Gamete interaction in the white sturgeon Acipenser transmontanus: a morphological and physiological review

Synopsis Sturgeon gametes differ from those of most fish in that the sperm possess acrosomes that undergo exocytosis and filament formation while the eggs possess numerous micropyles. Acipenser transmontanus eggs are encased by multilayered envelopes that consist of outer adhesive jelly coats and three structured layers interior to the jelly. The glycoprotein jelly layer only becomes adhesive upon exposure to freshwater. The layer interior to the jelly, layer 3, is the other carbohydrate-containing component of the egg envelope. This layer consists of a water-insoluble glycoprotein that, upon freshwater exposure, is hydrolyzed by a trypsinlike protease to yield a water-soluble, lower molecular weight carbohydrate-containing component. This component can be identified in the surrounding medium when unfertilized eggs are incubated in freshwater. This egg water component elicits acrosome reactions only in homologous sperm. The A. transmontanus sperm acrosome reaction is a Ca⁺⁺ and/or Mg⁺⁺ dependent event that includes the formation of a 10 μ long fertilization filament. A. transmontanus fertilization can occur at low sperm per egg ratios; however, crossfertilization of A. transmontanus eggs with lake sturgeon, A. fluvescens, sperm results in a very low number of fertilized eggs, even at high sperm per egg ratios. The morphological, physiological, and biochemical phenomenon reviewed in this paper are related to the environment in which they occur. Also, the possible role of the acrosome and the presence of numerous micropyles are discussed.     

Previtellogenic and vitellogenic oocytes in ovarian follicles of cultured siberian sturgeon Acipenser baerii (Chondrostei,Acipenseriformes)

Previtellogenic and vitellogenic oocytes in ovarian follicles from cultured Siberian sturgeon Acipenser baerii were examined. In previtellogenic oocytes, granular and homogeneous zones in the cytoplasm (the ooplasm) are distinguished. Material of nuclear origin, rough endoplasmic reticulum, Golgi complexes, complexes of mitochondria with cement and round bodies are numerous in the granular ooplasm. In vitellogenic oocytes, the ooplasm comprises three zones: perinuclear area, endoplasm and periplasm. The endoplasm contains yolk platelets, lipid droplets, and aggregations of mitochondria and granules immersed in amorphous material. In the nucleoplasm, lampbrush chromosomes, nucleoli, and two types of nuclear bodies are present. The first type of nuclear bodies is initially composed of fibrillar threads only. Their ultrastructure subsequently changes and they contain threads and medium electron dense material. The second type of nuclear bodies is only composed of electron dense particles. All nuclear bodies impregnate with silver, stain with propidium iodide, and are DAPI‐negative. Their possible role is discussed. All oocytes are surrounded by follicular cells and a basal lamina which is covered by thecal cells. Egg envelopes are not present in previtellogenic oocytes. In vitellogenic oocytes, the plasma membrane (the oolemma) is covered by three envelopes: vitelline envelope, chorion, and extrachorion. Vitelline envelope comprises four sublayers: filamentous layer, trabecular layer 2 (t2), homogeneous layer, and trabecular layer 1 (t1). In the chorion, porous layer 1 and porous layer 2 are distinguished in most voluminous examined oocytes. Three micropylar cells that are necessary for the formation of micropyles are present between follicular cells at the animal hemisphere. J. Morphol. 278:50–61, 2017. ©© 2016 Wiley Periodicals,Inc.     

Electron Microscopic Observations on the Cortical Reaction of the Brittle-Star Amphipholis kochii Lütken, with Special Reference to its Vitelline Coat Modification as Revealed by the Surface Replica Method

This paper describes the fine structural changes of the egg of the brittle-star Amphipholis kochii Lütken during the cortical reaction. The vitelline coat is 20 nm thick, when Ruthenium Red stain is used, and consists of a dense network of fibers. The cortical granules are large, 1.5–2.0 μm in diameter, and exist in several layers in the egg cortex, unlike the monolayer arrangement found in many other animals. The contents of the cortical granules are clearly distinguished into two components: peripheral fibrous (PF) material and central fibrous (CF) material that consists of two components differing in electron density. The PF material is densely stained by periodic acid-chromic acid-silver methenamine stain, while the CF material is stained little if at all by this technique. The vitelline coat and some PF materials form the fertilization membrane, which is about 40 nm thick and consists of three layers; the outer and the inner layer of the fertilization membrane each have a trilaminated structure. The vitelline coat substances are probably located in the upper part of the fertilization membrane. The hyaline layer, 7–8 μm thick, consists mainly of CF materials. These observations on the morphology of the ophiuroid egg are discussed in comparison with those on other echinoderms, especially echinoids and asteroids.     

Ultrastructure of the chorion and its micropyle apparatus in the mature Fundulus heteroclitus (Walbaum) ovum

The ovum 'membranes' and the micropyle apparatus of mature, extruded ova of Fundulus heteroclitus were examined by scanning electron microscopy. The ovum is covered by a thin jelly coat comprised of a dense mat of 0.3-0.5 urn fibrils, except for a 50–100 um fibril-free zone surrounding the micropyle apparatus. The micropyle apparatus consists of a 50–100 um diameter funnel-shaped vestibule, at the bottom of which is a circular aperture 4–5 μ in diameter leading to the micropyle canal which traverses the entire chorion layer. The inner micropyle aperture, 2–3 um diameter, apposes the inner ovoplasm mass.
The chorion is the major protective coating of the ovum. It consists of a thin (0.4 μm) outer zone, a thicker (9–12 μ), lamellated inner zone with 4–10 lamellae, and sometimes an innermost crystalline zone, varying in thickness from 1–13 μm. The extreme variability in the structure of the lamellated and crystalline zones of the chorion suggests that generalizations concerning ovum membranes can be misleading.     

Myxosoma microspora n. sp. (Myxosporidia: Protozoa) parasitic in the gills of Mugil cephalus L.

A new histozoic species of myxosporidian, Myxosoma microspora n.sp., infecting the gill filaments of Mugil cephalus is described. Cysts measuring 0.5–1.0 mm in diameter were found attached to the gill filaments. Spherical or slightly oval, spores 4.8–5.2 μm in diameter, were present and possessed a thin outer mucous envelope which appeared as small conical protuberances at the ends of the equatorial axis. Polar capsules were pyriform in shape, equal in size and measured 1.6–2.0 × 1.0–1.2 μm; the polar filaments were 22–28 μm in length. There was a bean-shaped sporoplasm measuring 3.5 × 1.5 μm. No iodine vacuole was detected when the parasite was stained with Lugol's iodine.     

The fine structure of the eggshell of the laid egg of Locusta migratoria migratorioides

Ultrastructural analysis disclosed a regional diversification of the eggshell of the Locusta migratoria m. (L.m.m.) egg. This diversification is discussed taking into account the data about water permeability of the L.m.m. egg. The presence of several kinds of openings in the exochorion implies that in addition to the micropyles, the L.m.m. egg is provided with a respiratory system, especially in its anterior pole. The respiratory system of insect eggs is briefly reviewed. The presence of a wax layer in the L.m.m. egg is also discussed.     

Development of the pollen grain wall in Canna

During the tetrad period spinules form on the Canna L. plasma membrane at intervals of 1–2 μm on a microspore surface of ca. 100 μm². The isolated spinules represent all that there is of a primexine-like nature. Immediately following loss of the callosic tetrad envelope a channeled, oncus-like zone forms on the plasma membrane over the entire microspore surface, elevating the spinules. The oncus-like zone becomes ca. 4 μm thick by microspore mitosis. Intine introduction during the pollen grain period coincides with substantial thinning of the oncus-like zone and pollen grain enlargement. In the final phases of maturation grains increase further in diameter and become packed with starch and lipoidal material. The oncus-like zone more than doubles in height necessitating a migration of the boundary of the oncus-like zone and intine. At pollen grain maturity the thin (ca. 200 nm) surface layer of the oncus-like zone appears to be compacted or filled in.     

Eimeria procyonis sp. n., an Isospora sp., and a Redescription of E. nuttalli Yakimoff and Matikaschwili, 1932 (Protozoa: Eimeriidae) from the American Raccoon (Procyon lotor)

SYNOPSIS. Thirty-two of 48 raccoons examined were infected with a previously undescribed species of Eimeria which is herein named E. procyonis. Of the 32 infected animals, 10 also harbored E. nuttalli and 1 had Isospora sp. oocysts.
The ellipsoid to ovoid oocysts of E. procyonis measured 23.4 × 18.0 (16–29 × 13–24) μm; its sporocysts measured 12.1 × 9.3 (11.5–15 × 7–10) μm, each containing a slightly flattened substiedal body. The sporocyst residuum consisted of numerous scattered granules each ∼1 μm in diameter. The oocyst wall was double-layered. The outer layer appeared rough and pitted, measuring 1.5 μm, except at the micropyle where it was 1 μm thick.
The oocysts of the Isospora sp. measured 16.8 × 13.7 (16–18.5 × 12.5–15.5) μm. The wall consisted of a single layer ∼0.5 μm thick. The sporocysts measured 11.2 × 9.1 (9.5–11.5 × 8–10) μm, and each contained 4 elongate sporozoites. The oocysts of E. nuttalli measured 17.5 × 13.6 (12-21 × 11-15) μm, with a smooth single-layered wall approximately 0.7 μm thick. The sporocysts measured 12.2 × 7.1 (9-13 × 5.5–11) μm. Each sporocyst had a thin, dark, Stieda body and the sporocyst residuum consisted of many fine granules.     

Light and Electron Microscopy of the Spore of Myxobolus heckelii n. sp. (Myxozoa), Parasite from the Brazilian Fish Centromochlus heckelii (Teleostei, Auchenipteridae)

ABSTRACT. A myxosporean parasitizing the gill filaments of the freshwater teleost fish Centromochlus heckelii collected in the Tocantins River (Lower Amazonian Region, Brazil) is described using light and electron microscopy. This parasite produces spherical to ellipsoidal cyst-like plasmodia up to 250 μm in diameter, with a thick wall strengthened by several stratified juxtaposed crossed collagen layers, whose thickness varies according to the number of the layers. Several compressed fibroblasts are observed among the collagen fibrils. Deposits of spherical dense material are scattered at the internal periphery of the cysts. Plasmodia and different developmental stages, including immature and mature spores, filled the central region of the cysts. The spore body is ellipsoidal in valvar view and biconvex in sutural view. It is formed by two equal-sized and symmetric valves measuring 12.7 μm long (12.2–13.1) ( n =50), 6.6 μm wide (6.3–6.9) ( n =25), and 4.0 μm (3.7–4.4) ( n =20) thick. A thin layer formed by fine and anastomosed microfibrils is observed at the spore surface. Two equal, elongated pyriform polar capsules measure 2.9 μm (2.7–3.3) × 1.7 μm (1.4–2.0) ( n =25), each containing four or five oblique polar filament coils. The binucleated sporoplasm contains numerous spherical sporoplasmosomes, glycogen particles, and a large vacuole with fine granular matrix. Based on the morphological and ultrastructural differences and specificity of the host, we describe this isolate as a new myxosporidian, Myxobolus heckelii n. sp. (Myxozoa, Myxosporea).     

Structural Changes of Yolk Platelets and Related Organelles during Development of the Newt Embryo

Structural changes in yolk platelets and related organelles in the cytoplasm of the presumptive ectodermal region up to the stage of gastrulation were studied by light and electron microscopies using full-grown oocytes, mature eggs descending the oviduct and embryos of the newt, Cynops pyrrhogaster . Yolk platelets with a superficial layer are first observed in mature eggs descending the oviduct. During the cleavage and early morula stages, the superficial layer increases in thickness and the main bodies become more slender. The superficial layer decreases in thickness in the blastula stage, and many yolk platelets lose this layer in the gastrula stage.
The amount of rough-surfaced endoplasmic reticulum (r-ER) increases rapidly in the morula stage, while Golgi complexes gradually increase in number between the cleavage and gastrula stages. In the cleavage and early morula stages, most of the r-ER is closely adherent to yolk platelets and is associated with several mitochondria. Two types of free vesicles, large (0.5–4.0 μm diameter) and small (0.15–0.3 μm diameter), were seen in abundance from the early morula stage to the early gastrula stages.
Changes in the structure of yolk platelets are discussed in relation to changes in other cytoplsmic organelles.     

Formation and structure of egg envelopes in Russian sturgeon Acipenser gueldenstaedtii (Acipenseriformes: Acipenseridae)

The covering of the eggs in Russian sturgeon Acipenser gueldenstaedtii consists of three envelopes (the vitelline envelope, chorion and extrachorion) and is equipped with multiple micropyles. The most proximal to the oocyte is the vitelline envelope that consists of four layers of filamentous and trabecular material. The structural components of this envelope are synthesized by the oocyte (primary envelope). The chorion encloses the vitelline envelope. The extrachorion covers the external surface of the egg. Examination of the arrangement of layers that comprise the egg envelopes together with the ultrastructure of follicular cells revealed that the chorion and extrachorion are secondary envelopes. They are secreted by follicular cells and are built of homogeneous material. During formation of egg envelopes, the follicular cells gradually diversify into three morphologically different populations: 1) cells covering the animal oocyte region (cuboid), (2) main body cells (cylindrical) and (3) micropylar cells. The apical surfaces of follicular cells from the first two populations form processes that remain connected with the oocyte plasma membrane by means of gap junctions. Micropylar cells are located at the animal region of the oocyte. Their apical parts bear projections that form a barrier to the deposition of materials for egg envelopes, resulting in the formation of the micropylar canal.     

The development of the hexagonally structured egg envelope of the C-O sole (Pleuronichthys coenosus)

The surface of a mature, pelagic C-O sole egg is composed of polygonal chambers having four to eight sides, most of which are hexagonally shaped. This honeycomb pattern initially appears on primary oocytes as a thin layer of compact, electron-dense material. Discrete thickenings begin to develop on the envelope of perinuclear stage oocytes. The thickenings lengthen and thin to form the hexagonal walls of the envelope in oocytes undergoing yolk vesicle formation. The walls of each hexagonal chamber occur in an area corresponding to the lateral margins of the adjacent follicle cell, suggesting that the hexagonal walls are produced by the follicle cells. The hexagonal layer is nearly complete at the beginning of vitellogenesis, and as vitellogenesis continues, a striated envelope layer composed of fibrillar lamellae develops between the oocyte and the hexagonal layer. The striated layer appears to be secreted by the oocyte. After vitellogenesis is completed, oocytes are ovulated and double in size during a period of maturation. Concurrently, the striated primary envelope stretches and thins into eight to nine horizontal lamellae. On the mature egg surface, the polygonal chambers are about 24–31 μm in diameter. Within each chamber there is a subpattern of polygonal areas; each polygon is 1.5–2.0 μm in diameter, and circumscribes a pore canal opening. This exceptional envelope may furnish the egg with some degree of protection, resiliency, and buoyancy, but its specific functions are not known.     

The Structure of the Oocyst and the Excystation Process of Isospora marquardti sp. n. from the Colorado pika, Ochotona princeps

SYNOPSIS. Oocysts of Isospora marquardti sp. n. from the Colorado pika, Ochotona princeps , have spheroid oocysts, 30.5 (23–36) μm in diameter, and ovoid sporocysts, measuring 19.3 × 12.0 (17–22 × 10–14) μm. A polar body, 2 × 4 μm, a spheroid sporocyst residuum (8.3 μm in diameter), a Stieda body, and a distinct substiedal body (3 × 3 μm) are present. A micropyle and an oocyst residuum are absent. Excysted sporozoites, averaging 3.0 × 18.5 (2–4 × 15–20) μm, contain 2 refractile globules, 1 on each side of the nucleus with a prominent nucleolus.
The sporozoite excystation process using a trypsin-sodium taurocholate fluid is described.     

Induced spawning of grey mullet Mugil cephalus L. with fractionated salmon pituitary extract

The efficacy of fractionated salmon pituitary gonadotropin as a spawning agent in Mugil cephalus L. was tested. Natural spawning was induced in all females with a total dose of 11.9–20.9 μg/g body wt. Spawning dose varied inversely with initial mean egg diameters of recipient females. A 'critical' mean egg diameter of 650–700 μ was observed to precede the hormone dose that induced spawning. A 'priming' effect was observed following the initial injection and is discussed. The 'latency period' was determined to be 10–15 h; fecundity was estimated at 648 eggs/g body wt. Courtship, spawning and fertilization occurred naturally with uninjected males.     

Comparative morphology of the egg of the castniid palm borer,Paysandisia archon (Burmeister, 1880) (Lepidoptera: Castniidae)

Paysandisia archon (Burmeister, 1880) is an attractive neotropical castniid moth whose presence in Europe was recently reported. Its larvae are endophagous, feeding inside the trunks and branches of several species of palm trees (Arecaceae). The present paper deals with the morphology and biometry of the egg of this moth, comparing them with those of other castniid species.

The egg is a typical castniid egg, fusiform, upright sensu Döring, light cream or creamy pink when freshly laid, 4.69 ± 0.37 mm long and 1.56 ± 0.11 mm wide. Larvae emerge by gently splitting the chorion along one of the longitudinal ridges, on the half closer to the micropyle. SEM, TEM and LSCM photographs showing ultrastructural details of the egg are shown for the first time. The micropylar rosette (c. 54 μm in diameter) has generally 14–17 cells; in its centre lies the micropylar pit (c. 6 μm in diameter) which bears 12–16 micropylar canal openings (= micropyles) around its periphery. The pathways followed by those canals through the chorion have been figured. Eggs sampled in the wild (so laid by several females) were found to have a slightly variable number of ridges: most bore seven ridges (68.87%), although a significant portion (30.46%) bore eight and 1 egg (0.67%) bore only six; this against the currently accepted rule of five‐ridged eggs for Castniini (i.e. Neotropical castniids) to which Paysandisia archon belongs. It has also been found that the same female specimen has the capability of producing six‐, seven‐ or eight‐ridged eggs. Five types of egg irregularities affecting the longitudinal ridges are also figured and described. Transverse striae on the egg of P. archon are about 122. Aeropyles (c. 4 μm in diameter) occur on the ridges, at the intersections between the latter and two contiguous (left and right of the ridge) transverse striae, amounting to c. 854 on a seven‐ridged egg and to c. 976 on an eight‐ridged egg. Occasionally minute aeropyles ('microaeropyles') (c. 1.96–3.13 μm in diameter) also occur on transverse striae located close to both egg poles.

The chorion of P. archon shows the typical ditrysian fine structure with very thin basal layer (C‐1), 0.3–0.2 μm thick, gas‐filled trabecular layer (C‐2), c. 0.9 μm thick, and lamellar layer (C‐3), its thickness varying between 18.5 and 13 μm due to the bumpy external surface of the chorion. Aeropylar canals, that penetrate layer C‐3, connect the air‐containing inner chorionic meshwork (the trabecular layer C‐2) with the surrounding air; their outer part forms a big bulbous cavity (which opens to the outside through the small opening seen in external SEM images) and, underneath, a narrow canal follows, leading into the trabecular layer (C‐2).     

Filtering mechanism of the honey bee proventriculus

ABSTRACT. The fine structure and function of a honey bee's ( Apis mellifera Linn.) proventriculus were studied by scanning electron microscopy and video-recording. Our observations revealed that the proventriculus is used to engulf pollen and other particles which contaminate the nectar carried into the crop. The four lips are closed and opened, pulled backwards and straightened by the external circular muscles and internal longitudinal muscles. Combs of filiform-hairs (70 μm in length) located on the margins of the lips 'catch' and filter particles from the fluid. By repeated filtering, opening and closing actions of the hairs and lips, particles are filtered and collected in pouches between the ventricular folds to form boluses and are eventually passed into the midgut. In the present experiment, particle sizes ranging from 0.5 to 100 μm in diameter, including dandelion pollen ( Taraxacum officinale Web.), Torula yeast ( Candida utilis Lodder et Kreger-Van Rij), bee disease spores of Nosema apis Zander and Bacillus larvae White, and man-made particles can be filtered by the hairs. Small particles (0.23 μm in diameter) filter through the hair and return back to the fluid. Large particles (100–200 μm in diameter) are caught between the stylets of the mouthparts and are not ingested. These observations suggest that the particle size plays an important role in determining what can be taken by the mouthparts and the proventriculus and what can later be utilized as a food source by the bee. The role of the proventriculus in disease transmission is also discussed.     

Egg structure of Zorotypus caudelli Karny (Insecta, Zoraptera, Zorotypidae)

The structural features of eggs of Zorotypus caudelli Karny are described in detail. The egg is elliptic with long and short diameters of 0.6 and 0.3 mm respectively, and creamy white. The egg shows a honeycomb pattern on its surface, without any specialized structures for hatching such as an operculum or a hatching line. The fringe formed by a fibrillar substance secreted after the completion of the chorion encircles the lateral surface. The egg layer is composed of an exochorion, an endochorion, and a vitelline envelope. The exochorion and endochorion are electron-dense and homogeneous in structure. The exochorion shows a perforation of numerous branching aeropyles. The exo- and endochorion are connected by numerous small columnar structures derived from the latter. The vitelline envelope is very thin and more electron-dense than the chorion. A pair of micropyles is present at the equator on the dorsal side of the egg. Originating at the micropyle, the micropylar canal runs through the chorion obliquely. The structural features of the eggs of Zoraptera were compared with those of other polyneopteran and paraneopteran orders.     

ULTRASTRUCTURAL OBSERVATIONS ON THE DISCHARGE OF TWO KINDS OF GRANULES IN THE FERTILIZED EGGS OF CYPRINUS CARPIO AND CARASSIUS AURATUS

This paper describes mainly ultrastructural evidence on the discharge of two kinds of granules, CA and CB, as well as the cortical alveoli in an egg of the common carp and the goldfish. The cortical alveoli (about 2–28 μ in diameter) and CA-granules (about 0.4–2 μ in diameter) are located in the cortical cytoplasm of the mature egg, and the latter is distinguishable in size and texture from the former which contains an eccentric core at the very least as a basal internal structure. The CB-granules (about 100–950 mμ in diameter) appear in small or large clusters in the cortical cytoplasm after fertilization, being formed in connection with constriction or pinching-off of dilated tubular elements. After fertilization the cortical alveoli, CA- and CB-granules are discharged at different times. The new plasmalemma of a fertilized egg appears to be a joint production of the original plasmalemma and the limiting membranes of the cortical alveoli, CA- and CB-granules.     

Eimeria crotalviridis sp. n. from Prairie Rattlesnakes, Crotalus viridis viridis, in New Mexico with Data on Excystation of Sporozoites and Ultrastructure of the Oocyst Wall

SYNOPSIS Oocysts of Eimeria crotalviridis sp. n. are described from prairie rattlesnakes, Crotalus viridis viridis in New Mexico on the basis of light and electron microscopy and in vitro excystation of sporozoites. Sporulated oocysts of E. crotalviridis are elliptical, 26.4 × 22.3 (23–29 × 20–24) μm with ovoid sporocysts 11.7 × 8.1 (11–13 × 7–9) μm. A micropyle, micropyle cap and polar bodies are absent, but oocyst and sporocyst residua and Stieda and substieda bodies are present. Excysted sporozoites are 12.4 × 2.8 (11–13 × 2–3) μm and have 1 large posterior refractile body and a nucleus with a prominent nucleolus. Ultrastructurally, the oocyst wall has 2 layers, a thick, electron-dense, highly sculptured outer layer composed of a fine granular matrix and a thin, granular, osmiophilic inner layer, separated from the outer layer by at least one unit membrane. These layers are 441 (353–510) and 21.6 (19–29) nm thick, respectively. Within 15 min after exposure to a trypsin-sodium taurocholate fluid, sporozoites of E. crotalviridis excysted from 5-month-old sporocysts.     

Time Sequence of Early Events in Fertilization in the Medaka Egg

The time sequence of early events in fertilization was examined in eggs of the medaka Oryzias latipes . The mean time after insemination required for sperm attachment to the egg surface through the micropyle depended on sperm concentrations. It was 3 ± 1 sec with a range from 1 to 6 sec after insemination when concentration of spermatozoa was high (about 2 × 10⁸/ml at 23°–25°C). The mean time from sperm attachment until cessation of its movement on the egg surface was 4 ± 1 sec with a range from 1 to 9 sec. Small cortical alveoli at the animal pole region within 15 μm of the sperm attachment point began to undergo exocytosis 9 ± 0.3 sec (range 5–16 sec) after sperm attachment. The velocity at which the exocytosis wave propagated increased from the earliest initiation point of exocytosis up to the 100 μm area, and became constant at about 12 μm/sec from 100 μm to 500 μm from the sperm attachment point. The present results suggest that at the time of fertilization in the fish egg, exocytosis of small cortical alveoli in the area about 15 μm away from the sperm attachment point occurs simultaneously.