The Molecular Ecology of Fundulus heteroclitus Hemoglobin-Oxygen Affinity

Animals faced with environmental perturbations must adapt orface extinction. The respiratory complex, specifically hemoglobins,is perhaps the best system to study such adaptation becauseit exists at the organism—environment interface. Fishare particularly useful models because they respond directlyto such environmental variables as temperature, oxygen, pH,carbon dioxide, and salinity. Our experiments have addressedthe molecular, cellular, and physiological mechanisms employedby fish to maintain respiratory homeostasis in the wake of changingtemperature and oxygen. Immediate, intermediate, and long-termadaptation can only be understood when the hemoglobin's ligandbinding properties and the cellular and hormonal regulationof various ligands are considered simultaneously. We describea detailed thermodynamic model for the binding of oxygen, protons,and organic phosphates to hemoglobin; discuss the role of multiplehemoglobins; and present evidence for physiological and geneticregulation of hemoglobin's major allosteric modifiers in responseto environmental stress in the mummichog, Fundulus heteroclitus     

Cold-Tolerant Crop Species Have Greater Temperature Homeostasis of Leaf Respiration and Photosynthesis Than Cold-Sensitive Species

Some plant species show constant rates of respiration and photosynthesismeasured at their respective growth temperatures (temperaturehomeostasis), whereas others do not. However, it is unclearwhat species show such temperature homeostasis and what factorsaffect the temperature homeostasis. To analyze the inherentability of plants to acclimate respiration and photosynthesisto different growth temperatures, we examined 11 herbace-ouscrops with different cold tolerance. Leaf respiration (R_area)and photosynthetic rate (P_area) under high light at 360 µll^–1 CO₂ concentrations were measured in plants grown at15 and 30°C. Cold-tolerant species showed a greater extentof temperature homeostasis of both R_area and P_area than cold-sensitivespecies. The underlying mechanisms which caused differencesin the extent of temperature homeostasis were examined. Theextent of temperature homeostasis of P_area was not determinedby differences in leaf mass and nitrogen content per leaf area,but by differences in photosynthetic nitrogen use efficiency(PNUE). Moreover, differences in PNUE were due to differencesin the maximum catalytic rate of Rubisco, Rubisco contents andamounts of nitrogen invested in Rubisco. These findings indicatedthat the temperature homeostasis of photosynthesis was regulatedby various parameters. On the other hand, the extent of temperaturehomeostasis of R_area was unrelated to the maximum activity ofthe respiratory enzyme (NAD-malic enzyme). The R_area/P_area ratiowas maintained irrespective of the growth temperatures in allthe species, suggesting that the extent of temperature homeostasisof R_area interacted with the photosynthetic rate and/or thehomeostasis of photosynthesis.     

华硬蜱和二棘血蜱的交叉免疫反应

本文首次比较了经中华硬蜱(Ixodes sinensis)叮咬三次后再经二棘血蜱(Haimaphysalis bispinosa)叮咬的家兔与仅经二棘血蜱叮咬的家兔的交叉免疫抗性。二棘血蜱叮咬被中华硬蜱致敏的家兔时,吸血增重为：143.12±32.67mg,但二棘血蜱在正常家兔体上寄生,初次吸血增重为：181.30±44.35mg,两者之间有显著性差异(P<0.01)。中华硬蜱和二棘血蜱唾液腺提取物(SGE)经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),显示两者分别有24条和22条电泳带,中华硬蜱主带有6条,分子量分别为142、105、94、66/65、64和56kD,而二棘血蜱主带有5条,分子量分别为：215、114、105、66/65和58kn,经中华硬蜱叮咬致敏的家兔血清和经二棘血蜱叫‘咬致敏的家兔血清作免疫印渍,均显示出105kD这一电泳带。该实验表明中华硬蜱和二棘血蜱叮咬家兔两者之间存在着交叉免疫反应,提示105kD蛋白质抗原可能是两者的共同抗原。     

Physiological epistasis, ontogenetic conflict and natural selection on physiology and life history

Ontogenetic conflict arises when optima for alleles governingfitness variation differ between juveniles and adults or betweenadult sexes. Loci that govern development of alternative phenotypesin the sexes, hereafter termed morph-determining loci, mediatedevelopment through the endocrine system. Morphotypic selectionis defined to be multivariate selection favoring discrete alternativemorphotypes (e.g., optima). When the optimal combinations ofalleles for alternative morphs differ between the sexes, itgenerates conflicting selection pressure and thus ontogeneticconflict. Selection on morph alleles promotes ontogenetic conflictbecause it perturbs physiological epistasis that governs theexpression of male versus female traits. Expression of physiologicaltraits arises from homeostasis that maintains trait expressionwithin a normal range. The genetic basis of homeostasis is likelyto arise from interactions among several genes (e.g., geneticepistasis) or protein products (e.g., physiological epistasis).For example, endocrine regulation arises from interactions betweengondatropins, which are protein hormones produced by the hypothalamic-pituitaryglands, and steroid hormones, which are produced by the gonads(e.g., HPG axis). The side-blotched lizard system is discussedwith respect to physiological bases of ontogenetic conflict.We also describe a novel molecular marker strategy for uncoveringgenome-wide physiological epistasis in nature. Finally, ontogeneticconflict exerts selection on females to evolve mate selectionor cryptic choice that is reflected in different sires beingchosen for son versus daughter production. We describe how side-blotchedlizard females ameliorate ontogenetic conflict by cryptic choiceof male genotypes to produce sons versus daughters.     

Efficiency as a foraging currency in animals attaining a gain below the energetic ceiling

Previous research has found that efficiency, or, more precisely,the foraging gain ratio (FGR), is a valid currency in foragingtheory when (1) there is a limit to the energy that can beassimilated by the forager and (2) a forager is trying to meetan energy requirement. The FGR is b/ (c — c_r), whereb is the rate of metabolizable energy intake, and c and c_rare the rates of energy expenditure while foraging and resting,respectively. Here I show that, when energy expenditure hasa cost besides energy, animals should also choose the optionwith the highest FGR when they are aiming at a given positivedaily gain. The next question is which gain they should aimfor? Researchers have shown that observed intake levels ofgrowing ruminants are close to the levels predicted by maximizationof the efficiency of oxygen utilization. This currency can be approximated by (B — C + C_r) / C, where B is the daily metabolizable energy intake, and C and C_r are the total andbasal daily energy expenditures, respectively. By simulatinggrowth at different intake levels, I found that mass-specificoxygen consumption rate is indeed minimal at the observed intakelevels. This is the first study in which these efficiency measures(FGR and the efficiency of oxygen utilization) are combined.     

Kingdom animalia: The zoological malaise from a microbial perspective

Pain and cognitive dissonance abounds amongst biologists: theplant-animal, botany-zoology wound has nearly healed and thenew gash—revealed by department and budget reorganizations—is"molecular" vs. "organismic" biology. Here I contend that resolutionof these tensions within zoology requires that an autopoietic-gaianview replace a mechanical-neodarwinian perspective; in the interestof brevity and since many points have been discussed elsewhere,rather than develop detailed arguments I must make staccatostatements and refer to a burgeoning literature. The first centralconcept is that animals, all organisms developing from blastularembryos, evolved from single protist cells that were unableto reproduce their undulipodia. The second points to the usefulnessof recognizing the analogy between cyclically established symbiosesand meiotic sexuality     

Ethogram of Abdopus aculeatus (d'Orbigny, 1834) (Cephalopoda: Octopodidae): Can behavioural characters inform octopodid taxomony and systematics?

An ethogram is provided for the small, intertidal, diurnal octopusAbdopus aculeatus (d'Orbigny, 1834). Information is based primarilyon in situ observations of adults, and supplemented with photographsof animals in aquaria. Aspects of the dymantic display, matingsystem, activity patterns, and habitat use appear similar tothose expressed by other members of Abdopus, as well as thelarge sister taxon Octopus cyanea, suggesting that these behavioursmay be conserved throughout the evolution of these octopuses.Many skin components are also shared with Octopus bimaculoides,which may reflect either an evolutionary affinity with thisoctopus, or convergence in these characters. If behaviouralunits such as those documented here are compiled in a consistentmanner for other species, then they may facilitate taxonomicidentification, as well as future evolutionary studies of octopodids.     

Evolution of the Toxoglossa venom apparatus as inferred by molecular phylogeny of the Terebridae

Toxoglossate marine gastropods, traditionally assigned to thefamilies Conidae, Terebridae, and Turridae, are one of the mostpopulous animal groups that use venom to capture their prey.These marine animals are generally characterized by a venomapparatus that consists of a muscular venom bulb and a tubularvenom gland. The toxoglossan radula, often compared with a hypodermicneedle for its use as a conduit to inject toxins into prey,is considered a major anatomical breakthrough that assistedin the successful initial radiation of these animals in theCretaceous and early Tertiary. The pharmacological success oftoxins from cone snails has made this group a star among biochemistsand neuroscientists, but very little is known about toxins fromthe other Toxoglossa, and the phylogeny of these families islargely in doubt. Here we report the first molecular phylogenyfor the Terebridae and use the results to infer the evolutionof the venom apparatus for this group. Our findings indicatethat most of the genera of terebrids are polyphyletic, and onespecies ("Terebra" (s.l.) jungi) is the sister group to allother terebrids. Molecular analyses combined with mapping ofvenom apparatus morphology indicate that the Terebridae havelost the venom apparatus at least twice during their evolution.Species in the genera Terebra and Hastula have the typical venomapparatus found in most toxoglossate gastropods, but all otherterebrid species do not. For venomous organisms, the dual analysisof molecular phylogeny and toxin function is an instructivecombination for unraveling the larger questions of phylogenyand speciation. The results presented here suggest a paradigmshift in the current understanding of terebrid evolution, whilepresenting a road map for discovering novel terebrid toxins,a largely unexplored resource for biomedical research and potentialtherapeutic drug development.     

Endocrine mediation of vertebrate male alternative reproductive tactics: the next generation of studies

In many species of animals, males may achieve reproductive successvia one of several alternative reproductive tactics. Over thepast decade or so, there has been a concerted effort to investigateendocrine mechanisms that underlie such discrete behavioral(and often morphological) variation. In vertebrates, the firstgeneration of studies focused on potential organizational oractivational effects of steroid hormones (Moore, 1991; Mooreet al., 1998). Some of these studies have made it clear that,in addition to circulating hormone levels, one must also considerother aspects of the endocrine system, including hormone receptors,binding globulins and potential interactions among endocrineaxes. In this paper, I review recent work on endocrine mechanismsand suggest possibilities for future investigation. I highlighthow individual variation in sensitivity to environmental conditions,particularly with respect to various stressors, may accountfor the existence of alternative male reproductive phenotypes.Along these lines, I briefly explain the logic behind our workwith male phenotypes of longear sunfish (Lepomis megalotis)that is aimed at determining the tissue-specific distributionand activity of two enzymes that are common to androgen andglucocorticoid metabolism. A major goal of our work is to examinethe potential role of steroidogenic enzymes in the transductionof environmental information to influence the expression ofalternative male reproductive phenotypes.     

PHYLOGENETIC RELATIONSHIPS OF IBERUS GUALTIERANUS AND I. ALONENSIS (GASTROPODA: HELICIDAE) BASED ON PARTIAL MITOCHONDRIAL 16S rRNA AND COI GENE SEQUENCES

There is controversy about the phylogenetic relationships betweenIberus gualtieranus and I. alonensis. Some authors considerthem as valid species or subspecies while others believe thatthe flattened shell of I. gualtieranus is an ecotypic adaptationto dry karstic environments. Two fragments of the mitochondrialDNA (partial COI and 16S rRNA) were sequenced and used in maximumparsimony, maximum likelihood and neighbour-joining analyses.Iberus alonensis show two distinct lineages, one from Almeríaand the other one from Granada and Jaén-Córdobaregions. Iberus gualtieranus populations are recovered as aterminal node within the I. alonensis group from Almería.The I. gualtieranus clade shows a polytomy and there are nodifferences between the populations of the three isolated localitieswhere I. gualtieranus is currently distributed. This indicatesthat the geographical isolation of these populations has notresulted in genetic diversification. The results indicate thatthe population of I. gualtieranus from Sierra de Gádorin Almería is the only autochthonous one, while the othertwo populations originated by historical introductions. On thebasis of the differences in shell morphology, together withthe presence of a hybrid zone connecting both taxa in nature,and the possibility of obtaining fertile hybrids under laboratoryconditions, we conclude that these two taxa represent two subspecies:Iberus gualtieranus gualtieranus and I. gualtieranus alonensis. (Received 27 September 2004; accepted 10 March 2005)     

NOTES ON THE BEHAVIOR OF THE HYDROID, CORYMORPHA PALMA

Corymorpha, like Porpita, exhibits a synchronous oral flexion,or "concert," of its proximal tentacles. As this is unlike thecontractile writhings of siphonophoran zooids, the concert maybe a significant character in linking the behavior of Porpitamore closely to hydroids than to siphonophores. Except for thisresponse the behavior of Corymorpha and Porpita are different.In Porpita a through-conducting system forms the only link betweententacles, whereas in Corymorpha both local and through-conductingsystems connect tentacles. Corymorpha engages in rhythmic bottom-feedingusing distal tentacles which have no homologue in Porpita. Themechanical properties of the musculo-skeletal system are differentin the two, giving tentacle motions in Corymorpha a continuousappearance, and in Porpita a spasmodic appearance. Reduced glutathione, glycine, DL-serine, L-cysteine and L-glutamicacid can elicit a feeding response in Corymorpha. Sodium thioglycollateand sucrose elicit no response. Glycine is the most effectivestimulant, ca. 5 X 10^–7 M solution being effective. L-glutamicacid gives the weakest response. The presence of chemicals alonecan be sufficient to elicit the feeding reaction in starvedanimals. In moderately well-fed animals, touch may be required.In fully fed animals both touch and chemicals may be insufficient. Parker reports that when Corymorpha is shocked electricallyon its stalk it will bend the stalk so as to apply the hydranthaccurately to the stimulated spot. The response I found wasa curling of the stalk toward the side stimulated, but not toa specific spot on the stalk. Chemical or tactile stimuli maybe necessary for final location of the stimulated site. Parkerreported that the response remained accurate even after severalcuts had been made in the stalk. I could find no evidence forthis.     

家蚕细胞周期蛋白基因BmCcnl1的克隆及表达分析

家蚕Bombyx mori由受精卵到完成胚胎发育孵化的过程中, 细胞进行大量的分裂和分化, 然而滞育性卵的胚胎细胞分化至G2期便停滞在此阶段。为了探索这一发育阶段细胞内的分子调控, 本研究以人Homo sapiens的细胞周期蛋白基因cyclin L1为模板, 成功克隆了家蚕同源基因BmCcnl1（GenBank登录号: FJ889988）。BmCcnl1基因开放阅读框（open reading frame, ORF）全长1 254 bp, 编码417个氨基酸。利用Protean软件分析得出BmCcnl1蛋白预测分子量为49 kDa, 等电点为9.84。利用DNA重组技术构建了BmCcnl1基因的重组表达载体pET-21d-BmCcnl1, 对其进行原核表达, 其表达的蛋白以包涵体形式存在。利用RT-PCR技术分析了BmCcnl1基因在胚胎发育过程中的转录水平, BmCcnl1基因在非滞育性卵的胚胎发育阶段基本保持相对稳定的转录表达, 而滞育性卵从蛾体产下经过72 h后已经检测不到BmCcnl1基因的转录。结果提示, BmCcnl1基因与胚胎期滞育及非滞育性卵的发育调控相关。对该基因的克隆和表达分析为今后研究家蚕胚胎发育及细胞周期调控奠定了基础。     

Genomics of Basal metazoans

An in-depth understanding of the biology of animals will requirethe generation of genomics resources from organisms from allphyla in the metazoan phylogenetic tree. Such resources willideally include complete genome sequences and comprehensiveEST (expressed sequence tag) datasets for each species of interest.Of particular interest in this regard are animals in the earlydiverging non-bilaterian phyla Porifera, Placozoa, Cnidaria,and Ctenophora. Publications describing the results from theuse of genomics approaches in these phyla have only recentlybegun to appear (Kortschak et al., 2003; Yang et al., 2003;Steele et al., 2004). Issues to be considered here include choosingthe basal metazoan species to examine with genomics approaches,the relative advantages and disadvantages of genome sequencingversus EST projects, and the resources and infrastructure requiredto carry out such projects successfully.     

黄粉虫β-1,3-葡聚糖识别蛋白的分离纯化及部分生物学功能

采用中性盐沉淀、凝胶层析等常规方法纯化黄粉虫Tenebrio molitor血淋巴中的β-1,3-葡聚糖识别蛋白,并对其在酚氧化酶原激活系统中的作用进行了初步的研究。结果表明:黄粉虫血淋巴的β-1,3-葡聚糖识别蛋白的分子量约为70 kDa,主要分布于血浆中。纯化的β-1,3-葡聚糖识别蛋白只能特异性地识别β-1,3-葡聚糖而不能识别肽聚糖。在β-1,3- 葡聚糖所诱导的酚氧化酶原的激活过程中,随着酚氧化酶原激活程度的提高,内源性β-1,3-葡聚糖识别蛋白的含量逐渐减少。抗β-1,3-葡聚糖识别蛋白多克隆抗体对黄粉虫血淋巴中由β-1,3-葡聚糖所诱导的酚氧化酶活性起抑制作用,且该抑制作用呈现一种剂量依赖性的趋势。上述结果有助于深入了解β-1,3-葡聚糖对黄粉虫血淋巴酚氧化酶原激活系统的激活作用。     

Aquaporins in complex tissues. I.Developmental patterns in respiratory and glandular tissues of rat

Developmentalexpression of aquaporin water transport proteins is not well understoodin respiratory tract or secretory glands; here we define aquaporinprotein ontogeny in rat. Expression of aquaporin-3 (AQP3), AQP4, andAQP5 proteins occurs within 2 wk after birth, whereas AQP1 firstappears before birth. In most tissues, aquaporin protein expressionincreases progressively, although transient high-level expression isnoted in distal lung (AQP4 at postnatal day+2) and trachea (AQP5 at postnatalday +21 and AQP3 at postnatal day+42). In mature animals, AQP5 is abundant in distallung and salivary glands, AQP3 and AQP4 are present in trachea, andAQP1 is present in all of these tissues except salivary glands.Surprisingly, all four aquaporin proteins are highly abundant innasopharynx. Unlike AQP1, corticosteroids did not induce expression ofAQP3, AQP4, or AQP5 in lung. Our results seemingly implicate aquaporinsin proximal airway humidification, glandular secretion, and perinatalclearance of fluid from distal airways. However, the studies underscorea need for detailed immunohistochemical characterizations anddefinitive functional studies.

     

Morphological and Molecular Resolution of a Putative Cryptic Species Complex: a Case Study of Notoacmea Fascicularis (Menke, 1851) (Gastropoda: Patellogastropoda)

Evidence that Notoacmea fascicularis (Menke, 1851) is a complexof at least two distinct taxa of species rank is ambiguous.A discriminant function analysis of conchological data showsa weak geographic effect, while radular morphology clearly delineatestwo sympatric groups with rare intermediates. Lastly, moleculardata (mt cytochrome c oxidase subunit I) suggests a single speciesand a geographic effect. We consider N. fascicularis to be a singletaxon, variable for radular lateral tooth morphology. In thepast these two different radular morphologies would be indicativeof generic rank. Our knowledge of the intraspecific variability ofmost gastropod characters is poor, and this makes specific identificationsor groupings based on single character systems such as the radulaprecarious. Adequate sampling and evaluation of population-levelcharacter states (conchological, anatomical and molecular) isneeded to identify as well as falsify cryptic species complexes. (Received 28 August 1997; accepted 6 May 1998)     

Lipid-ovary indices in food-limited Daphnia

The amount of lipid carried by a zooplankton individual is highlyvariable. Various lines of evidence have suggested that thenutritional condition of animals could be ascertained via thedetermination of lipid content. However, few studies have beenconducted where the lipid levels of animals of known conditionhave been measured. Here, we report on the lipid index and combinedlipid-ovary index values of Daphnia obrusa reared on foods madeup of varying quantity and quality of a unialgal (Scenedesmusacutus) diet. Lipid levels differed in different food types.However, in no cases were these indices positively correlatedto growth rate, such as would be necessary if one were to usethem to diagnose food limitation in the field. Thus, we urgecaution in the utilization of either the lipid or lipid-ovaryindex until more is known about the role of fatty acids in thediets of these animals.     

Exposure of the Cyanobacterium Synechocystis PCC6803 to Salt Stress Induces Concerted Changes in Respiration and Photosynthesis

In order to survive and to grow in the presence of a high salinity(550 mM NaCl) Synechocystis PCC6803 increases its energeticcapacity. The salt-induced increase of electron transport ratesinvolves both cytochrome c oxidase and photosystem I. In contrast,electron transport rates measured through complexes I plus IIIof the respiratory chain, or through the photosystem II pluscytochrome b₆f complexes of the photosynthetic chain, do notshow appreciable changes. The time at which changes in electrontransport rates occur in the photosystem I and cytochrome coxidase complexes after the onset of salt stress indicates similaritiesin the adaptation of dark respiration and (cyclic) photosyntheticelectron flow. Given an increase of whole cell respiration andof PSI cyclic electron flow larger than the neosynthesis ofcytochrome aa3 and PSI reaction centers would predict, it appearsthat both adaptations require more than just synthesis of thesetwo complexes. (Received April 12, 1993; Accepted August 10, 1993)