972).
Replication of vertically broken surfaces of cuticle on cellulose acetate film shows that the laminae of the cuticle are discrete structural entities which preserve their identity around the angle formed by two vertical faces meeting at right-angles. This does not conform to the requirements of the Bouligand model.     

Ultrastructure and peroxidase cytochemistry of normal human leukocytes at birth

At birth, differential and white blood cell counts of normal newborn infants are strikingly different from those of adults in that the number of leukocytes is increased and immature cells course through the circulation. In this study, our intent was to examine normal neonatal cord blood by electron microscopy and peroxidase cytochemistry to determine whether any detectable differences exist in the leukocytes of neonatal and adult blood. This investigation was undertaken because newborn infants have an increased susceptibility to infection, and alterations in phagocyte function have been implicated as the cause. Cord blood was found to contain mature leukocytes of all kinds, similar in ultrastructure and peroxidase localization to those of adults. Moreover, as indicated earlier by light microscopy, immature forms (normally found only in adult bone marrow) were present in the blood of newborns. We found that nearly all cell lines were represented in the neonatal circulation by such developmental forms as promyelocytes, myelocytes, promonocytes, erythroblasts, megakaryocytes, rare unidentifiable blasts, and dividing cells—all resembling their counterparts in adult bone marrow. With the techniques used here, neonatal leukocytes were similar to those of the adult in ultrastructure and peroxidase localization, although some had been mobilized into the blood in a remarkably immature state. This study, the first of its kind, will serve as a helpful background for future investigations of acquired, genetic or neoplastic leukocyte abnormalities which may be discovered at birth.