版纳鱼螈幼体和成体皮肤的结构

采用光镜和扫描电镜,对我国特有的珍稀濒危两栖动物版纳鱼螈Ichthyophis bannanicus幼体和成体的皮肤进行形态学和组织学观察.版纳鱼螈幼体和成体的皮肤均可分为表皮、真皮疏松层和真皮致密层;皮肤中含有粘液腺和颗粒腺;在不同发育阶段或同一个体的不同部位,其皮肤的各种组成成分在结构和厚度上存在着差异:成体和幼体都是头部的表皮最厚,尾部的最薄;幼体表皮各层细胞分化不明显,几无角化现象,成体表皮的各层细胞分化明显,表层细胞明显角化;成体躯干部皮肤最厚,头部最薄,幼体则是头部皮肤最厚,尾部最薄;幼体和成体的头部皮肤都分布有大量的粘液腺,颗粒腺分布较少;幼体的躯干部皮肤则主要分布着大量颗粒腺,尾部只有颗粒腺,未见粘液腺;成体躯干部和尾部皮肤均分布有大量的颗粒腺和粘液腺.     

Two Transitions of Haemoglobin Expression in Xenopus: from Embryonic to Larval and from Larval to Adult

Abstract. Electrophoretic analyses of haemoglobin and globin phenotypes in families of Xenopus borealis and Xenopus l. laevis revealed two developmental haemoglobin transitions during ontogeny. The first transition occurs at the developmental stage when tadpoles begin to feed. It is characterized by the decline of embryonic-specific globins in favour of novel, tadpole-specific globins ( X. borealis ) correlated to changes in the haemoglobin pattern. We suppose that this switch results from the replacement of a primitive, ventral blood island-dependent erythrocyte population by tadpole erythrocytes from other erythropoietic sites. Several other globin chains and haemoglobins are present in both young tadpoles and throughout larval life. The second, well-known transition occurs during metamorphosis, where all tadpole haemoglobins are replaced by adult haemoglobins composed of entirely different globin chains.     

Heart Dissection in Larval,Juvenile and Adult Zebrafish,Danio rerio

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews^1-6), however, work examining post-embryonic through adult cardiac development has been limited^7-10. Examining the changing morphology of the maturing and aging heart are restricted by the lack of techniques available for staging and isolating juvenile and adult hearts. In order to analyze heart development over the fish''s lifespan, we dissect zebrafish hearts at numerous stages and photograph them for further analysis¹¹. The morphological features of the heart can easily be quantified and individual hearts can be further analyzed by a host of standard methods. Zebrafish grow at variable rates and maturation correlates better with fish size than age, thus, post-fixation, we photograph and measure fish length as a gauge of fish maturation. This protocol explains two distinct, size dependent dissection techniques for zebrafish, ranging from larvae 3.5mm standard length (SL) with hearts of 100μm ventricle length (VL), to adults, with SL of 30mm and VL 1mm or larger. Larval and adult fish have quite distinct body and organ morphology. Larvae are not only significantly smaller, they have less pigment and each organ is visually very difficult to identify. For this reason, we use distinct dissection techniques.We used pre-dissection fixation procedures, as we discovered that hearts dissected directly after euthanization have a more variable morphology, with very loose and balloon like atria compared with hearts removed following fixation. The fish fixed prior to dissection, retain in vivo morphology and chamber position (data not shown). In addition, for demonstration purposes, we take advantage of the heart (myocardial) specific GFP transgenic Tg(myl7:GFP)^{twu34 (12)}, which allows us to visualize the entire heart and is particularly useful at early stages in development when the cardiac morphology is less distinct from surrounding tissues. Dissection of the heart makes further analysis of the cell and molecular biology underlying heart development and maturation using in situ hybridization, immunohistochemistry, RNA extraction or other analytical methods easier in post-embryonic zebrafish. This protocol will provide a valuable technique for the study of cardiac development maturation and aging.Download video file.^{(42M, mov)}     

The nervus rarus in Coelacanthiform Phytogeny

Bjerring, H. C. (Section of Palaeozoology, Swedish Museum of Natural History, Stockholm, Sweden). The nervus rarus in coelacanthiform phylogeny. Zool. Scripta 1(2): 57–68, 1972.–In the course of craniate vertebrate evolution the somatic motor portion of the second cranionerval segment probably became divided into two nerves, viz. the trochlear nerve and the rarus nerve. The rarus nerve, allied to the second-metamere basicranial muscle, apparently migrated backwards from the middle to the posterior part of the evolving head. Devonian coelacanthiforms exhibit an endocranial passage indicating that the rarus nerve accompanied the glossopharyngeal nerve through the otic capsule; in contrast, in the Recent Latimeria, the corresponding nerve emerges from the endocranium somewhat further posteriorly in association with the vagus nerve. Within the evolutionary history of the coelacanthiforms, this backward displacement of the rarus nerve appears to be correlated with a progressive development of the second-metamere basicranial muscle as well as with a number of changes in both the head skeleton and the brain.     

Dissection and Immunohistochemistry of Larval,Pupal and Adult Drosophila Retinas

The compound eye of Drosophila melanogaster consists of about 750 ommatidia (unit eyes). Each ommatidium is composed of about 20 cells, including lens-secreting cone cells, pigment cells, a bristle cell and eight photoreceptors (PRs) R1-R8 ². The PRs have specialized microvillar structures, the rhabdomeres, which contain light-sensitive pigments, the Rhodopsins (Rhs). The rhabdomeres of six PRs (R1-R6) form a trapezoid and contain Rh1 ^{3 4}. The rhabdomeres of R7 and R8 are positioned in tandem in the center of the trapezoid and share the same path of light. R7 and R8 PRs stochastically express different combinations of Rhs in two main subtypes⁵: In the ''p'' subtype, Rh3 in pR7s is coupled with Rh5 in pR8s, whereas in the ''y'' subtype, Rh4 in yR7s is associated with Rh6 in yR8s ^{6 7 8}.Early specification of PRs and development of ommatidia begins in the larval eye-antennal imaginal disc, a monolayer of epithelial cells. A wave of differentiation sweeps across the disc⁹ and initiates the assembly of undifferentiated cells into ommatidia^10-11. The ''founder cell'' R8 is specified first and recruits R1-6 and then R7 ^12-14. Subsequently, during pupal development, PR differentiation leads to extensive morphological changes ¹⁵, including rhabdomere formation, synaptogenesis and eventually rh expression.In this protocol, we describe methods for retinal dissections and immunohistochemistry at three defined periods of retina development, which can be applied to address a variety of questions concerning retinal formation and developmental pathways. Here, we use these methods to visualize the stepwise PR differentiation at the single-cell level in whole mount larval, midpupal and adult retinas (Figure 1).     

Ontogeny, Phytogeny, and Cellular Cooperation

The capacity of adult newt (Triturus viridescens) spleen cellsto secrete antibody at 4 C allows simultaneous visualizationand quantification of non-secretory (S^–) and secretory(S⁺) rosette forming cells (RFC). Visualization of mammalianS⁺ RFC requires 37 C, a temperature at which S^– RFC appearto be fragile. RFC can be distinguished as S^– or S⁺ dueto whether one or more layers of erythrocytes adhere to thesurface of sensitized spleen cells. Different doses of horseerythrocytes (HRBC) affect newt S^– RFC and S⁺ RFC differentially.By varying the time between injections of different concentrationsof chicken erythrocytes (CRBC, the "carrier") and a constantdosage of CRBC-TNP (trinitrophenyl, the hapten) it is possibleto measure "helper" activity that correlates with the populationof S^– RFC and is both dose and time dependent. By varyingassay time after "helper" activity has been maximized, one candetermine the cytodynamics of anti-TNP antibody producing cell(APC) activity. For the first time these morphologically separableRFC can be related to their suspected physiologic behavior.A shift from S^– RFC to S⁺ RFC takes place during the immuneresponse. That similar dose-dependent response curves can beshown in adultRana pipiens suggests that the newt responsesrepresent a fundamental vertebrate pattern.     

Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS

The Gal4/ UAS binary method is powerful for gene and neural circuitry manipulation in Drosophila. For most neurobiological studies, however, Gal4 expression is rarely tissue-specific enough to allow for precise correlation of the circuit with behavioral readouts. To overcome this major hurdle, we recently developed the FINGR method to achieve a more restrictive Gal4 expression in the tissue of interest. The FINGR method has three components: 1) the traditional Gal4/UAS system; 2) a set of FLP/FRT-mediated Gal80 converting tools; and 3) enhancer-trap FLP (ET-FLP). Gal4 is used to define the primary neural circuitry of interest. Paring the Gal4 with a UAS-effector, such as UAS-MJD78Q or UAS-Shi^ts, regulates the neuronal activity, which is in turn manifested by alterations in the fly behavior. With an additional UAS-reporter such as UAS-GFP, the neural circuit involved in the specific behavior can be simultaneously mapped for morphological analysis. For Gal4 lines with broad expression, Gal4 expression can be restricted by using two complementary Gal80-converting tools: tub^P>Gal80> (''flip out'') and tub^P>stop>Gal80 (''flip in''). Finally, investigators can turn Gal80 on or off, respectively, with the help of tissue-specific ET-FLP. In the flip-in mode, Gal80 will repress Gal4 expression wherever Gal4 and ET-FLP intersect. In the flip-out mode, Gal80 will relieve Gal4 repression in cells in which Gal4 and FLP overlap. Both approaches enable the restriction of the number of cells in the Gal4-defined circuitry, but in an inverse pattern. The FINGR method is compatible with the vast collection of Gal4 lines in the fly community and highly versatile for traditional clonal analysis and for neural circuit mapping. In this protocol, we demonstrate the mapping of FLP expression patterns in select ET-FLPx2 lines and the effectiveness of the FINGR method in photoreceptor cells. The principle of the FINGR method should also be applicable to other genetic model organisms in which Gal4/UAS, Gal80, and FLP/FRT are used.     

Phytogeny of Epidermophyton floccosum and other dermatophytes

Eleven strains of Epidermophyton floccosum were compared with 5 Microsporum and 5 Trichophyton species with respect to the restriction fragment length polymorphism (RFLP) of the mitochondrial DNA to reveal their phylogenetic relationships. The phylogeny of 11 species showed that the three dermatophyte genera could not be separated from each other and could be considered to be congeneric. This result is not inconsistent with the results from ribosomal RNA sequences.     

Phytogeny, Variation, and Morphological Integration

Studies of intra and interspecific variability of urodele mesopodiahave revealed biases in the production of novel phenotypes;observed patterns of variability represent a small subset ofthe possible arrangements. The major trend in the phylogeneticdiversification of the salamander limb is the reduction or lossof carpal or tarsal elements, and these patterns are often highlyhomoplastic. Mesopodial arrangements that are homoplastic arecommonly encountered in studies of intrapopulational variation.Patterns of variation, when integrated with developmental biology,inform about the relationships between morphological integrationand homology. The extensive conservation of primitive developmentaland genetic interactions suggest that the individuation of structuresmaynot be complete and that homologies, themselves, may be partial.     

Differential Expression of Hox and Notch Genes in Larval and Adult Stages of Echinococcus granulosus

This investigation aimed to evaluate the differential expression of HoxB7 and notch genes in different developmental stages of Echinococcus granulosus sensu stricto. The expression of HoxB7 gene was observed at all developmental stages. Nevertheless, significant fold differences in the expression level was documented in the juvenile worm with 3 or more proglottids, the germinal layer from infected sheep, and the adult worm from an experimentally infected dog. The notch gene was expressed at all developmental stages of E. granulosus; however, the fold difference was significantly increased at the microcysts in monophasic culture medium and the germinal layer of infected sheep in comparison with other stages. The findings demonstrated that the 2 aforementioned genes evaluated in the present study were differentially expressed at different developmental stages of the parasite and may contribute to some important biological processes of E. granulosus.     

Studies on the Erythrocytes of Larval and Adult Lampreys (Lampetra fluviatilis)

The erythrocytes of larval and adult Lampetra fluviatilis were examined in suspensions of whole blood, solutions of various ionic strengths, scanning electron photomicrographs and air dried smears. Although a wide range of shapes were observed in hyper- and hypotonic solutions, the cells in whole blood and isotonic solutions were predominantly biconcave with an eccentric nucleus. The biconcave shape was also seen in scanning electron photomicrographs of erythrocytes fixed with glutaraldehyde in lamprey Ringer. The mean diameter of larval and adult cells in whole blood suspensions was 11.2 and 11.6 μm respectively. Although in air dried smears the areas of the larval and adult erythrocytes were virtually identical, the erythrocyte nuclear area was markedly higher in ammocoetes with only a small amount of overlap between the values obtained for the two stages in the life cycle.     

Adult Responses to Larval Population Size in the Almond Moth, Cadra cautella

Juvenile population size may affect the potential for future mating opportunities and therefore potentially sperm competition; this may favour ontogenetic adjustments in sperm production. Theory predicts that males should optimize their ejaculatory investment in accordance with the risk of sperm competition. Evidence for these theories is typically revealed in males of highly polyandrous species. Whether such responses to environmental cues exist for females, or are maintained in mildly polyandrous species in which most females do not re-mate, is unknown. Male lepidopterans produce normal, fertilizing sperm (eupyrene) and non-fertilizing (apyrene) sperm. Apyrene sperm are associated with reduced female receptivity, suggesting a role in sperm competition. We tested the effect of juvenile population size on life-history parameters and reproductive investment in the mildly polyandrous almond moth, Cadra cautella , a species in which current male ejaculate traits suggest previous selection for paternity protection consistent with a sperm-competitive environment. Larvae were reared at high (H) or low population sizes (L). We recorded larval development time, adult longevity and male gametic investment. Our results show a response by adults to signals in the juvenile environment. H males transferred more apyrene, but not eupyrene sperm. We also examined potential trade-offs between somatic characters and reproductive behaviours. Larval duration was longer for H individuals, females and heavier individuals. Further, H females and L males lived longer than L females. Our data are consistent with the theory that males should adjust their reproductive investment in accordance with sperm competition risk.     

Building a Beetle: How Larval Environment Leads to Adult Performance in a Horned Beetle

The link between the expression of the signals used by male animals in contests with the traits which determine success in those contests is poorly understood. This is particularly true in holometabolous insects such as horned beetles where signal expression is determined during metamorphosis and is fixed during adulthood, whereas performance is influenced by post-eclosion feeding. We used path analysis to investigate the relationships between larval and adult nutrition, horn and body size and fitness-related traits such as strength and testes mass in the horned beetle Euoniticellus intermedius. In males weight gain post-eclosion had a central role in determining both testes mass and strength. Weight gain was unaffected by adult nutrition but was strongly correlated with by horn length, itself determined by larval resource availability, indicating strong indirect effects of larval nutrition on the adult beetle’s ability to assimilate food and grow tissues. Female strength was predicted by a simple path diagram where strength was determined by eclosion weight, itself determined by larval nutrition: weight gain post-eclosion was not a predictor of strength in this sex. Based on earlier findings we discuss the insulin-like signalling pathway as a possible mechanism by which larval nutrition could affect adult weight gain and thence traits such as strength.     

Odonates as Indicators of Shallow Lake Restoration by Liming: Comparing Adult and Larval Responses

Odonate assemblages were compared between replicate sets of shallow lakes that had been created and acidified by open‐cast mining across a large area (2,451 ha) of southwest France (Arjuzanx, Landes); one set of lakes (n = 5) was experimentally restored by liming with calcium carbonate, whereas another group (n = 5) was left as untreated reference lakes. Both odonate adults and exuviae were sampled bimonthly during May–August 1998. Elevated turbidity and conductivity in limed lakes were the only physicochemical measures differing between restored and reference lakes, because deacidification occurred naturally, even in reference lakes during the 17 years after the onset of restoration. Restoration by liming can apparently lead to effects on lake turbidity that might be considered adverse. Twenty‐four and 19 odonate species occurred among adults and exuviae, respectively, but there were no significant differences in richness between restored and reference sites. However, significantly, more exuviae were collected from the reference sites (588 vs. 180), where exuvial diversity and rank abundance indicated more evenly structured assemblages than those in restored lakes. Ordination showed that adult assemblages differed significantly between restored and reference lakes, and varied highly significantly with lake turbidity. This effect occurred because a small group of generally scarce adults were characteristic of reference sites (Chalcolestes viridis, Lestes virens, Cordulia aenae, Leucorrhinia albifrons, and Sympetrum sanguineum). Exuviae of these same species were less abundant at restored sites, but exuvial assemblages overall did not discriminate between restored and reference lakes. We conclude that lake restoration by liming can reduce diversity and larval numbers among odonates and subtly affects adult assemblages. In this case study, adult assemblages discriminated best between the lake types involved in the experiment, but important additional information arose from exuvial abundance and structure. This study indicates that natural recovery processes after acidification in formerly open‐cast areas––rather than chemical intervention through liming––might lead to preferable conservation outcomes.     

Requirements for Hedgehog, a Segmental Polarity Gene, in Patterning Larval and Adult Cuticle of Drosophila

Mutations of the hedgehod gene are generally embryonic lethal, resulting in a lawn of denticles on the ventral surface. In strong alleles, no segmentation is obvious and the anteroposterior polarity of ventral denticles is lost. Temperature shift analysis of a temperature-sensitive allele indicates an embryonic activity period for hedgehod between 2.5 and 6 hr of embryonic development (at 25 degrees) and a larval/pupal period from 4 to 7 days of development (at 25 degrees). Mosaic analysis of hedgehod mutations in the adult cuticle indicates a series of defined defects associated with the failure of appropriate hedgehod expression. In particular, defects in the distal portions of the legs and antenna occur in association with homozygous hedgehog clones in the posterior compartment of those structures. Because the defects are associated with homozygous clones, but are not co-extensive, a type of "domineering" nonautonomy is proposed for the activity of the hedgehog gene.     

Long-Term Changes in the Distributions of Larval and Adult Fish in the Northeast U.S. Shelf Ecosystem

Many studies have documented long-term changes in adult marine fish distributions and linked these changes to climate change and multi-decadal climate variability. Most marine fish, however, have complex life histories with morphologically distinct stages, which use different habitats. Shifts in distribution of one stage may affect the connectivity between life stages and thereby impact population processes including spawning and recruitment. Specifically, many marine fish species have a planktonic larval stage, which lasts from weeks to months. We compared the spatial distribution and seasonal occurrence of larval fish in the Northeast U.S. Shelf Ecosystem to test whether spatial and temporal distributions changed between two decades. Two large-scale ichthyoplankton programs sampled using similar methods and spatial domain each decade. Adult distributions from a long-term bottom trawl survey over the same time period and spatial area were also analyzed using the same analytical framework to compare changes in larval and adult distributions between the two decades. Changes in spatial distribution of larvae occurred for 43% of taxa, with shifts predominately northward (i.e., along-shelf). Timing of larval occurrence shifted for 49% of the larval taxa, with shifts evenly split between occurring earlier and later in the season. Where both larvae and adults of the same species were analyzed, 48% exhibited different shifts between larval and adult stages. Overall, these results demonstrate that larval fish distributions are changing in the ecosystem. The spatial changes are largely consistent with expectations from a changing climate. The temporal changes are more complex, indicating we need a better understanding of reproductive timing of fishes in the ecosystem. These changes may impact population productivity through changes in life history connectivity and recruitment, and add to the accumulating evidence for changes in the Northeast U.S. Shelf Ecosystem with potential to impact fisheries and other ecosystem services.     

Performance of Five Food Regimes on Anopheles gambiae Senso Stricto Larval Rearing to Adult Emergence in Insectary

Background

Rearing of Anopheles gambiae s.s mosquitoes in insectary with quality cheap food sources is of paramount importance for better and healthy colony. This study evaluated larval survival and the development rate of aquatic stages of An.gambiae s.s under five food regimes; tetramin fish food (a standard insectary larval food), maize pollen, Cerelac, green filamentous algae and dry powdered filamentous algae.

Methods

Food materials were obtained from different sources, cerelac was made locally, fresh filamentous algae was taken from water bodies, dry filamentous algae was ground to powder after it was dried under shade, and maize pollen was collected from the flowering maize. Each food source type was used to feed three densities of mosquito larvae 20, 60, and 100 in six replicates each. Larval age structure was monitored daily until pupation and subsequently adult emergence. Tetramin was used and taken as a standard food source for An. gambiae s.s. larvae feeding in Insectary.

Results

Larval survivorship using maize pollen and Tetramin fish food was statistically insignificant (P = 0.564). However when compared to other food regime survivorship was significantly different with Tetramin fish food performing better than cerelac (P<0.001), dry algae (P<0.001) and fresh algae (P<0.001). The pupation rates and sex ratio of emerging adults had significant differences among the food regimes.

Conclusion

The findings of this study have shown that maize pollen had closely similar nutritional value for larval survivorship to tetramin fish food, a standard larvae food in insectary. Further studies are required to assess the effect of food sources on various life traits of the emerged adults.     

Effects of Low Salinity on Adult Behavior and Larval Performance in the Intertidal Gastropod Crepipatella peruviana (Calyptraeidae)

Shallow-water coastal areas suffer frequent reductions in salinity due to heavy rains, potentially stressing the organisms found there, particularly the early stages of development (including pelagic larvae). Individual adults and newly hatched larvae of the gastropod Crepipatella peruviana were exposed to different levels of salinity stress (32(control), 25, 20 or 15), to quantify the immediate effects of exposure to low salinities on adult and larval behavior and on the physiological performance of the larvae. For adults we recorded the threshold salinity that initiates brood chamber isolation. For larvae, we measured the impact of reduced salinity on velar surface area, velum activity, swimming velocity, clearance rate (CR), oxygen consumption (OCR), and mortality (LC50); we also documented the impact of salinity discontinuities on the vertical distribution of veliger larvae in the water column. The results indicate that adults will completely isolate themselves from the external environment by clamping firmly against the substrate at salinities ≤24. Moreover, the newly hatched larvae showed increased mortality at lower salinities, while survivors showed decreased velum activity, decreased exposed velum surface area, and decreased mean swimming velocity. The clearance rates and oxygen consumption rates of stressed larvae were significantly lower than those of control individuals. Finally, salinity discontinuities affected the vertical distribution of larvae in the water column. Although adults can protect their embryos from low salinity stress until hatching, salinities <24 clearly affect survival, physiology and behavior in early larval life, which will substantially affect the fitness of the species under declining ambient salinities.