Developmental Aspects of Ultrastructure, Histochemistry and Receptivity of the Stigma of Nicotiana sylvestris

The bilobed papillate stigma of Nicotiana sylvestris Speg. andComes, is covered at maturity with a copious exudate containinglipid, protein and carbohydrate. The stigma is receptive fromthe very early stage of development and it also stains positivelyfor esterase activity. The stigma has three distinct zones:an epidermis with papillae; a subepidermal secretory zone; anda parenchymatous ground tissue. The behaviour of the cells ofthese three zones has been followed from 6 d before anthesisto one day after anthesis and pollination. The cells of theepidermis and the secretory zone stain intensively for lipids,proteins and carbohydrates in the initial stages. The secretoryzone develops large intercellular spaces containing heterogenoussecretory products which also stain positively for the aforesaidthree compounds. At maturity the secretory products are releasedto the surface through gaps formed in the epidermis by cellseparation. The main secretion of the stigma is produced bythe cells of the secretory zone. Less secretion is derived fromthe stigmatic papillae. Some amount of secretion is also releasedfrom the stylar transmitting tissue adjoining the stigma. Theglandular cells of the stigma contain numerous plastids, mitochondria,ribosomes, ER, cytoplasmic lipid droplets and some dictyosomes.The plastids and the vacuoles in the secretory cells of thestigma have a lot of electron dense (osmiophilic) inclusionsrespectively in the initial and later stages of development.The former are probably involved in the production of thesematerials. It is suggested that the proteins are directly secretedby rough ER compartments whereas smooth ER is involved in thesynthesis of lipidic materials. The carbohydrate moiety of theexudate is released by the eccrine mode (sugar mono- and dimers)with some addition of polymers by disintegration of the middlelamellae. The means by which the lipidic and osmiophilic materialis extruded remains unclear. Nicotiana sylvestris, stigma receptivity, organization, stigmatic secretory system, stigmatic exudate     

Cells--Analogs of the Fat Body in the Parasitic Mite Myobia murismusculi (Schrank) (Trombidiformes: Myobiidae)

In body cavity of Myobia murismusculi females the cells with morphology similar to that in elements of fat body of other arthropods were found. They are characterized by large size, hypertrophied GER, and few secretory granules of two types. No such cells were revealed in males of the same mite species. Fat body in females at the nymphal stage is presented by poorly differentiated cells without secretory inclusions. A probable participation of the described tissue in vitellogenesis as well as the reason for its absence in other species of trombidiform mites are discussed.     

Ultrastructure of the annual cycle of female sperm storage in spermathecae of the torrent salamander, Rhyacotriton variegatus (Amphibia: Rhyacotritonidae)

This study is the first report on the ultrastructure of the sperm storage glands (spermathecae) in the salamander Rhyacotriton variegatus. The population studied is associated with cold-water, rocky streams of the redwood (Sequoia) zone in northern California. Males possess sperm in their vasa deferentia and undergo spermiation throughout the year, but mating is seasonal. Most females with large, vitellogenic follicles (2.0-3.9 mm mean dia.) collected from February-June contain sperm in their spermathecae, although some females with large follicles lack sperm. Other mature-size females collected during this period have small ovarian follicles (0.9-1.2 mm mean dia.) and lack stored sperm. All females collected from September-November have small follicles (0.6-1.6 mm mean dia.) and lack sperm, except in one instance in which a female collected in November had a small amount of degraded sperm, apparently retained from the previous breeding season. The spermathecae consist of simple tubulo-alveolar glands in which the neck tubules produce a mucoid secretory product, and the distal bulbs, where sperm are stored, contain secretory vacuoles of uniform density that stain positively for glycosaminoglycans. In specimens containing sperm, some bulbs have abundant sperm and others lack sperm, but the ultrastructure is similar in both conditions. The acini contain columnar epithelial cells with wide intercellular canaliculi, and a merocrine process releases the secretion. Spermiophagy occurs. In specimens from spring and summer with small ovarian follicles, the neck tubules are similar to those of breeding females, but the distal bulbs are reduced to cords of cells lacking a discernible lumen. Secretory activity in the distal bulbs is initiated in the fall. Spermathecae of R. variegatus are most similar to those of a stream-dwelling plethodontid, Eurycea cirrigera.     

Influence of continuous light and darkness on the secretory pinealocytes of<Emphasis Type="Italic">Heteropneustes fossilis</Emphasis>

In an earlier study onHeteropneustes fossilis, evidence of secretory activity in the pinealocytes had been demonstrated at the electron microscopic (EM) level and it was found to exist in two phases: a secretory phase (light cells) and a storage phase (dark cells). In the present investigation,H. fossilis was subjected to artificial photoperiods of continuous illumination and continuous darkness for a period of ten days and the effect on the secretory pinealocytes was studied at the EM level. Marked results were observed within the short period of ten days emphasizing the role of environmental photoperiod on the secretory activity of the pinealocytes. During continuous illuminated phase, both light and dark cells were observed: the light cells showed intense secretory activity and dark cells a storage one. During the dark phase both types of cells were present but in different metabolic states and neither of the cells demonstrated synthetic nor storage activity. Light cells were metabolically active but not secretory active and dark cells showed a necrotic condition. Phagocytotic activity of the dark cells was also seen. Intense neural activity was also observed during exposure to both the artificial photoperiods. The results highlight the role of light on the secretory activities of the pinealocytes of the catfish pineal organ.     

Subcellular localization of gonadotrophic hormones LH and FSH in frog adenohypophysis using double-staining immunocytochemistry

We applied double post-embedding immunocytochemical methods using specific antibodies against bullfrog (Rana catesbeiana) luteinizing hormone (LH) and follicle-stimulating hormone (FSH) with immunogold staining (5- and 20-nm particles) to determine the subcellular localization of both gonadotropins and to observe their immunostaining patterns in anterior pituitary of the frog Rana pipiens. Results showed that individual gonadotrophs may store either one or both gonadotropins in a given secretory granule and in large globules (lysosomes?). Most gonadotrophs (50-88%) contain both hormones; 12-50% contain only FSH, and only a few (0-7%) contain LH alone. Individual secretory granules, even in cells that contain both hormones, may contain only one or both gonadotropin molecules. Evaluation of the percentage of monohormonal and multihormonal secretory granules revealed that multihormonal secretory granules were the most numerous and that LH monohormonal secretory granules were the least numerous. These results indicate that cellular storage of gonadotropin in amphibian pituitary is similar to that described for mammals, where a single cell type containing both gonadotropins predominates. Variability in hormone content both of cells and of granules in all individuals is consistent with the hypothesis that frog pituitary possesses a single multipotential gonadotroph.     

Glandular Trichomes of Fagonia L. (Zygophyllaceae) Species: Structure, Development and Secreted Materials

The glandular trichomes ofFagoniaconsist of one secretory celland a multicellular stalk, which develops by division, elongationand elevation of epidermal cells. The latter become seperatedfrom the mesophyll and a subepidermal chamber is formed. Thelength of the stalk, which differs in the various species orvarieties is determined by the number of cell divisions and/or the extent of cell elongation. Although the basic morphologyand development of the trichomes of the species and varietiesexamined are similar, two types of mature trichomes can be distinguished:one occurs in the two examined varieties ofF. mollisand thesecond inF. glutinosaandF. arabica. The secretory cells of thesecond type possess a very thick wall and bear a porous cupuleon their top. Histochemical tests revealed that the sticky substancesecreted by the secretory cells contains mainly polysaccharidesand lipophilic compounds. The secreted material exhibits autofluorescence.InF. mollisvar.hispidachanges in the amount and shape of thefluorescent material inside the secretory cell, during the courseof development, have been observed. The contribution of theglandular trichomes inFagoniaspecies to survival in hot desertconditions is discussed. Fagonia; glandular trichomes; subepidermal chamber; secreted material; adaptation to desert conditions; stalk; fluorescence     

A PATTERN OF EPIDERMAL CELL MIGRATION DURING WOUND HEALING

Epidermal repair during wound healing is under investigation at both the light and electron microscopic levels. Suction-induced subepidermal blisters have been employed to produce two complementary model wound healing systems. These two model systems are: (a) intact subepidermal blisters, and (b) opened subepidermal blisters (the blister roof was removed immediately after induction, leaving an open wound). From these studies a pattern of movement for epidermal cells in wound healing is proposed. This pattern of movement is the same for both model systems. Epidermal cells appear to move by rolling or sliding over one another. Fine fibers oriented in the cortical cytoplasm may play an important role in the movement of these epidermal cells. Also instrumental in mediating this movement are intercellular junctions (desmosomes) and a firm attachment to a substrate through hemidesmosomes. In the intact subepidermal blisters hemidesmosomal attachment is made to a continuous and homogeneous substrate, the retained basal lamina. In the opened subepidermal blisters contact of epidermal cells is made to a discontinuous substrate composed of sporadic areas of fibrin and underlying mesenchymal cells.     

Immunocytochemical study of the GH cells in the anterior pituitary gland of human fetus II. Anencephalic fetus

In order to elucidate the effects of hypothalamic regulation on the morphology of GH cells, light and electron microscopic immunocytochemical examinations were carried out comparing GH cells in the anterior pituitary gland of anencephalic fetus with those of normal fetuses. Three types of GH cells were identified in the anterior pituitary gland of anencephalic fetus as well as in the normal fetus. Type-I is a small, round cell containing a few small secretory granules. Type-III is a large, polygonal cell with numerous large secretory granules. Type-II is a polygonal cell with medium-sized secretory granules. The Type-II GH cell was predominant in both anencephalic and normal fetuses. The most striking difference between anencephalic and normal fetuses was the presence of atypical forms of the Type II cell. These were polygonal cells containing secretory granules, which were either immunopositive or immunonegative to anti-human GH (anti-hGH) serum. Furthermore, two other types of GH cells were identified. The somatomammotroph (SM cell) contained GH and PRL in different granules within the same cell. Also, a different type of the GH cell was noted containing two varieties of secretory granules; one was immunolabeled only with anti-hGH and the other was not immunolabeled to either anti-hGH or anti-human PRL (anti-hPRL). From these results, we suggest that an absence of hypothalamic regulation in the anencehpalic does not seriously modify GH cell morphology but induces an altered GH storage pattern in some of the cells.     

Sex peptide receptor is required for the release of stored sperm by mated Drosophila melanogaster females

The storage of sperm in mated females is important for efficient reproduction. After sperm are transferred to females during mating, they need to reach and enter into the site(s) of storage, be maintained viably within storage, and ultimately be released from storage to fertilize eggs. Perturbation of these events can have drastic consequences on fertility. In Drosophila melanogaster, females store sperm for up to 2 weeks after a single mating. For sperm to be released normally from storage, Drosophila females need to receive the seminal fluid protein (SFP) sex peptide (SP) during mating. SP, which binds to sperm in storage, signals through the sex peptide receptor (SPR) to elicit two other effects on mated females: the persistence of egg laying and a reduction in sexual receptivity. However, it is not known whether SPR is also needed to mediate SP’s effect on sperm release. By phenotypic analysis of flies deleted for SPR, and of flies knocked down for SPR, ubiquitously or in specific tissues, we show that SPR is required to mediate SP’s effects on sperm release from storage. We show that SPR expression in ppk⁺ neurons is needed for proper sperm release; these neurons include those that mediate SP’s effect on receptivity and egg laying. However, we find that SPR is also needed in the spermathecal secretory cells of the female reproductive tract for efficient sperm release. Thus, SPR expression is necessary in both the nervous system and in female reproductive tract cells to mediate the release of stored sperm.     

Repugnatorial glands with associated striated muscle and sensory cells inMelibe leonina (Mollusca,Nudibranchia)

Summary The nudibranch molluscMelibe leonina is repellant to some seastars and fish, but not to crab predators. Distinctive subepidermal glands, which are particularly abundant within the cerata and dorsal wall of the oral hood, can be easily seen within the translucent body wall of this slug. The glands can be individually discharged by touching the overlying epidermis and the collected secretion is repugnant to seasters. Ultrastructural analysis reveals that each repugnatorial gland is composed of two types of secretory cell encased by cross-striated muscle. Storage of secretory product within a single, large, intracellular vacuole and cytological features of the muscle investment are consistent with a rapid, all-or-none secretion event, as observed in live tissue. The secretory cells empty into a short duct that opens by a pore. The pore is surrounded by ciliated sensory cells that probably act as mechanoreceptors triggering gland discharge. I argue that one of the two secretory cells releases defensive chemical, whereas the second may produce a protein-rich mucin.     

Cold storage of the predatory mite Neoseiulus californicus is improved by pre-storage feeding on the diapausing spider mite Tetranychus urticae

Low air temperature accompanied with high humidity is effective for long-term cold storage of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae). To further improve this storage method, we investigated the effect of pre-storage nutrition on survival during storage and on post-storage quality in terms of survival, oviposition, and progeny viability. The predatory mite was fed from the egg to adult stage on the diapausing two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), non-diapausing spider mites, or Japanese pear pollen, Pyrus pyrifolia Nakai. Newly emerged N. californicus adult females and males were mated, and then both were stored at 7.5 °C and a vapor pressure deficit of 0.0 kPa for up to 75 days. Survival during storage and post-storage quality was significantly better with the diapausing spider mite diet than with the other diets. No effects on the survival or sex ratio of the progeny of the stored adults were observed, regardless of diet or storage duration. Providing diapausing spider mites as a pre-storage diet therefore significantly improves the long-term storage of N. californicus. We discuss the possibility that ingestion of the cryoprotectants, antioxidants, and energy reserves that are present in rich amounts in diapausing spider mites mitigates chilling injury.     

Ultrastructure and functional versatility of hirudinean botryoidal tissue

In leeches, the botryoidal tissue is composed of two different cell types--granular botryoidal cells and flattened endothelial-like cells--localized in the loose connective tissue between the gut and the body wall sac. We have observed that the botryoidal tissue undergoes functional and structural modifications in response to the different needs arising during the life-cycle of the animal. In healthy, untreated leeches, botryoidal cells are organized in cords or clusters, sometimes surrounding few, small lacunae. Conversely, in wounded animals we have observed the transition of the botryoidal tissue from cluster/cord-like structures to a hollow/tubular architecture, typical of pre-vascular structures. We have documented in botryoidal cell cytoplasm the presence of large calcium storage. Moreover, the cytoplasm of botryoidal cells was filled with granules of different form and size, containing iron or melanin, as tested by classic histochemical methods. The presence of elements like iron and calcium was confirmed by the well-established EDS analysis. In response to a surgical wound, botryoidal tissue cells changed their shape and formed new capillary vessels. Concurrently, botryoidal cells secreted iron from cytoplasmic granules into the new cavity: this secretory activity appeared to be related to intracellular calcium fluctuations. At the end of the angiogenic process, botryoidal cells lost their contact with the basal lamina and moved freely in the circulating fluid towards the lesioned area. Interestingly, circulating botryoidal cells were found to carry melanin in the wounded area. This function is probably involved in defense processes. Thus, we have shown that stimulated botryoidal tissue displays a variety of striking structural, secretory and defensive activities.     

Lip morphology and ultrastructure of osmophores in Cyclopogon (Orchidaceae) reveal a degree of morphological differentiation among species

Floral fragrances play an important role in pollinator attraction; they serve as signals for primary rewards such as nectar. The presence of osmophores (a specialized glandular tissue that produces fragrance) in the terrestrial orchid Cyclopogon has only been described in a single species. There are fragrant and odorless species within this genus, but no data on the similarities or differences between them has been published. Here, we present new data on the lip morphology and ultrastructure from eight Cyclopogon species to better understand the mechanisms of pollinator attraction of this group. To achieve this goal, we used light, scanning, and transmission electron microscopy. Five out of eight species presented globular trichomes associated with fragrance production on the abaxial side of the labellum. Together with subepidermal cells, they compose the secretory tissue. Despite the absence of globular trichomes, C. apricus also presented cells associated with fragrance production. Lipid droplets, plastids containing plastoglobuli, and starch were indicators of secretory activity present in these species. This same pattern of fragrance production and release, associated with flower morphology, may result in the restriction of pollination by halictid bees.

     

The effect of interspecific mating on sex ratios in the twospotted spider mite and the Banks grass mite (Acarina: Tetranychidae)

Mixed populations of the twospotted spider mite (TSM),Tetranychus urticae (Koch), and the Banks grass mite (BGM),Oligonychus pratensis (Banks), occur on corn and sorghum plants in late summer in the Great Plains. Interspecific matings between these arrhenotokous species occur readily in the laboratory but yield no female offspring. The effect of interspecific mating on female: male sex ratios was measured by examining the F₁ progeny of females that mated with both heterospecific and conspecific males in no-choice situations. TSM females that mated first with BGM males and then with TSM males produced a smaller percentage of female offspring than TSM females that mated only with TSM males (43.1±5.8 and 78.9±2.8% females, respectively). Similarly, BGM females mated with heterospecific males and then with conspecific males produced fewer female offspring than females mated only with BGM males (55.7±5.2 and 77.5±2.5%, respectively). Lower female: male sex ratios were produced also by BGM females that mated with TSM males after first mating with conspecifics (62.4±3.4%). In mixed populations containing males of both species, females also produced lower female: male sex ratios, but these ratios were not as low as expected based on mating propensities and progeny sex ratios observed in no-choice tests. These data suggest that interspecific mating may substantially reduce female fitness in both mite species by reducing the output of female offspring, but in mixed populations this effect is mitigated by unidentified behavioral mechanisms.     

A revised diagnosis of the feather mite genus Magimelia Gaud, 1961 (Pterolichoidea: Pterolichidae: Magimeliinae) and the description of three new species

Three new mite species of the genus Magimelia (Astigmata: Pterolichidae) are described from the plumage of various lapwings (Charadriidae: Vanellinae): M. breviloba n. sp. from Vanellus miles miles; M. thailandica n. sp. from V. indicus (type-host), V. duvaucelii and V. tricolor; and M. chilensis n. sp. from V. chilensis. An extended host range for M. dolichosikya Gaud, 1961 is given. A revised diagnosis of the genus and a key to known species are presented.     

Microstructure of the prothoracic mycangia in the ambrosia beetle Platypus koryoensis (Coleoptera: Curculionidae: Platypodinae)

Platypus koryoensis (Murayama) (Coleoptera: Curculionidae: Platypodinae) is a minute ambrosia beetle that can cause significant damage to live oak trees (Quercus mongolica) along with its symbiotic pathogenic fungus (Raffaelea sp.). Although both sexes of the beetle have a number of cuticular depressions for carrying microorganisms, only the females have mycangia that are typical in size on the pronotum. The mycangial cavities are filled with a mass of amorphous substances including yeast-like spores. In addition, mycangial cavities are equipped with cuticular pores which release substances to support symbiotic microorganisms during storage. Small depressions of both sexes can be subdivided into two clusters according to their relative sizes and locations. Although they have a typical cuticular peg (basiconic sensillum), they also share a common morphological characteristic with the typical mycangium of female: a round outlet and an invaginated reservoir containing a mass of secretory substances. Accordingly, our fine structural data suggest that both the female mycangia and the clusters of small cuticular depressions in both sexes also have a mycangial function in common regardless of their relative size and location.     

Precious essences: female secretions promote sperm storage in Drosophila

Sperm that females receive during mating are stored in special places in the females' reproductive tracts. These storage sites serve to support and retain the sperm, maintaining the sperms' motility and, in mammals, permitting final sperm-maturation. The molecules that attract sperm to these sites and mediate what happens to them there have remained elusive. New research, using elegant genetic tools in Drosophila, shows that secretory cells associated with a sperm storage organ are important in sperm-supportive functions. When females lack function of these cells, they do not store sperm, or the sperm that they do store lose motility. Intriguingly, these effects influence gametes beyond the secretory cells' immediate vicinity. Loss of these cells eliminates the motility of sperm stored elsewhere in the reproductive tract and prevents the movement of eggs through the tract to exit the female. As a result of the latter problem, fertilized eggs hatch inside female flies that lack these secretory cells: instead of laying eggs, these females can "give birth" to live offspring. Because the cellular source of these gamete-regulating substances is now known, future studies can identify the specific molecules and mechanisms by which a female attracts sperm into storage and regulates the movement of sperm and eggs within her reproductive tract. It will be fascinating to determine how these molecules and mechanisms maintain gametes in active and viable forms and how evolution can modulate this to result in diverse reproductive strategies. Identification of these molecules also has potential practical implications for strategies to regulate the reproduction of insects of medical or agricultural importance.     

Anatomy and ultrastructure of the salivary (prosomal) glands in unfed water mite larvae Piona carnea (C.L. Koch, 1836) (Acariformes: Pionidae)

Anatomy and ultrastructure of prosomal salivary glands in the unfed water mite larvae Piona carnea (C.L. Koch, 1836) were examined using serial semi-thin sections and transmission electron microscopy. Three pairs of alveolar salivary glands shown are termed lateral, ventro-lateral and medial in accordance with their spatial position. These glands belong to the podocephalic system and are situated on the common salivary duct from back to forth in the above mentioned sequence. The arrangement of the medial glands is unusual because they are situated one after another on the medial (axial) body line, therefore they are termed anterior and posterior medial glands. The secretory duct of the anterior medial gland mostly turns right, and the duct of the posterior gland turns left. The salivary glands are located in the body cavity partly inside the gnathosoma and in the idiosoma in front of the brain (synganglion). Each gland is represented by a single acinus (alveolus) and is composed of several cone shaped secretory cells arranged around the large central (intra-acinar) cavity with the secretory duct base. The cells of all glands are filled with secretory vesicles of different electron density. The remaining cell volume is occupied by elements of rough endoplasmic reticulum, and the membrane enveloping vesicles may have ribosomes on its external surface. Large nuclei provided with large nucleoli occupy the basal cell zones. The pronounced development of the prosomal salivary glands indicates their important role in extra-oral digestion of water mite larvae.     

Histological events leading to somatic embryo formation in cultured petioles of alfalfa

Summary An optimum 10-day exposure of petioles of alfalfa [Medicago sativa ssp.falcata (L.) Arcangeli] to 2,4-dichlorophenoxyacetic acid or 2,4,5-trichlorophenoxyacetic acid results in the semisynchronous production of somatic embryos starting about 4 days after transfer to a non-auxin-containing medium. The timing of cell division induction in the petiole explants was found to vary depending on the petiole tissue type. Cells adjacent to the vascular bundles divide first at about 48 h after exposure to auxin, closely followed by those of the inner parenchyma, whereas most of the cells of the subepidermal and epidermal layers start to divide later, between 72 and 120 h. Two different sources of callus were also evident. Cells adjacent to the vascular bundles and the inner parenchyma cells were the primary source of callus when a short, 2-day (non-embryo-producing) exposure to auxin was used. In contrast, the subepidermal and epidermal cells were the primary source of callus tissue when a longer, 10-day (embryo producing) exposure was used. It is concluded that the source of somatic embryos is primarily the daughter cells of the subepidermal or epidermal tissue or both.     

Cross-reactivity between storage and dust mites and between mites and shrimp

Many patients have sensitivities to multiple species of storage and house dust mites. It is not clear if this is because patients have multiple sensitivities to species-specific mite allergens or if these mites share many cross-reacting allergens. Our objective was to further define the cross-allergenicity between several species of storage and house dust mites using crossed-immunoelectrophoresis (CIE), crossed-radioimmunoelectrophoresis (CRIE), immunoblotting, and ELISA. CIE and CRIE reactions revealed that storage mites shared two cross-antigenic molecules and one of these bound IgE in a serum pool from mite allergic patients. Antibody in anti-sera built to each species of mite recognized many SDS–PAGE resolved proteins of other mite species and this suggested the potential for other cross-reactive allergens. Among patient sera, IgE bound to many different proteins but few had IgE that bound to a protein with common molecular weights across the mite species and this suggested mostly species-specific allergens. Antiserum built to each mite species precipitated one protein in shrimp extracts that bound anti-Der p 10 (tropomyosin) and IgE in the serum pool. Anti-Der p 10 showed strong binding to shrimp tropomyosin but very little to any of the mite proteins. ELISA showed the mite extracts contained very little tropomyosin. The storage and dust mites investigated contain mostly species-specific allergens and very small amounts of the pan-allergen tropomyosin compared to shrimp and snail.