Embryogenesis in mouth-breeding cichlids (Osteichthyes,Teleostei) structure and fate of the enveloping layer

The eggs of African mouth-brooders are of unusual size and shape. Studying their development may help to more clearly understand epiboly, gastrulation, and the relation between enveloping layer (periderm) and epidermis. When epiboly has progressed over just one fifth of the yolk mass, the germ ring and embryonic shield are already well established. Behind the germ ring very few deep cells are present at this early stage of epiboly, except in the embryonic shield. When the blastodisc covers the animal half of the yolk mass, the future body is already well established with notochord, somites and developing neural keel. Apart from these structures, no deep cells can be detected between enveloping layer and yolk surface; not even a germ ring remains behind the advancing edge of the enveloping layer. Epiboly over the greater part of the yolk is achieved only by the enveloping layer and the yolk syncytial layer. As the margin of the enveloping layer begins to reduce its circumference when closing around the vegetal pole, groups of cells in the advancing edge become spindle-shaped, with a single cell in between of each of these groups broadening along the edge. The enveloping layer (called periderm after epiboly) remains intact until after hatching, when, together with the underlying ectoderm, it forms the double-layered skin of the larval fish. Thereafter, cells deriving from the subperipheral ectoderm gradually replace the decaying periderm cells to form the final epidermis. Thus, in the cichlids studied, the enveloping layer alone forms the yolk sac to begin with, and it covers the larval body until some days after hatching.     

Differentiation of swimming muscles and gills,and development of anaerobic power in the larvae of cyprinid fish (Pisces,Teleostei)

Summary Histological, histochemical, morphometric and electrophoretic methods were combined to study the differentiation of the swimming muscles and of the gills in the larvae and juveniles of cyprinids during the first 3 months after hatching. The bodies of recently hatched larvae are always surrounded by a single layer of muscle fibres (red layer) which possess strong cytochrome oxidase (COX) activity but whose myofibrils do not show the arrangement typical of the fibres of adult red muscle. The contribution of the red layer to total muscle mass decreases from about 12% on day 3 to 4% on day 40 post-hatching, after which the red layer becomes indistinguishable from the developing mass of adult red muscles. The adult fibres seem to originate through splitting from the larval fibres. The inner muscle masses of recently hatched larvae also display conspicuous COX activity which, however, disappears gradually. The development of the gill surface follows a time course which during the first 40 days is a mirror image of that of the larval red layer of muscle fibres, i.e. increasing at about the same rate as the latter decreases. The most cathodic of the isoenzymes of LDH (M₄), an indicator of the glycolytic capacity of the white muscle of adult fish, develops only slowly in the larvae. In whole body homogenates, the dominant isoenzyme after hatching is the aerobic H₄-form, and it is not until 2 weeks or so that the anaerobic M₄ is the strongest fraction in electropherograms of total body homogenates. In the bleak, Alburnus alburnus (Linné, 1758), which spends the first 8–10 days after hatching on the bottom of the aquarium, the M₄ isoenzyme takes much longer to develop than in Rutilus rutilus (Linné, 1758) or Chondrostoma nasus (Linné, 1758) which start swimming within 3 days after hatching. All these findings suggest that in the early larvae swimming is almost entirely aerobic, being powered by the deep layers of muscle fibres. During this developmental phase the superficial red layer perhaps is the main respiratory organ.     

PACAP in developing sensory and peripheral organs of the zebrafish, Danio rerio

The anatomical distribution of PACAP-like immunoreactivity was investigated in sensory and peripheral organs of the zebrafish, Danio rerio, during the pharyngula, hatching and larval periods, by using indirect immunofluorescence methods. First PACAP-like immunoreactive (ir) elements appeared during the pharyngula period, at 24 hours post fertilization (hpf), within the most superficial layer of the retina and the dorsal aorta. At 48 hpf, additional ir cells were found in the olfactory placode and esophagus. At 72 hpf (hatching period), PACAP-like immunoreactivity was first detected in the ganglion cell layer of the retina, the otic sensory epithelium, pharyngeal arches, swim bladder and pancreatic progenitor cells. During day 5 of larval development, new groups of ir cells appeared in the liver, whereas no ir elements were observed in the olfactory placode. Subsequently, at day 13 of larval development, additional ir elements were found for the first time in some gut epithelial cells while those previously observed in the retina and otic sensory epithelium were absent. The transient expression of PACAP-like ir material in sensory organs suggests that the peptide could be implicated in neurotrophic activities and neurosensorial connections in the migration and/or differentiation processes. The appearance of PACAP-like ir elements in peripheral organs at different developmental stages, indicates that this peptide could be involved in the control of more specific functions as soon as these peripheral structures begin to operate.     

Minute tubercles on the skin surface of larvae in the Korean endemic bitterling, Rhodeus pseudosericeus (Pisces, Cyprinidae)

The morphology and distribution of the minute tubercles on the skin surface of larvae in Korean bitterling, Rhodeus pseudosericeus, were observed during larval development. Just after hatching, the epidermis of the larvae consists of a thin single cell layer having smaller basophilic flat or round‐flattened basal cells. As the larvae grow, the epidermis contains more small flat cells and large epidermal cells that are round or hemisphere‐shaped. These large unicellular epidermal cells, called minute tubercles, consist of more or less homogeneous cytoplasm that is PAS (Periodic acid‐Schiff method) positive. They are more densely distributed in the wing‐like yolk sac projection. Vestigial minute tubercles occur in the body region and the caudal fin‐fold region. These minute tubercles grow in number and height from 6 to 8 days after hatching onward. However, they become reduced in height and number as the larvae develop. At day 31 after hatching (i.e. free‐swimming stage), minute tubercles no longer exist on the larval skin. The sequence of occurrence and gradual disappearance of these cell structures are described and histologically documented for comparative purposes of beta, taxnomomic and environmental studies.     

Salivary gland development in the blowfly,Calliphora erythrocephala

The salivary gland of adult Calliphora erythrocephala is a tubular structure composed of secretory, reabsorptive, and duct regions. Development of these structures has been followed during the six days of larval and ten days of pupal growth. Two small groups of imaginal cells located at the junction between larval gland and duct give rise to the adult gland. These presumptive adult cells divide during all larval stages and appear to be functional components of the larval gland. Shortly after pupation, the larval gland breaks down and the imaginal cells proliferate rapidly, forming sequentially the duct, reabsorptive and secretory regions. Proliferating regions of the developing gland are frequently encrusted with haemocytes. As it elongates the gland establishes intimate contacts first with the basement membrane of the degenerating larval gland, later with an epithelial layer surrounding the main dorsal tracheal trunks, and then with the gut. Cell division continues until about five days after pupation, bu t the gland is unable to secrete fluid in response to 5-hydroxytryptamine stimulation until two hours after the adult fly emerges. The Golgi complex appears to be involved in forming the highly folded membranes of the canaliculi in the secretory region. Presumptive adult salivary gland cells appear to increase in number logarithmically from the time of hatching of the larva until five days after pupation. This contrasts with the development of classical imaginal discs, in which cell division ceases prior to pupation.     

Life styles and patterns of development of gills and muscles in larval cyprinids (Cyprinidae; Teleostei)

A morphometric study of gill structures and of the body musculature during the first weeks after hatching was carried out on larvae of six cyprinid species: Leuciscus cephalus, L. leuciscus, Rutilus rutilus, Alburnus albumus, Chondrostoma nasus and Abramis brama . In all species a unicellular layer of red muscle fibres covers the central muscle mass; this layer is of greatest extent shortly after hatching but diminishes gradually in mass by contracting towards the lateral region of the body until it merges with (or gives rise to) the adult red muscle fibres proper. There is a close relationship between the rate of differentiation of gill structures and the rate at which the larval red muscle layer disappears, the pattern of this relationship reflecting the life style of the species. The longer the larvae delay the start of their free-swimming existence after hatching (which in A. alburnus may be as long as 10 days) the longer does the red layer of muscle fibres serve as the organ of gas exchange and the longer is gill development suppressed. It appears that the metabolism of the swimming muscles is almost entirely aerobic so long as gas exchange takes place across the whole body surface, the glycolytic capacity of the central muscle mass developing only slowly in conjunction with the switch from red layer to gills as the major respiratory organ.     

The role of intragemmular osmotic pressure in cell division and hatching of gemmules of the fresh-water sponge Spongilla lacustris (Porifera)

Summary The gemmule coat of Spongilla lacustris is histologically single-layered in the gemmules studied in this work. This single layer is comparable to the classically described internal chitinous membrane of Leveaux (1939). It has been found to contain collagen with an axial period in electron micrographs of about 120 Å and is bounded internally by a thin dense layer which is separate from the internal gemmular cells, and which may be chitinous.Gemmules of this sponge studied during March to June of 1973 respond to 230 mOsmolar solutions of small molecules by: 1. undergoing no change, in which case the substances are freely permeable to the gemmule coat and cells; 2. displaying shrinkage of the cell mass, in which case the substances are permeable to the coat but relatively impermeable to the cells; 3. displaying folding of the coat and cell mass shrinkage because the substances are relatively impermeable to both the coat and the cells; and 4. displaying complete collapse of the gemmule due to impermeability to the coat. The lipid solubility of a substance is directly related to its ability to penetrate the coat. Further, molecular size and charge are also of apparent importance.Substances which penetrate the coat and remain osmotically active (are not metabolized) inhibit hatching. Low concentrations of sodium chloride (23 mOsmolar) have been demonstrated to reversibly inhibit hatching. Higher concentrations cause irreversible damage at 20° C but have little effect at 4° C, indicating that damage is related to the metabolic level of the cells. Once hatching is stimulated by increased temperature the cells become progressively less sensitive to an increase in osmotically active substances.Inhibition of gemmule hatching can theoretically occur by: 1. an addition of solutes to the gemmular fluid, or 2. through an increase in concentration of intragemmular solutes by water withdrawal.Our results raise the question of whether the inhibition of hatching by gemmulostasine, reported by Rasmont (1965) and Rozenfeld (1970, 1971), is due to an osmotic effect rather than to a specific physiological one.Based upon the results reported here and on the work of Zeuthen (1939) and Schmidt (1970) we propose a tight coupling between the intragemmular osmotic pressure and the triggering of hatching (cell division). Any substance which increases intragemmular osmotic pressure to a large enough extent will inhibit hatching. Furthermore, it can be hypothesized that hatching is normally triggered by a decrease in osmotic pressure due to water movement into the gemmule, the movement of solutes out of the gemmule, or to a combination of these.This work was supported by a grant from the National Science Foundation (GB-37775) to T. L. S.     

Demonstration of the granular layer and the fate of the hyaline layer during the development of a sea urchin (Lytechinus variegatus)

Summary Employing electron-microscopic methods that help retain polyanionic materials, we describe the extracellular coverings of a sea urchin (Lytechinus variegatus) throughout ontogeny. The surface of the embryo is covered by a two-layered cuticle (commonly called the hyaline layer), which in turn is covered by a granular layer. The granular layer is retained after addition of alcian blue to the fixative solutions, and has not been previously described for any sea urchin. After hatching, the granular layer disappears, but the hyaline layer continues to cover most of the larval surface until settlement and metamorphosis. A few days before metamorphosis, the hyaline layer lining the vestibular invagination of the competent pluteus larva is replaced by a three-layered cuticle resembling that of the adult sea urchin. The hyaline layer covering the rest of the larva is evidently lost at metamorphosis during the involution of the general epidermis.     

Growth and respiration of embryos and larvae of the rabbittish Siganus randalli (Pisces, Siganidae)

Growth and respiration of larval rabbitfish from Guam were examined. Larvae were reared from eggs in 2- to 10-ton tanks and were fed rotifers, Anemia , and artificial feed in succession as development proceeded through metamorphosis. Growth in length was rapid during the 12 h after hatching, then slowed until the larvae began to feed. The yolk sac was usually absorbed by 36 h after hatching. Rates of respiration of larvae and eggs were determined with a dissolved oxygen electrode at various times through development. Larval metabolism increased steadily during the embryonic stages culminating in a metabolic burst immediately after hatching. Respiration rates remained relatively stable from shortly after hatching until the onset of exogenous feeding, after which respiration rates increased with larval size. The respiration rates of post-yolk-sac larvae scaled isometrically with larval dry mass. Daily growth of feeding larvae was 27 to 28% of larval dry mass.     

Functional development in the mauthner cell system of embryos and larvae of the zebra fish

In the embryonic zebra fish as early as 40 hr after fertilization, the Mauthner cells (M-cells) initiate an escape response, elicited by tactile-vibrational stimulation. The initial part of this behavior is similar to the acoustic startle reflex seen during the larval stage which begins at 96 hr. The embryonic response is directional and is followed by a series of strong tail flexures which are more pronounced than those during swimming. In the embryo the M-cell fired at the beginning of the response and rarely fired again during subsequent contractions; in our experiments the M-cell did not mediate iterative movements of the tail. The M-cell system is probably involved in evoked hatching behavior, as the tactile response is sufficient to rupture the egg membrane and allow the animal to escape. The M-cell sometimes fired spontaneously, which suggests that it might function also in spontaneous hatching behavior which occurs in the absence of phasic stimulation. At 48 hr the M-cell has morphologically mature synapses on its soma and dendrites, but its cytoplasm is relatively undifferentiated; it has few oriented neurofilaments and no distinct axon hillock. During these stages the extracellular M-spike is longer in duration and smaller in amplitude than at later times when the cell is more mature morphologically. Our data suggest that long-term inhibitory control of the M-cell system begins to function at about the time of hatching. At this time the cell is morphologically mature and is richly supplied with synaptic endings over its soma and dendrites.     

自然干燥对冬虫夏草寄主蝠蛾卵孵化的影响

研究在室内自然空气湿度下放置的时间长短对冬虫夏草(Cordyceps)寄主昆虫贡嘎蝠蛾Hepialus gonggaensis Fu et Huang卵孵化率的影响。卵早期的研究结果为:第1批、第2批和第3批卵于室内自然空气湿度下保存的时间达26,11和16h后再保湿都可以正常孵化并且孵化率与对照无显著性差异,所孵化幼虫在饲养初期的成活率分别达62.0%,41.4%和43.4%,与对照无显著性差异。卵中期干燥放置36h的孵化率为66.7%,所孵化幼虫在饲养初期的成活率为50.0%,孵化率和成活率都与对照无显著性差异。卵晚期干燥放置24h的孵化率为70.0%,所孵化幼虫在饲养初期的成活率为49.0%,孵化率和成活率都与对照无显著性差异。以上结果表明,经历一定时间的干燥不会对卵的正常孵化有影响。     

Variation in the hatching behaviour of Nematodirus battus: Polymorphic bet hedging?

Previous work on the transmission dynamics of Nematodirus battus, an important nematode parasite of farmed ruminants in temperate regions, suggests that it operates a bet-hedging strategy. Hatching of cold-sensitised eggs is concentrated in spring, while alternative hatching of non-cold-sensitised eggs in autumn mitigates the risk of poor conditions for hatching in spring or host absence during peak larval availability. Isolates from Scotland showed much less propensity to hatch without chilling than the previously characterised isolate from southern England. Nematodirus battus eggs from a hill farm in Scotland showed intermediate proportions of non-chilled hatching, perhaps related to unpredictability of climate at higher altitudes. Geographic polymorphism in larval behaviour appears to be present in the form of differing chilling requirements for egg hatching. Since bet-hedging through trait diversification is a plausible and demonstrated strategy for coping with environmental unpredictability, it is a likely target for adaptation to climate change. Predictions of disease epidemiology in a changing climate should incorporate parasite adaptation, but further theoretical and empirical characterisations of likely evolutionary responses are needed before this is possible for the most economically important systems.     

Induction of hatching by chemical signals secreted by the ovigerous female of an estuarine crab Sesarma haematocheir

Hatching of embryos in the estuarine crab Sesarma haematocheir is highly synchronized with nocturnal high tide and completes within 1 hr among all embryos incubated by the female. This highly synchronized hatching is induced by a "Hatching-Program Inducing Factor (HPIF)" released from the female. To further define the cues involved in synchronized hatching, experiments were designed to characterize this factor and to determine possible sites of release and temporal release patterns using strategies involving isolation of egg masses, perfusion, and ablation experiments on fully developed embryos that had not yet entered the hatching program. Embryo transplantations indicate that not only HPIF may be released from the branchial chamber, but that it is extraordinarily unstable, and loses activity within 15 min, which frustrates further attempts at characterization. Nevertheless, with regard to temporal release patterns, it was established that HPIF activity was detected during short periods over three consecutive nights prior to release of larvae. This activity did not explain the gated response of embryo release in this crab, which might correspond with circatidal larval release events in the field.     

Evolutionary modification of mesenchyme cells in sand dollars in the transition from indirect to direct development

Peronella japonica, an intermediate type of direct-developing sand dollar, forms an abbreviated pluteus, followed by metamorphosis within 3 days without feeding. In this species, ingression of mesenchyme cells starts before hatching and continues until gastrulation, but no typical secondary mesenchyme cells (SMCs) migrate from the tip of the archenteron. Here, I investigated the cell lineage of mesenchyme cells through metamorphosis in P. japonica and found that mesenchyme cells migrating before hatching (early mesenchyme cells [EMCs]) were exclusively derived from micromeres and became larval skeletogenic cells, whereas cells migrating after hatching (late mesenchyme cells [LMCs]) appeared to contain several nonskeletogenic cells. Thus, it is likely that EMCs are homologous to primary mesenchyme cells (PMCs) and LMCs are similar to the SMCs of typical indirect developers, suggesting that heterochrony in the timing of mesenchyme cell ingression may have occurred in this species. EMCs disappeared after metamorphosis and LMCs were involved in adult skeletogenesis. Embryos from which micromeres were removed at the 16-cell stage formed armless plutei that went on to form adult skeletons and resulted in juveniles with normal morphology. These results suggest that in P. japonica, LMCs form adult skeletal elements, whereas EMCs are specialized for larval spicule formation. The occurrence of evolutionary modifications in mesenchyme cells in the transition from indirect to direct development of sand dollars is discussed.     

柑桔爆皮虫幼期各虫态的形态学观察

采用田间观察和室内观察相结合的方法,使用Leica MZ 16A研究型体视解剖镜,对柑桔爆皮虫Agrilus auriventris Saunders幼期各虫态的形态进行了系统的观察。结果发现：卵初产时为乳白色,后变为浅褐色并开始皱缩,末期变为黄褐色,孵化前部分卵壳分离为白色蜡质外层和黄褐色内层两层;幼虫孵化的同时直接从卵壳贴近树皮的一侧蛀入韧皮部,同时将大量虫粪排入卵壳;幼虫随着虫龄的增加钳状突的阶数和口上片的宽度发生有规律的变化,末龄幼虫在木质部作一椭圆形蛹室化蛹,身体的头部和尾部以3∶7的比例对折; 化蛹前的幼虫经历一个身体缩短的预蛹状态;刚化的蛹为乳白色,复眼颜色与身体相似,化蛹后蛹的颜色逐渐变为淡黄色,复眼颜色变为浅红褐色,后期复眼变为红褐色,口器变黑,羽化前蛹的复眼、口器和体表全部变黑;羽化过程中,成虫体表的膨胀导致蛹表皮脱裂,前翅变黑和腹部背面变成碧蓝色后羽化成完整的成虫。本结果进一步支持将柑桔爆皮虫幼虫期分为5龄。     

Structure and development of spines over the skin surface of the river puffer Takifugu obscurus (Tetraodontidae,Teleostei) during larval growth

The histological structure and development of spines on the skin surface of Takifugu obscurus were studied during larval development conducted artificially with an average 30‰ salinity and 18.0–20.3°C water temperature. The epidermis comprises an outermost layer, middle layer, and the stratum germinativum, and contains three types of gland cells: small spherical or flask‐shaped mucous cells, larger sacciform mucous cells, and large granular cells. The dermis and subcutis follow. The spines first appear over the ventral region at 10 days after hatching and consist of two parts: a central long tapering portion which projects into the epidermis and eventually outside of the body, and a short supporting basal portion that is embedded within the stratum compactum layer of the dermis. The central, long tapering portion has two very short processes on top until 25 days after hatching, but these two separate spines fuse into one 30 days after hatching. In contrast, the short supporting spines rooted at the base consist of three to six small spines (usually four to five spines) and are present even in the adult stage. Therefore, calcareous spines consisting of one central long spine and three to six smaller supporting spines form tetra‐ and septaradiate spines (mainly penta‐ and hexaradiate). The spines first appear over the ventral region.     

Ultrastructure of larval epidermis of the nemertean <Emphasis Type="Italic">Quasitetrastemma stimpsoni</Emphasis> (Chernyshev, 1992) (Hoplonemertea)

The ultrastructure of the epidermis of free-swimming larvae of the nemertean Quasitetrastemma stimpsoni was examined. At about 24 hours after hatching, the provisional epithelium of larva is 28–35 μm thick and consists of two layers of cells—peripheral and basal. The peripheral layer consists of multiciliated cells and two kinds of gland cells. The “basal cup” zone is formed from the vacuoles of basal cells. At about 50 hours after hatching, the definitive epithelium is 14–17 μm thick and exhibits a typical hoplonemertean structure. However, it has numerous yolk vesicles, few processes of granular basal cells, and a weakly developed dermis. Thus, the replacement of the provisional epithelium by the definitive one occurs in Q. stimpsoni at an earlier stage, compared to the hidden larva of Tetrastemma candidum.     

The central nervous system of the ascidian larva: mitotic history of cells forming the neural tube in late embryonic Ciona intestinalis

Ascidian larvae develop after an invariant pattern of embryonic cleavage. Fewer than 400 cells constitute the larval central nervous system (CNS), which forms without either extensive migration or cell death. We catalogue the mitotic history of these cells in Ciona intestinalis, using confocal microscopy of whole-mount embryos at stages from neurulation until hatching. The positions of cells contributing to the CNS were reconstructed from confocal image stacks of embryonic nuclei, and maps of successive stages were used to chart the mitotic descent, thereby creating a cell lineage for each cell. The entire CNS is formed from 10th- to 14th-generation cells. Although minor differences exist in cell position, lineage is invariant in cells derived from A-line blastomeres, which form the caudal nerve cord and visceral ganglion. We document the lineage of five pairs of presumed motor neurons within the visceral ganglion: one pair arises from A/A 10.57, and four from progeny of A/A 9.30. The remaining cells of the visceral ganglion are in their 13th and 14th generations at hatching, with most mitotic activity ceasing around 85% of embryonic development. Of the approximately 330 larval cells previously reported in the CNS of Ciona, we document the lineage of 226 that derive predominantly from A-line blastomeres.     

Growing pains: development of the larval nocifensive response in Drosophila

The ability to perceive and avoid harmful substances or stimuli is key to an organism's survival. The neuronal cognate of the perception of pain is known as nociception, and the reflexive motion to avoid pain is termed the nocifensive response. As the nocifensive response is an ancient and evolutionarily conserved behavioral response to nociceptive stimuli, it is amenable to study in relatively simple and genetically tractable model systems such as Drosophila. Recent studies have taken advantage of the useful properties of Drosophila larvae to begin elucidating the neuronal connectivity and molecular machinery underlying the nocifensive response. However, these studies have primarily utilized the third-instar larval stage, and many mutations that potentially influence nociception survive only until earlier larval stages. Here we characterize the nocifensive responses of Drosophila throughout larval development and find dramatic changes in the nature of the behavior. Notably, we find that prior to the third instar, larvae are unable to perform the characteristic "corkscrew-like roll" behavior. Also, we identify an avoidance behavior consistent with a nocifensive response that is present immediately after larval hatching, representing a paradigm that may be useful in examining mutations with an early lethal phenotype.     

The ecological and anatomy of a new species of aeolid opisthobranch mollusc; a predator of the scleractinian coral Porites

A new species of aeolid opisthobranch (family Tergipedidae) is described from Tanzania. It lives and feeds on the scleractinian coral Porites somaliensis. Studies on its development show that it can reach sexual maturity three weeks after hatching. The larval stage is extremely abbreviated; the veliger settles ten minutes after hatching, and within ninety minutes of hatching has assumed a slug-like form.