Egg envelopes of Streptocephalus dichotomus Baird: A structural and histochemical study

The origin, ultrastructure and histochemical properties of the egg membranes in the South Indian anostracan, Streptocephalus dichotomus have been studied. The egg morphology and the ultrastructure of the tertiary membrane of this phyllopod crustacean have been examined both by scanning and transmission electron microscopy. Scanning electron microscopic observations on the egg surface reveal the characteristic ridges on the egg surface with pores. Similarly, the tertiary egg shell of S. dichotomus consists of two distinct layers, an outer cortex and an inner alveolar layer. There are specific differences in the structure and in the relationship of one layer to the other. The alveolar layer is characterised by large lipid droplets and an alveolar mesh. These two layers termed as tertiary layers are secreted by maternal shell glands. The outer tertiary egg layers are phenolically tanned, the precursor materials for tanning being derived from shell gland secretions.     

Subcortical Rotation and Specification of the Dorsoventral Axis in Newt Eggs

The specification of the dorsoventral axis in naturally polyspermic eggs of the Japanese newt, Cynops pyrrhogaster , was first examined by studies on the spatial relationship between the dorsal midline of the future body plan and the sperm entrance points (SEPs ¹ ). On local insemination, the dorsal blastopore lip was usually found to be formed opposite the SEPs, as in anuran monospermic eggs. Next the movements of the subcortical layer and the cortex were analyzed. "Subcortical rotation" was observed, similar to that of Xenopus laevis eggs with respect to its timing and extent, and its direction was shown to predict the embryonic axis of the eggs. Thus, the dorsoventral axis was concluded to be determined by essentially the same mechanism in the newt as in Xenopus .
Owing to their large size and long first cell cycle, newt eggs appear to be suitable material for study of subcortical rotation, but their behavior is unique in that subcortical rotation occurs in only the vegetal hemisphere so that the subcortical layer stretches in the future dorsal side. Studies on the movement of Nile blue spots suggested that the cytoplasm under the cortex in newt eggs consists of two layers.     

Fine structural investigation of the insemination response in Urechis caupo.

Electron microscopy of Urechis eggs revealed no changes in the egg cortex or investing layers until 4 min after insemination at 172C. From 4 min to about 30 min after insemination the surface coat gradually elevates, widening the perivitelline space. During this period, microvilli separate from the tightly woven layer of the surface coat, fibrogranular aggregates resembling surface coat material appear in the perivitelline space, and some cortical granules are extruded from the egg cortex into cytoplasmic processes. There is no statistically significant decrease in the number of cortical granules remaining in the egg surface during the first 95 min after insemination; many cortical granules persist in postgastrulae. Most of the cortical granules remain in fertilized eggs after removal of the surface coat with glucose-EGTA. We found no morphological correlates of the polyspermy block which is established within 1 min of insemination (Paul, 1975).     

Cholinergic structures in the cat auditory cortex (area AI)

Plexuses of cholinergic varicose fibers, differing in density in different layers of the neuropil, were found in area AI of the cat's auditory cortex by the histochemical reaction for acetylcholinesterase: Their density was maximal or average in layer I or deeper layers and minimal in layers II and III. Among cells in area AI those which are cholinergic are a few stellate neurons located in layers II–VI. Axons of some neurons terminate on neighboring cells, those of others (some neurons in layer VI) run into the subcortical layer of arcuate association fibers. Cholinergic terminals are located on the bodies and proximal areas of dendrites of neurons most of which do not contain acetylcholinesterase. Choliniceptive neurons of different sizes and shapes are found in all layers of this region of the auditory cortex.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. I. I. Mechnikov Odessa State University. Translated from Neirofiziologiya, Vol. 16, No. 1, pp. 75–81, January–February, 1984.     

Bipolar segregation of mitochondria,actin network,and surface in the Tubifex egg: Role of cortical polarity

The role of the cortex in ooplasmic segregation of the yolky eggs of Tubifex has been studied by epifluorescence microscopy. Living eggs labeled with rhodamine 123 and fine carbon particles placed on the surface showed that, following the second polar body formation, the egg surface cosegregates with subcortical mitochondria in a bipolar fashion, viz. toward the animal and vegetal poles in the animal and vegetal hemispheres, respectively. The egg surface of each pole moves spirally while the equatorial surface appears to remain stationary during this process. The rhodamine-phalloidin staining of whole eggs reveals that actin networks cosegregate with mitochondria. Isolated cortices which were stained with rhodamine-phalloidin demonstrated that cortical actin is organized bipolarly and that, during ooplasmic segregation, it undergoes reorganization directed toward both poles of the egg. The cortical polarity expressed as actin organization is not disrupted by centrifugal force sufficient to stratify the egg cytoplasm into five layers. The surface of a centrifuged egg moves according to the original cortical polarity. This surface movement is accompanied by the reorganization of cortical actin which appears to be identical to that in intact eggs. Other centrifugation experiments have demonstrated that the connection of the subcortical cytoplasm to the cortex is resistant to a centrifugal force of up to 650g. The nature of cortical polarity and its role in ooplasmic segregation are discussed in the light of the present results.     

内蒙古巴音满都呼晚白垩世棱齿龙蛋化石的发现

本文记述的恐龙蛋化石标本,采自内蒙古乌拉特后旗巴音满都呼上白垩统牙道黑达组中。蛋化石在蛋窝中排列的方式和蛋壳的显微结构特征与北美发现的含有可鉴定为棱齿龙胚胎骨骼的蛋化石基本相似,但还有一些差别,如蛋壳外表面不具纵向细纹,柱状层中鱼骨型纹饰不明显等。因此,应为棱齿龙科中另一新的属种代表。     

Intrinsic cholinergic components in cholinergic innervation of the cat auditory cortex (area AI)

Histochemical study of neuronally isolated area AI of the auditory cortex in cats by the reaction for acetylcholinesterase 3 days and 1, 2, and 3 weeks after undercutting showed that the cholinergic neuropil of this area is formed mainly by incoming fibers and to a lesser degree by processes from a few intrinsic cholinergic neurons. The intrinsic cholinergic neurons include, first, cholinergic long-axon association neurons responding to cortical isolation by retrograde changes and by hyperreaction to acetylcholinesterase (Cajal-Retzius cells of layer I and neurons of layer VI, whose axons run into the subcortical layer of association fibers), and, second, cholinergic short-axon association neurons of layers II–VI, preserving their normal cell structure and moderate acetylcholinesterase activity after isolation. Axon collaterals of similar cells terminate on neighboring neurons. Short-axon neurons are more numerous in the lower layers of the cortex, and exceed in number the long-axon association neurons. Choliniceptive neurons (pyramidal and stellate), on whose bodies and proximal dendrites are located terminals formed by axons of cholinergic association neurons, are found in the isolated cortex. Choliniceptive neurons are found more often in the lower layers of the cortex.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. I. I. Mechnikov State University, Odessa. Translated from Neirofiziologiya, Vol. 16, No. 1, pp. 81–87, January–February, 1984.     

FORMATION OF THE CORTICAL CONCAVITY AT FERTILIZATION IN THE SEA URCHIN EGG

During the initial stages of fertilization envelope elevation in eggs of Strongylocentrotus pur puratus and S. droebachiensis a large concavity of the egg cortex was observed in the light microscope. This concavity corresponded in shape and size with the elevating fertilization envelope. However, after the vitelline layers of eggs were disrupted and the eggs inseminated, the concavity failed to develop although the eggs were fertilized and developed normally. We propose that the concavity is formed owing to increased hydrostatic pressure within the perivitelline space. To further support this hypothesis we measured total egg protein secreted during fertilization, and found that 98% was retained within the perivitelline space. Furthermore, 80% of the total protein was contributed by the hyaline layer. Presumably, colloidal osmotic pressure and/or hydration of fertilization product, trapped beneath the fertilization envelope, is responsible for increased hydrostatic pressure within the perivitelline space, and therefore promotes not only fertilization envelope elevation, but the cortical concavity as well.     

ON PROPAGATION OF CORTICLA FACTOR AND CYTOPLASMIC FACTOR PARTICIPATING IN CLEAVAGE FURROW FORMATION OF THE NEWT'S EGG*

From Cynops pyrrhogaster eggs just after the start of the first cleavage, a fragment of cortical layer with a small entire cleavage furrow was cut out. In the fragment, the cortex had already acquired susceptibility to and the subcortical cytoplasm had already accquired inducibility for furrow formation. The fragment was transplanted to the animal hemisphere of uncleaved fertilized eggs or eggs immediately after the onset of the first cleavage, from which a portion of the host cortex was removed. Observation was made on division of the graft, and on propagation of the cortical susceptibility and the cytoplasmic inducibility of the graft onto the host egg. The transplant divided succesively on the host egg in many cases, but the furrow of the graft never advanced to the surface of the host egg. Neither the cortical factor nor the cytoplasmic factor was transmitted across the graft to the recipient egg.     

Dynamics of the actin microfilament system in the Tubifex egg during ooplasmic segregation

Following the second polar body formation (PBF), the Tubifex egg undergoes ooplasmic segregation consisting of two steps, i.e., centrifugal migration of membranous organelles forming a subcortical ooplasmic layer and then movements of these organelles along the egg surface. The present investigation was undertaken to examine the microfilament organization in eggs during these ooplasmic rearrangements. Microfilaments throughout the egg are identified as actin by their reversible heavy meromyosin binding. Before the second PBF, a distinct network of actin filaments is present in the endoplasmic region. It is disorganized during the second PBF; short actin filaments are caused to aggregate with membranous organelles. Following the second PBF, similar short filaments become localized in the subcortical layer but not in the underlying yolky region. However, it is not until 50-60 min after the second PBF that an elaborate actin network is established in the subcortical layer. The cortex contains a sheet-like lattice of actin filaments. It is thickest around the animal pole, and tapes toward the equator of the egg. At about 90 min after the second PBF, this polarized distribution of cortical filaments becomes more pronounced as the result of their movements. Chronologically, subcortical actin network formation and cortical reorganization correspond to the later portion of the first step and the earlier portion of the second step of ooplasmic segregation, respectively. These findings are discussed in terms of ooplasmic movements and rearrangements.     

The fate of the onychophoran antenna

Recent gene expression data suggest that the region on which the onychophoran antenna is situated corresponds to the anteriormost, apparently appendage-less region of the arthropod head. The fate of the onychophoran antenna (or any appendage-like precursor), also called the primary antenna, has been discussed intensively, and there are conflicting suggestions that this anteriormost non-segmental appendage gave rise either to the arthropod labrum or, alternatively, to the so-called frontal filaments found in certain crustaceans. Our data on early axogenesis in anostracan crustaceans show that even in the earliest embryos, before the antennula and antennal nerves are developed, the circumoral anlagen of the brain display very prominent nerves which run into the frontal filament organ (also known as the cavity receptor organ). This situation resembles the development of the antennal nerves in onychophorans, which leads us to conclude that the frontal filaments are indeed homologous to the primary antenna. Frontal filaments also appear to be more common in crustaceans than previously thought, removing the need for a complicated scenario of transformation from a primary antenna into the labrum.     

Phases of cytoplasmic and cortical reorganizations of the ascidian zygote between fertilization and first division.

Many eggs undergo reorganizations that localize determinants specifying the developmental axes and the differentiation of various cell types. In ascidians, fertilization triggers spectacular reorganizations that result in the formation and localization of distinct cytoplasmic domains that are inherited by early blastomeres that develop autonomously. By applying various imaging techniques to the transparent eggs of Phallusia mammillata, we now define 9 events and phases in the reorganization of the surface, cortex and the cytoplasm between fertilization and first cleavage. We show that two of the domains that preexist in the egg (the ER-rich cortical domain and the mitochondria-rich subcortical myoplasm) are localized successively by a microfilament-driven cortical contraction, a microtubule-driven migration and rotation of the sperm aster with respect to the cortex, and finally, a novel microfilament-dependant relaxation of the vegetal cortex. The phases of reorganization we have observed can best be explained in terms of cell cycle-regulated phases of coupling, uncoupling and recoupling of the motions of cortical and subcortical layers (ER-rich cortical domain and mitochondria-rich domain) with respect to the surface of the zygote. At the end of the meiotic cell cycle we can distinguish up to 5 cortical and cytoplasmic domains (including two novel ones; the vegetal body and a yolk-rich domain) layered against the vegetal cortex. We have also analyzed how the myoplasm is partitioned into distinct blastomeres at the 32-cell stage and the effects on development of the ablation of precisely located small fragments. On the basis of our observations and of the ablation/ transplantation experiments done in the zygotes of Phallusia and several other ascidians, we suggest that the determinants for unequal cleavage, gastrulation and for the differentiation of muscle and endoderm cells may reside in 4 distinct cortical and cytoplasmic domains localized in the egg between fertilization and cleavage.     

白额鹱卵壳的扫描电镜观察

本文报道白额鹱卵壳的壳膜、孔锥层、海绵层、表层等的超微结构,并对卵壳元素进行ＴＮ－５５００能谱分析。     

Resilience of anostracan cysts to fire

California's Mediterranean ecosystems include shrubland and grassland vegetation types that are fire-prone. Dotted within this landscape are ephemeral wetlands called vernal pools. Since surrounding upland vegetation is adapted to survive fire, it is expected that vernal pool organisms should be able to survive as well. One group of animals common to vernal pools are anostracan crustaceans that survive the pool's dry period as encysted embryos. We hydrated anostracan cysts from the soil of a recently burned pool and from soil samples intentionally burned in a prescribed fire. We also sampled burned pools when refilled the next rainy season. We found that anostracan cysts in the soil can survive fire and that shrimp occur in pools in the first post-burn season. This information is important from a management perspective concerning fire effects, controlled or natural, on vernal pools and their rare and endangered species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Origin of the electroencephalogram

An attempt is made to estimate the contribution of the activity of each cortical layer of rabbit's brain and subcortical structure in EEG formation. It has been shown that the main generator layers of spontaneous EEG are the I layer--the layer of apical dendrites and the V--the layer of pyramidal neuron somas. To perform a quantitative estimation of passive penetration of the electric fields from deep-seated structures, chiefly from the hippocampus, biopotentials studied were recorded from I, V cortex layers and hippocampus simultaneously, the cortex being disengaged by the method of propagating depression. Significant suppression of surface-activity of the motor cortex was achieved with its preservation in the hippocampus.     

Alveolar Echinococcus species from Vulpes corsac in Hulunbeier, Inner Mongolia, China, and differential development of the metacestodes in experimental rodents

Adults of alveolar Echinococcus species with different uterine structures were collected from Vulpes corsac in the Hulunbeier Pasture of Northeastern China in 2001. They were Echinococcus multilocularis Leuckart, 1863 (type No. 3, similar to E. m. multilocularis), with vaselike uterus; Echinococcus cf. sibiricensis Rausch et Schiller, 1954 (type No. 1), with pyriform uterus; and Echinococcus sp. (type No. 2) with spherical uterus at segment top. The metacestode development in rodents also differed among those 3 parasites. In the case of E. multilocularis (type No. 3), many germinal cells grew on the inner surface of early cysts, most of which metastasized into host tissue to form brood vesicles or from the germinal cell layer on the inner surface of the vesicle wall. Cells also had an appearance of proliferating by means of alveolar buds from alveolar tissue that developed outward to form new alveolar foci. In Echinococcus cf. sibiricensis (type No. 1), the formation of alveolar vesicles was due to the metastasizing of germinal tissue into host tissue; protoscoleces grew in the center of alveolar vesicles. In type No. 2 (Echinococcus sp.), the formation of the alveolar vesicle was by multiplication of germinal cell layers on the inner surface of alveolar cysts; protoscoleces grew from the germinal cell layer and mesh in the vesicles. On the basis of uterine structure and on differences in development of metacestodes in experimental rodents, we propose that the 3 types of Echinococcus represent 3 independent species: E. multilocularis, Echinococcus sibiricensis, and Echinococcus sp. (type No. 2-as yet under study).     

Airway level at which edema liquid enters the air space of isolated dog lungs

To identify lung units associated with liquid leakage into the air space in high-pressure pulmonary edema, we perfused air-inflated dog lung lobes with albumin solution to fill the loose peribronchovascular interstitium. Next, we perfused the lobes for 90 s with fluorescent albumin solution then froze the lobes in liquid nitrogen. This procedure confined the fluorescent perfusate to the liquid flux pathway between the circulation and the air space and eliminated the previously filled peribronchovascular cuffs as a source of the fluorescence that entered the air space. We divided each frozen lobe into three horizontal layers and prepared fluorescence-microscopic sections of each layer. In the most apical layers where alveolar flooding was minimal, 10.6 +/- 21.0% (SD) of alveolar ducts were either fluorescence filled or air filled and continuous with fluorescence-filled alveoli. In the same layers, 11.0 +/- 19.0% of respiratory bronchioles were similarly labeled. No terminal bronchioles in these layers were fluorescence labeled. This suggested that the fluorescent albumin entered the air space across the epithelium of respiratory bronchioles, alveolar ducts, or their associated alveoli. To simulate an alternative explanation, i.e., that fluorescence first entered central airways then flowed into peripheral air spaces, we prepared two additional lobes that we first partially inflated with fluorescent albumin then filled to capacity with air. This pushed the fluorescent solution along the airways into the lung periphery. In these lobes the ciliary lining of bronchi and terminal bronchioles was fluorescence coated. By comparison, cilia in fluorescence-perfused lobes were not coated. We conclude that alveolar flooding in hydrostatic pulmonary edema occurs across the epithelium of alveolar ducts, respiratory bronchioles, or their associated alveoli.(ABSTRACT TRUNCATED AT 250 WORDS)     

Light microscopy of the medial wall of the cerebral cortex of the lizard Psammodromus algirus

The telencephalic medial wall of the lizard Psammodromus algirus was studied using Golgi and conventional light microscopic techniques. The area is formed by two different cytological fields—medial cortex and dorsomedial cortex. These two cortices possess three layers dorsoventrally: a superficial plexiform layer, a cellular layer, and a deep plexiform layer. The alveus, a deep fiber system, runs adjacent to the ependyma. Four classes of neurons are found in the cellular layer of the medial cortex on the basis of soma shape, dendritic pattern, and position in the layer: horizontal, double pyramidal, and candelabra cells. Solitary cells are present in the superficial and deep plexiform layers of the medial cortex. Those of the superficial plexiform layer are stellate cells. Horizontal and vertical cells are found in the deep plexiform layer. Double pyramidal cells are the most frequently impregnated in the cellular layer of the dorsomedial cortex. In addition, candelabra cells are present at the lateral end of the layer. Two cell types are found in the deep plexiform layer of the dorsomedial cortex: solitary pyramidal cells and, among the fibers of the alveus, horizontal cells. Ependymal tanycytes line the ventricular surface, and protoplasmic astrocytes are found in the plexiform layers of both medial and dorsomedial cortices.