Presence Of Rough Endoplasmic Reticulum Research Articles

Morphological and ultrastructural analysis of an important place of sexual communication of Rhodnius prolixus (Heteroptera: Reduviidae): the Metasternal Glands

Rhodnius prolixus is an important vector of Trypanosoma cruzi, the etiological agent of Chagas disease. Insect adults have a pair of Metasternal Glands (MGs) and the secretion emitted by these glands acts as sex pheromone. Recent studies have focused on the chemical composition of this pheromone, electrophysiological responses to MGs compounds and mating behavior assays. Morphological studies of these glands are still scarce. Thus, considering the relevance of MGs in the sex pheromone biosynthesis, we investigated the morphology and ultrastructure of R. prolixus MGs. The glandular apparatus presents a tubular structure containing secretory cells with canalicules that fuse with the central duct which conducts the secretion to a pear-shaped reservoir connected to the exterior by a droplet-shape orifice. The secretory cells are classified as class III, they present a well-developed rough and smooth endoplasmic reticulum. Smooth endoplasmic reticulum is a site of lipid biosynthesis that may be involved in the mevalonate pathway, a probable route of the sex pheromone biosynthesis in this insect. The presence of rough endoplasmic reticulum indicates a possible peptides/proteins secretions site which were still not characterized in MGs. Several mitochondria are scattered in the cytoplasm that may suggest a high metabolic activity. Further studies should be carried out to correlate these data with the sex pheromone biosynthesis in this vector.

Ultrastructural characterization of cells in the tibial stump of ruptured human anterior cruciate ligament, their changes and significance with duration of injury.

Fibroblasts and myofibroblasts have been known to be present in both ruptured and intact human anterior cruciate ligament (ACL), and although their relevant histology and immunochemistry have been studied in the past, ultrastructural features of these cells are largely lacking. Therefore, we aim to characterise the ultrastructural details of these cells with the help of transmission electron microscopy (TEM) and to study the changes and their significance with duration of injury. Samples from 60 ruptured human ACL undergoing surgery were obtained and categorised according to duration of injury and observed under TEM with main focus on the following ultrastructural features: cellular morphology, presence of rough endoplasmic reticulum, Golgi apparatus, lamina, myofilaments, and presence of myofibroblasts. These features were further correlated with the duration of injury and association, if any, determined using appropriate statistical analysis. A total of 54 male and 6 female patients with mean duration of the injury of 23.01 ± 26.09weeks (2-108weeks) were included in the study and categorised into five groups based on duration of injury as follows: I (< 6weeks), II (7-12weeks), III (13-20weeks), IV (21-50weeks) and V (> 50weeks). There was a significant association between the above-mentioned ultrastructural features and the duration of injury (p < 0.05) except for the presence of ovoid fibroblast cells (p = 0.53). Furthermore, number of myofibroblasts and cells with Golgi apparatus and rough endoplasmic reticulum was seen to peak at 13-20weeks following injury. We describe ultrastructural features of fibroblast of different morphology along with myofibroblasts in the ligaments following injury, the changes in which might have a potential bearing on ligament healing.

Three-Dimensional Nanostructure of an Intact Microglia Cell.

Microglia are non-neuronal cells of the myeloid lineage that invade and take up long-term residence in the brain during development (Ginhoux et al. 2010) and are increasingly implicated in neuronal maturation, homeostasis, and pathology (Bessis et al. 2007; Paolicelli et al. 2011; Li et al. 2012; Aguzzi et al. 2013, Cunningham 2013, Cunningham et al. 2013). Since the early twentieth century several methods for staining and visualizing microglia have been developed. Scientists in Ramon y Cajal's group (Achucarro 1913, Rio-Hortega 1919) pioneered these methods and their work led to the christening of microglia as the third element of the nervous system, distinct from astrocytes and neurons. More recently, a combination of imaging, genetic, and immunological tools has been used to visualize microglia in living brain (Davalos et al. 2005; Nimmerjahn et al. 2005). It was found that microglia are highly motile under resting conditions and rapidly respond to injuries (Kettenmann et al. 2011) suggesting a role for microglia in both brain homeostasis and pathology. Transmission Electron microscopy (TEM) has provided crucial complementary information on microglia morphology and physiology but until recently EM analyses have been limited to single or limited serial section studies (Tremblay et al. 2010; Paolicelli et al. 2011; Schafer et al. 2012; Tremblay et al. 2012 ; Sipe et al. 2016). TEM studies were successful in defining a set of morphological criteria for microglia: a polygonal nucleus with peripheral condensed chromatin, a relatively small cytoplasm with abundant presence of rough endoplasmic reticulum (RER), and a large volume of lysosomes and inclusions in the perikaryon. Recent advances in volumetric electron microscopy techniques allow for 3D reconstruction of large samples at nanometer-resolution, thus opening up new avenues for the understanding of cell biology and architecture in intact tissues. At the same time, correlative light and electron microscopy (CLEM) techniques have been extended to 3D brain samples to help navigate and identify critical molecular landmarks within large EM volumes (Briggman and Denk 2006; Maco et al. 2013; Blazquez-Llorca et al. 2015, Bosch et al. 2015). Here we present the first volumetric ultrastructural reconstruction of an entire mouse hippocampal microglia using serial block face scanning electron microscopy (SBEM). Using CLEM we have ensured the inclusion of both large, small, and filopodial microglia processes. Segmentation of the dataset allowed us to carry out a comprehensive inventory of microglia cell structures, including vesicles, organelles, membrane protrusions, and processes. This study provides a reference that can serve as a data mining resource for investigating microglia cell biology.

Ultrastructure of the male accessory glands of sesarmid crab, Parasesarma plicatum (Latreille, 1803)

The structure of accessory glands (AGs) in the male sesarmid crab, Parasesarma plicatum, is described by light and transmission electron microscopy. Adult males of carapace of width 1.6–2.2cm were collected from along the estuarine regions of Kanyakumari District, India. Posteriorly, the male reproductive system receives several sac-like structures, referred to as AGs. Histologically, the AG is internally lined by cuboidal epithelium and the lumen encloses eosinophilic vesicular secretions, apparently glycopreoteinaceous in nature. Ultrastructurally, the epithelium shows the signs of typical infrastructure for synthetic activity, as demonstrated by the prolific presence of rough endoplasmic reticulum, free ribosomes, and Golgi complex. The cytoplasm is manifested with electron-dense, electron-lucent, and medium-density secretory vesicles, and the mode of release into the lumen is both merocrine and apocrine. Within the lumen, these secretory vesicles coalesce and aggregate into large heterogeneous masses of varying sizes, which may play an important role post-copulation within the female duct.

Quantitative and ultrastructural changes in the haemocytes of Spodoptera littoralis (Boisd.) treated individually or in combination with Spodoptera littoralis multicapsid nucleopolyhedrovirus (SpliMNPV) and azadirachtin

The total haemocyte count (THC) and the possible ultrastructural alterations induced in the haemocytes of the fourth larval instars of the Egyptian cotton leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae), 96h post-feeding on a semi-synthetic diet, treated with the LC50 of Spodoptera littoralis multicapsid nucleopolyhedrovirus (SpliMNPV) and the LC50 of azadirachtin alone, and the LC25 of SpliMNPV combined with the LC25 of azadirachtin were studied and compared to the control. Single treatment with the virus and azadirachtin or combined treatment significantly decreased the THC compared to the control. There are five types of haemocytes in S. littoralis: prohaemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids. The most common symptoms in granulocytes and plasmatocytes, the main affected cell types, due to viral infection were the presence of virogenic stroma, peripheral dispersion of the chromatin and disappearance of the nucleoli. However, the most common symptoms in these two types of haemocytes due to treatment with azadirachtin were the presence of rough endoplasmic reticulum filled with fibrous materials, due to probably apoptosis, in their cisternae and disorganization of mitochondria (looped, vacuolated and swollen). In addition, the cytoplasm of granulocytes was vacuolated with the appearance of autophagic lysosomes, while plasmatocytes showed ruptured cell membrane and folded nuclear envelope. Combined treatment with the NPV and azadirachtin induced the same pathological changes which were recorded from individual treatment with the virus or azadirachtin to the same haemocytes. It can be concluded that the change in the THC and ultrastructure of granulocytes and plasmatocytes may affect the cellular-mediated immune response in S. littoralis. Moreover, it seems likely that mitochondria were the target site of azadirachtin, as they were affected in both granulocytes and plasmatocytes treated with azadirachtin alone or in combination with SpliMNPV.

Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow

The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium-rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n = 64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies.

Ultrastructure of differentiating oocytes and vitellogenesis in the giant freshwater prawn, Macrobrachium rosenbergii (de man)

The ultrastructure of oogenesis in Macrobrachium rosenbergii, with reference to vitellogenesis, has not been reported. We used light and electron microscopy, as well as vitellin (Vn) purification and antibody production, to study the temporal and spatial production of Vn in the ovary by immunofluorescence. Histologically, the ovary is subdivided into cone-shaped ovarian pouches with a central core containing layers of oogonia. These divide to produce oocytes that migrate outwardly and differentiate into mature oocytes. During the course of differentiation, oocytes undergo modifications, including the rearrangement of nuclear chromatin, the accumulation of ribosomes, rough endoplasmic reticulum (RER), and lipid, and the formation of secretory and yolk granules, resulting in four stages. Ultrastructurally, early previtellogenic oocytes (Oc(1)) are characterized by the accumulation of new ribosomal aggregates, translocated from the nucleus. Late previtellogenic oocytes (Oc(2)) show nuclear heterochromatin with a "clock face" pattern, the presence of RER, and three types of secretory granules. Follicular cells occupy the intercellular spaces and surround the Oc(2). Early vitellogenic oocytes (Oc(3)) are larger, with nuclei containing predominantly decondensed euchromatin, and cytoplasm with yolk and secretory granules, and few lipid droplets. Late vitellogenic oocytes (Oc(4)) are characterized by completely euchromatic nuclei, an indistinct plasma membrane, yolk platelets and secretory granules, and abundant lipid. Vitellogenin (Vg) in ovaries of M. rosenbergii consist of two main bands at MW 90 and 102 kDa. Our data indicates that Vn is present, and probably synthesized in Oc(3) and Oc(4), but there may be some undetected exogenous Vg production.

Asymptomatic Macrothrombocytopenia in a Young Pure-Bred Beagle Dog

During baseline evaluation prior to a preclinical safety study, a 10-month-old male pure-bred Beagle dog was found to have marked thrombocytopenia (6 × 10(3) platelets [PLT]/µL) associated with a mean platelet volume (MPV) of 17.9 fL. Tests for Rickettsia rickettsii, Ehrlichia canis, and Borrelia burgdorferi were negative. Buccal bleeding time was normal. Over 3 months, PLT were 4 to 141 × 10(3) PLT/µL, and MPV was 11.4 to 25.1 fL; however, PLT were <50 × 10(3) PLT/µL and MPV was >16 fL during most of this period. Antinuclear antibody (ANA) and anti-PLT antibody tests were negative. Genotyping for the presence of a beta 1-tubulin mutation demonstrated the normal wild-type gene. Treatment with prednisone resulted in normal values after only 3 days. Ultrastructure of enlarged PLT was consistent with that of immature PLT, characterized by reduced numbers of peripheral microtubules and the presence of rough endoplasmic reticulum, free ribosomes, Golgi apparatus, and a prominent canalicular system. PLT ultrastructure and glucocorticoid responsiveness supported a diagnosis of immune-mediated thrombocytopenia that was masked by the cyclic nature of PLT decreases and lack of clinical signs. Inclusion of such a dog in a preclinical safety study could result in misinterpretation of clinical pathology findings.

The salivary glands of adult female Varroa destructor (Acari: Varroidae), an ectoparasite of the honey bee, Apis mellifera (Hymenoptera: Apidae)

ABSTRACT Varroa destructor Anderson and Trueman 2000, an ectoparasite of honey bees, causes huge economic losses to apiculture annually. Its role as a vector of diseases is thought to involve the salivary glands as the terminal organs of transmission. The salivary glands are paired, oval, non-acinar organs, closely attached to the base of the gnathosoma and enveloped in a sheath of unknown embryological origin. Ultrastructurally, they are characterized by the surrounding sheath, and cells that have a strong presence of rough endoplasmic reticulum, irregular-shaped nuclei, and secretory spheres surrounded by an otherwise electron-lucent cytosol. Secretory spheres are sparse in most peripheral cells and heavily converged about the apical membranes of interior cells. Interior cells are positioned radially about the internal duct cells. The internal ducal lumen is narrow and brush-bordered. The membranes between the ducal lumen and the gland cell cytoplasm are fused and ruptured, allowing the salivary spheres to pass into the interior duct for secretion through the exterior duct. Bacterial biofilms are associated with the spheres and the rupture. Virus particles and endosymbionts are also present in the glands. The external duct remains uncharacterized, but the assumption that its embryonic origin is the same as that of the internal duct and surrounding gland cells is questioned.

Morphology of the Head Salivary and Intramandibular Glands of the Stingless BeePlebeia Emerina(Hymenoptera: Meliponini) Workers Associated with Propolis

AbstractPlebeia emerina (Friese) (Hymenoptera: Meliponini), like other stingless bees, collect large amounts of vegetal resin and store these materials, called propolis, in individualized clusters inside the nest that remain in a viscous state. The development of intramandibular and head salivary glands in P. emerina workers was studied in different life stages, aiming to relate gland functionality with the age in which they work at propolis maceration, biting the propolis clusters with the mandibles. The morphology of intramandibular and head salivary glands from newly emerged, 20–30 d old, and forager bees was analyzed. The greatest size of the head salivary glands occurred in 20–30-d-old worker bees, and the ultrastructure of this gland showed the presence of rough endoplasmic reticulum and lipid droplets. The intramandibular glands were of two types: glandular units (class 3 glands), present throughout the worker bee life span, and the secretory epithelium (class 1 glands), which hypertrophies in 20–30-d-old and forager bees. The development of the head salivary glands and the mandibular epithelium suggest that their products are added to the propolis clusters, supporting the hypothesis that they may serve in maintaining its viscous state.

Interaction of Microtubule-associated Protein-2 and p63: A NEW LINK BETWEEN MICROTUBULES AND ROUGH ENDOPLASMIC RETICULUM MEMBRANES IN NEURONS

Neurons are polarized cells presenting two distinct compartments, dendrites and an axon. Dendrites can be distinguished from the axon by the presence of rough endoplasmic reticulum (RER). The mechanism by which the structure and distribution of the RER is maintained in these cells is poorly understood. In the present study, we investigated the role of the dendritic microtubule-associated protein-2 (MAP2) in the RER membrane positioning by comparing their distribution in brain subcellular fractions and in primary hippocampal cells and by examining the MAP2-microtubule interaction with RER membranes in vitro. Subcellular fractionation of rat brain revealed a high MAP2 content in a subfraction enriched with the endoplasmic reticulum markers ribophorin and p63. Electron microscope morphometry confirmed the enrichment of this subfraction with RER membranes. In cultured hippocampal neurons, MAP2 and p63 were found to concomitantly compartmentalize to the dendritic processes during neuronal differentiation. Protein blot overlays using purified MAP2c protein revealed its interaction with p63, and immunoprecipitation experiments performed in HeLa cells showed that this interaction involves the projection domain of MAP2. In an in vitro reconstitution assay, MAP2-containing microtubules were observed to bind to RER membranes in contrast to microtubules containing tau, the axonal MAP. This binding of MAP2c microtubules was reduced when an anti-p63 antibody was added to the assay. The present results suggest that MAP2 is involved in the association of RER membranes with microtubules and thereby could participate in the differential distribution of RER membranes within a neuron.

Defence responses of white mustard, Sinapis alba, to infection with the cyst nematode Heterodera schachtii

AbstractResponses of two Sinapis alba cultivars (nematode susceptible cv. Albatros and nematode resistant cv. Maxi) to infection with the root-parasitic nematode Heterodera schachtii were investigated at the anatomical and cytological levels. In the susceptible cv. Albatros, male and female juveniles develop. Syncytia associated with female juveniles are induced in the procambium and they preferentially incorporate procambial cells abutting xylem vessels. The syncytial elements are greatly enlarged and interconnected by wide cell wall openings. Syncytia associated with male juveniles are induced in the pericycle and they incorporate primarily greatly enlarged pericyclic cells and sometimes less enlarged procambial cells. In the resistant cv. Maxi, most of juveniles remain as secondstage juveniles while some develop into males. Their syncytia are induced in and involve pericyclic cells only. Syncytia induced in cv. Maxi by males differ from those induced in susceptible cv. Albatros by less enlargement of nuclei, the presence of remnants of the central vacuole, and a marked presence of rough endoplasmic reticulum. At least three different defence responses could be discriminated in cv. Maxi: i) a hypersensitive response of vascular cylinder cells during/after nematode invasion; ii) degeneration of syncytia induced in the procambium; and iii) formation of a layer of necrotic cells between syncytium and xylem vessels.

Morphology and cytology of the nasal cavity and vomeronasal organ in juvenile and adult blind mole rats (Spalax ehrenbergi).

The blind mole rat (Spalax ehrenbergi) is a fossorial solitary rodent which exhibits extensive intraspecific aggression and uses scent markings to deter contraspecific invaders. Mole rats of different ages were captured near Tel Aviv, Israel, and sacrificed by an overdose of Xylazine hydrochloride. Olfactory epithelium sites from the nasal cavity (NC) and the vomeronasal organ (VNO) were dissected and fixed for light and electron microscopy. The mole rat's olfactory epithelium of the NC consists of several cell types, of which two types are supporting cells that comprise both microvilli and cilia but differ in staining and the presence of rough endoplasmic reticulum. The third type has no cilia. Secretory goblet cells were frequent among supporting cells of adults alone. Two types of receptor cells protrude into the NC with olfactory knobs at their apical region; one type has up to 177.6 +/- 9.4 cilia per knob plus microvilli, while the other type has only microvilli. The third type of sensory cell has no knob and contains microvilli only. The basal epithelium layer consists of short-bodied cells with round nuclei. The VNO of the mole rat is situated beneath the nasal septum, consisting of supporting, sensory, and basal cell types, with many cilia at the apical portion. At its anterior part, the VNO is connected to the NC by narrow canals. The abundance of cilia and microvilli in the mole rat olfactory cells provides the first anatomical evidence for their olfactory acuity. Such acuity is important in mole rats, compensating for their loss of vision and enabling them to detect and avoid rivals prior to potential aggressive encounters as well as to select food plants during foraging.

Morphology and cytology of the nasal cavity and vomeronasal organ in juvenile and adult blind mole rats (Spalax ehrenbergi)

The blind mole rat (Spalax ehrenbergi) is a fossorial solitary rodent which exhibits extensive intraspecific aggression and uses scent markings to deter contraspecific invaders. Mole rats of different ages were captured near Tel Aviv, Israel, and sacrificed by an overdose of Xylazine hydrochloride. Olfactory epithelium sites from the nasal cavity (NC) and the vomeronasal organ (VNO) were dissected and fixed for light and electron microscopy. The mole rat's olfactory epithelium of the NC consists of several cell types, of which two types are supporting cells that comprise both microvilli and cilia but differ in staining and the presence of rough endoplasmic reticulum. The third type has no cilia. Secretory goblet cells were frequent among supporting cells of adults alone. Two types of receptor cells protrude into the NC with olfactory knobs at their apical region; one type has up to 177.6 ± 9.4 cilia per knob plus microvilli, while the other type has only microvilli. The third type of sensory cell has no knob and contains microvilli only. The basal epithelium layer consists of short-bodied cells with round nuclei. The VNO of the mole rat is situated beneath the nasal septum, consisting of supporting, sensory, and basal cell types, with many cilia at the apical portion. At its anterior part, the VNO is connected to the NC by narrow canals. The abundance of cilia and microvilli in the mole rat olfactory cells provides the first anatomical evidence for their olfactory acuity. Such acuity is important in mole rats, compensating for their loss of vision and enabling them to detect and avoid rivals prior to potential aggressive encounters as well as to select food plants during foraging. Anat. Rec. 251:460–471, 1998. © 1998 Wiley-Liss, Inc.

Morphological characteristics of the bovine corpus luteum during the estrous cycle and pregnancy

The corpus luteum, one of the biological clocks of the estrous cycle and pregnancy, is known foremost for its production of progesterone that blocks the pituitary release of gonadotropins and prepares the uterus for a pregnancy. The cellular sources of this progesterone are the steroidogenic small and large luteal cells. Other luteal cells that are not steroidogenic, but are believed to have an important role in the function of this gland are the fibroblast, macrophages and endothelial cells. The most prominent luteal cell is the large steroidogenic cell characterized by an abundance of smooth endoplasmic reticulum and densely packed spherical mitochondria that are indicative of its contribution to most of the circulating progesterone believed to be constitutively secreted and not under the control of LH. Other distinguishing features of the large luteal cell are the presence of rough endoplasmic reticulum, prominent Golgi, and secretory granules that are indicative of endocrine cells. This cell undergoes dynamic changes across the estrous cycle and pregnancy, believed to reflect a change in progesterone and protein secretion that will eventually influence a successful pregnancy or another ovulation if pregnancy fails. The morphological characteristics of the bovine luteal cells are the focus of this review.

Biosynthesis of major platelet proteins in human blood platelets.

We studied de novo protein biosynthesis in platelets of normal adult donors and in newly formed platelets isolated from splenectomized patients with idiopathic thrombocytopenic purpura (ITP). After metabolic labelling of platelet proteins, performed with different radiolabelled amino acids or carbohydrates, a tenfold increase in incorporation of radioactivity into trichloroacetic-acid-precipitable material was obtained with ITP platelets compared to control platelets. Electron microscopic studies of ITP platelets revealed the presence of rough endoplasmic reticulum and polyribosomes, providing morphological evidence for protein synthesis. SDS-PAGE of radiolabelled ITP platelet proteins followed by autoradiography showed that [35S]methionine and [3H]leucine were incorporated into almost all Coomassie-blue-stained proteins whereas [3H]carbohydrates only labelled a few bands. Using crossed-immunoelectrophoresis we identified some of the labelled platelet compounds and demonstrated that major membrane glycoproteins (GPIb, IIb, IIIa) and alpha-granule proteins, such as fibrinogen, thrombospondin, albumin and von Willebrand factor, were synthesized in newly formed circulating platelets.

The early embryogenesis of papillary (sweat duct) ridges in primate glabrous skin: The dermatotopic map of cutaneous mechanoreceptors and dermatoglyphics

The present study documents the early innervation of the epidermis prior to the onset of differentiation of the papillary (sweat duct) ridge in glabrous digital skin of rhesus monkey embryos measuring 45, 50 and 55 mm (crown-rump) length. We observed small papillary ridges, spaced at a distance of approximately 40 microns, projecting into the dermis in the center of the distal glabrous digital pad of digits 2-5 in the 55-mm embryo. The other digital pads lacked any sign of ridge formation. A two-dimensional, approximately hexagonal grid of afferent nerves was present in the superficial dermis of all digital and palmar pads. At regular intervals of approximately 40 microns, afferent nerves ascended from the superficial dermal nerve plexus and innervated the overlying epidermis. By electron microscopy, axonal growth cones were identified contacting Merkel cells that projected several microns down into the superficial dermis in the digital pad of digit 3. Thus, the earliest wave of differentiated dorsal root ganglion neuroblasts innervates Merkel cells. Schwann cells partially encircled these growing axon tips and could be identified by the presence of rough endoplasmic reticulum and free ribosomes. The youngest embryo studied had no sign of ridge formation; however, axons ascended from the superficial dermal nerve net at 30-40-microns intervals to innervate the epidermis. We conclude that afferent nerve fibers provide a two-dimensional grid that could modulate the spacing and arrangement of the papillary or sweat duct ridges of successive digits. Such an interaction is possible between digits based on the overlapping dermatotopic maps of each rete ridge. An abnormal fingerprint could thus reflect abnormal dorsal root ganglion neuroblasts expressed through mesenchyme and epidermis.

Gut morphology of Mastotermes darwiniensis Froggatt (Isoptera : Mastotermitidae)

The digestive system of Mastotermes darwiniensis (Isoptera : Mastotermitidae) is similar to that of other lower termites. The cuticle in the proventriculus of the foregut is sculptured into 6 large dentate plates. Large setae are dispersed over the 1st and 2nd order folds and small setae are on the 3rd order folds. Epithelial cells in the foregut are of one type. The midgut cells are of one functional type, with 3 stages of the developmental cycle clearly distinguishable at any one time. There is no close association of mitochondria with the apical plasmalemma or microvilli. The epithelium rests on a basal lamina and has regularly distributed regenerative crypts. The midgut cells contain a large number of mitochondria close to the basal plasmalemma. Invaginations of the basal plasmalemma are very extensive and small tracheoles are plentiful close to the basal lamina. The presence of rough endoplasmic reticulum, most of which exists in the supranuclear cytoplasm, indicates that protein synthesis occurs in these cells. An abundance of basal plasmalemma invaginations is indicative of some fluid transport occurring across the membranes. The malpighian tubules have accumulations of mineral concretion forms as well as glycogen. Tracheoles are fairly abundant and distributed around the tubules. Epithelial cells in the region where the malpighian tubules join the midgut contain granules which are presumed to be secretory. Dentate plates around the anterior colon entrance are oriented so as not to hinder the flow of the lumen contents towards the posterior colon. Invaginations of the apical plasmalemma of the normal paunch epithelial cell suggest that they are involved in fluid or solute uptake from the lumen. The cuticle of the hindgut consists of areas with adhering bacteria and areas showing an even distribution of pores free of bacteria. The epithelial cells of the anterior colon resemble those in the paunch. The epithelial cells of the posterior colon show 3 variations: a squamous-cuboidal cell with unconstricted lumen; a cuboidal-columnar cell type found at the constriction site close to the rectum; a second squamous-cuboidal cell type constituting the junction of the colon and the rectum. This last cell type shows an abundance of cytoplasmic microtubules and a thick subcuticular zone.

Membrane Asymmetry

The components of biological membranes are asymmetrically distributed between the membrane surfaces. Proteins are absolutely asymmetrical in that every copy of a polypeptide chain has the same orientation in the membrane, and lipids are nonabsolutely asymmetrical in that almost every type of lipid is present on both sides of the bilayer, but in different and highly variable amounts. Asymmetry is maintained by lack of transmembrane diffusion. Two types of membrane proteins, called ectoproteins and endoproteins, are distinguished. Biosynthetic pathways for both types of proteins and for membrane lipids are inferred from their topography and distribution in the formed cells. Note added in proof. A cell-free system has now been developed which permits the mechanisms of membrane protein assembly to be studied (108). The membrane glycoprotein of vesicular stomatitis virus has been synthesized by wheat germ ribosomes in the presence of rough endoplasmic reticulum from pancreas. The resulting polypeptide is incorporated into the membrane, spans the lipid bilayer asymmetrically, and is glycosylated (108). The amino terminal portion of this transmembrane protein is found inside the endoplasmic reticulum vesicle, while the carboxyl terminal portion is exposed on the outer surface of the vesicle. Furthermore, addition of the glycoprotein to membranes after protein synthesis does not result in incorporation of the protein into the membrane in the manner described above (108). Consequently, protein synthesis and incorporation into the membrane must be closely coupled. Indeed, using techniques to synchronize the growth of nascent polypeptides, it has been shown (109) that no more than one-fourth of the glycoprotein chain can be made in the absence of membranes and still cross the lipid bilayer when chains are subsequently completed in the presence of membranes. These findings demonstrate directly that the extracytoplasmic portion of an ectoprotein can cross the membrane only during biosynthesis, and not after.

Ultrastructure of the Stannius corpuscles of the killifish, Fundulus heteroclitus, and its relation to calcium regulation

The ultrastructure of the corpuscles of Stannius of the euryhaline killifish Fundulus heteroclitus adapted to freshwater, seawater, and artificial calcium-deficient seawater is presented. The Stannius corpuscles of fish kept in calcium-deficient seawater appear inactive and show an accumulation of secretory granules and a relatively inactive Golgi complex. In contrast, the corpuscles of fish kept in seawater show depletion of secretory granules, hyperactivity of the Golgi complex, and dilation of the endoplasmic reticulum, indicating a response to the demand for a calcium-lowering factor in the high calcium environment. These changes are related to the difference in calcium levels of the environment and probably not to the differences in total osmotic concentration. The presence of rough endoplasmic reticulum indicates that the product of the gland may be proteinaceous.