Ornamentation of thecal plates in Protoperidinium (Dinophyceae) as seen by scanning electron microscopy

The thecae of over thirty species of Protoperidinium from aroundthe British Isles have been examined by SEM and the varioustypes of ornamentation described. The simplest type consistsof smooth plates in which the trichocyst pores have an evenor irregularly thickened rim. In one species the pores are situatedon mounds. In another variation the plates are covered withminute pimples. Many species have plates which are thickenedby a surface reticulum and this may be made up of lines of pimples,of irregularly arranged ridges or bars which protrude to formsmall spines. In one species the ridges always surround trichocystpores and in another there are prominent spines which bear noobvious relationship with the pores. It is concluded that thecalornamentation can provide useful taxonomic characters withinthis genus.     

Sexual and Asexual Processes in Protoperidinium steidingerae Balech (Dinophyceae), with Observations on Life-History Stages of Protoperidinium depressum (Bailey) Balech (Dinophyceae)

ABSTRACT. A suite of morphological, histological, and molecular techniques was used to reveal for the first time division, sexuality, mandatory dormancy period of hypnozygotes, and identity of life-history stages of any Protoperidinium spp. In both Protoperidinium steidingerae and Protoperidinium depressum , asexual division occurred by eleutheroschisis within a temporary cyst, yielding two daughter cells. Daughter cells were initially round and one-half to two-thirds the size of parent cells then rapidly increased in size, forming horns before separating. Gamete production and fusion was constitutive in clonal and non-clonal cultures, indicating that both species may be homothallic. Gametes were isogamous, approximately half the size and lacking the pink pigmentation of the vegetative cells, and were never observed to feed. Gamete fusion resulted in a planozygote with two longitudinal flagella. Planozygotes of P. steidingerae formed hypnozygotes. The fate of planozygotes of P. depressum is unknown. Hypnozygotes of P. steidingerae had a mandatory dormancy period of ca. 70 days. Germination resulted in planomeiocytes with two longitudinal flagella. Nuclear cyclosis occurred in the planozygotes of P. depressum , but in the planomeiocytes of P. steidingerae . The plate tabulation and gross morphology of gametes of P. steidingerae and P. depressum differed markedly from those of vegetative cells. Thus, misidentification of morphologically distinct life-history stages and incomplete examination of thecal plate morphology in field specimens has likely led to taxonomic confusion of Protoperidinium spp. in previous studies.     

Healing of microvascular arterial anastomoses, as seen on corrosion casts by scanning electron microscopy.

We made 44 microsurgical anastomoses in the femoral artery in rats. The healing was studied from one day to 4 months, by making an acrylic cast of the lumen of the femoral artery and the surrounding vessels, and examining the cast by scanning electron microscopy. The method proved to be easy and relatively quick, and it limited the possibility of artifacts. The high depth of focus permitted simultaneous study of the main lumen and of the perivascular structures after microvascular anastomoses.     

Mammalian fertilization as seen with the scanning electron microscope

For several years we have been looking at mammalian gametes and their interactions with the scanning electron microscope (SEM). Examining the images produced by the SEM has given us a three-dimensional view of sperm, eggs, and egg investments. We are particularly impressed with the structural variation among gametes of different mammalian species. In this short report we examine the structure of mammalian spermatozoa, eggs, zonae pellucidae, and cumuli. Our observations and those of others have led us to believe that variation in gamete structure and function may have evolved as a mechanism for reproductive isolation of mammalian species.     

Architecture of chitin gel as examined by scanning electron microscopy

An architecture of the solid phase of chitin gel was examined by scanning electron microscopy. The ultrastructure of the xerogel was microporous with parallel channels surrounded with membranous walls. The pore shapes at cross section were polyhedral with three walls at the junctions. The pore was 30–50 μm in diameter and 80–300 μm in length, and the thickness of the walls was less than 1.5 μm. The gel is considered to be a polyphasic gel, consisting of small droplets of water held up in these pores.     

Structure of trichomonads as revealed by scanning electron microscopy.

A scanning electron microscope (SEM) study of Hypotrichomonas acosta (Moskowitz), Trichomonas vaginalis Donné, Pentatrichomonas hominis (Davaine), and Tritrichomonas foetus (Riedmüller) provided new information about the structure of the periflagellar canal; emergence of the flagella from the cell body; structure of the undulating membrane; and position, shape, and size of the pelta. Of special interest were the spatial relationships of the attached part of the recurrent flagellum and the accessory filament in Hypotrichomonas and in the members of Trichomonadinae, i.e. Trichomonas and Pentatrichomonas.     

Zona reaction in porcine oocytes fertilized in vivo and in vitro as seen with scanning electron microscopy

Morphological changes in zona pellucidae (ZP) isolated from in vitro-matured (IVM) and ovulated porcine oocytes were compared before or after fertilization in vitro and in vivo, respectively, by using scanning electron microscopy (SEM). The ZP of some ovulated or IVM oocytes and in vivo- or in vitro-fertilized (IVF) zygotes were equally split into two halves while immersed in an enzyme-inhibitor solution, using a surgical blade. After washing, intact and ZP halves were fixed in 1% glutaraldehyde solution in 0.1 M cacodylate buffer, processed, and examined using SEM. The outer surface of ZP in ovulated oocytes had a mesh-like structure. The outer morphology in IVM oocytes was more smooth although the mesh-like structure was still visible at high magnification. In in vivo zygotes and IVM-IVF zygotes, this lysed, mesh-like structure was more obvious. The inner surface of ZP had some small depressions (orifices). The mean number of orifices per 100 micrometer(2) of ZP surface was larger in IVM oocytes as compared to ovulated ones. The number of orifices per 100 micrometer(2) decreased in IVM-IVF zygotes as compared to IVM oocytes; whereas, in vivo zygotes did not differ from ovulated oocytes. The mean diameter of intact ZP as well as their mean thickness was greater in ovulated oocytes than IVM oocytes. The mean thickness of the ZP was larger in ovulated oocytes than IVM ones. The ZP thickness was larger in zygotes than in in vivo oocytes, whereas that of IVM-IVF zygotes did not differ from that of IVM oocytes. These results indicate that the morphology of ZP and the ZP reaction at sperm penetration appears to be much different between IVM oocytes and ovulated ones.     

The angioarchitecture of estrous, pseudopregnant and pregnant rabbit ovary as seen by scanning electron microscopy of vascular corrosion casts

Ovarian angioarchitecture was studied by scanning electron microscopy of vascular corrosion casts in estrous, pseudopregnant (stimulated with human chorionic gonadotropin) and pregnant rabbits. In all samples, the proper ovarian branch of the ovarian artery (ramus ovaricus) entered the ovarian hilus near the caudal pole of the organ and ran parallel to the major axis of the hilus. The extraovarian venous drainage was formed by several vessels emptying into a distal large vein. The ramus ovaricus exhibited various degrees of coiling and branched in the medulla. The coiling of the ramus ovaricus and its ramifications were maintained in all samples. A venous meshwork and/or flat vein branches closely enveloped the arterial coils found in the hilus and outer medulla. At this level numerous arteriovenous contacts were demonstrated in all samples. The coiled arteries, prior to entering the ovarian cortex, supplied several small peripheral follicles which were drained by the hilar veins. In the cortex the coiled arteries branched in numerous thin, straight or slightly undulated arterioles which supplied developing estrous follicles and pseudopregnant corpora lutea. The arterioles supplying the pregnant corpora lutea were long, large and tightly spiraled. The venous drainage followed the modifications of the arterial supply. These data demonstrate that ovarian cycle and pregnancy induced significant changes in the cortical vessels, which adapted their structure to the temporary functional needs of the recruited follicles or corpora lutea. Hilar and medullary vessels have permanent structures that may represent morphological devices for (a) a continuous control of the blood flow (spiral arteries) and (b) a local recirculation of endocrine products (arteriovenous contacts) comparable to the ”countercurrent mechanism” previously shown to operate in ovaries of other species, but not yet found in rabbits. Received: 19 June 1996 / Accepted: 7 October 1996     

Myofibrillogenesis in vitro as seen with the scanning electron microscope

The present study describes an experimental approach whereby myofibrillogenesis and the three-dimensional arrangement of myofibrils present within cultured skeletal muscle cells can be examined using the scanning electron microscope. This procedure uses cells that have been cultured on gold-coated coverslips, and treated with Triton X-100 to extract the cell membrane and the soluble cytoplasm. Subsequent electroconductive staining by treatment with thiocarbohydrazide and osmium allows the myofibrils to be visualized. The images of myofibrils in various states of development observed by this method generally accords to those previously reported by transmission electron microscopy. Cell elongation and adhesion to the substrate causes mechanical stress from different directions which meet at branchings of the cultured myotubes. Many myofibrils are observed to run in the direction of the inferred stress lines.     

Ultrastructure of Proechinophthirus zumpti (Anoplura,Echinophthiriidae) by scanning electron microscopy

The ultrastructure of Proechinophthirus zumpti Werneck, 1955, mainly the external chorionic features of the egg, is described through electronic microscopy techniques. This species was first cited in Argentina, infesting Arctocephalus australis (Zimmermann, 1873). The morphological adaptations of adults and nymphs are described in both species of Proechinophthirus parasitic on Otariidae: P. fluctus (Ferris, 1916) and P. zumpti.     

Laser microdissection of fungal septa as visualised by scanning electron microscopy

Laser microdissection has been proven a successful technique to isolate single cells or groups of cells from animal and plant tissue. Here, we demonstrate that laser microdissection is suitable to isolate subcellular parts of fungal hyphae. Dolipore septa of Rhizoctonia solani containing septal pore caps were cut by laser microdissection from sections of mycelium and collected by laser pressure catapulting. Subsequently, microdissected septa were visualised using a wheat germ agglutinin labelling of cell walls, septa and septal pore caps and scanning electron microscopy. The use of laser microdissection on fungal cells opens new ways to study subcellular fungal structures and the biochemical composition of hyphal cells.     

The maxillae of Drosophila melanogaster as revealed by scanning electron microscopy

The scanning electron micrographs show the external morphology of the maxillae of Drosophila melanogaster. Specifically, they illustrate the patterning of the different types of chemo-receptive sensilla on the maxillary palpi making possible a clearer understanding of the structure of the tiny maxillary lobes. It appears that the maxillary lobes act as “cleaning brushes” during the feeding process.     

Molecular phylogenetic study of the heterotrophic dinoflagellate genus Protoperidinium (Dinophyceae) inferred from small subunit rRNA gene sequences

In the present study, we investigated the intrageneric and intergeneric phylogenetic relationships of the heterotrophic marine dinoflagellate genus Protoperidinium. Using single‐cell polymerase chain reaction methods, we determined small subunit ribosomal RNA gene sequences for 10 Protoperidinium species belonging to four sections and two subgenera. Phylogenetic trees were constructed using maximum parsimony, neighbor joining and maximum likelihood methods. We found intraspecific variability of small subunit rDNA sequences in Protoperidinium conicum (Gran) Balech, Protoperidinium crassipes (Kofoid) Balech and Protoperidinium denticulatum (Gran et Braarud) Balech, but not in other species. The small subunit rDNA phylogeny revealed that the genus is monophyletic, but its phylogenetic position within the Dinophyceae could not be determined because of ambiguous basal topologies. Within the genus Protoperidinium, species of the subgenus Archaeperidinium with two anterior intercalary plates (2a) were shown to be monophyletic, but species of the subgenus Protoperidinium with three anterior intercalary plates (3a) were resolved as paraphyletic. The sections Avellana, Divergentia and Protoperidinium were shown to be monophyletic, while the section Conica was paraphyletic. Based on the trees obtained in the present study, most of the traditionally defined sections are supported by molecular phylogeny. It was also indicated that the section Avellana evolved from one of the Conica‐type dinoflagellates.     

Surface structures and sense organs of the cypris larva of Balanus balanoides as seen by scanning and transmission electron microscopy.

Various features of the settlement stage larva (cyprid) of the barnacle, Balanus balanoides (L.), were studied using scanning and transmission electron microscopy. The cuticle of the valves is pitted and in section has a characteristic ultrastructure. Small sensory setae protrode from the surface of this cuticle and are probably mechanoreceptors able to sense water movement around the larva. Each of the pair of caudla appendages which protrude from between the larval valves posteriorly, is made up of several sensory setae. These appendages are able to sense settlement surface topography. Certain other features of the larva are alos described and their roles discussed; such features include the frontal filaments, antennules and thoracic limbs.