Variables Influencing the Effect of a Meal on Brain Tryptophan

Previous work from our laboratory points to plasma free tryptophan being a useful predictor of brain tryptophan concentration in many circumstances. Other work, in particular various studies on the acute effects of food intake, has emphasized the roles of plasma total tryptophan and of plasma large neutral amino acids that compete with tryptophan for transport to the brain. We have now studied associations between the above variables under different dietary conditions. Rats were allowed to feed for restricted periods during a 12-h light-12-h dark cycle. In the first study, rats were given access to a carbohydrate diet for 2 h midway through the light cycle and following an 18-h fast. The resultant rise of brain tryptophan was explicable largely by the associated fall in large neutral amino acids. In a second study, rats were adapted to a regimen whereby they were allowed access to the standard laboratory diet for 4 h during the dark cycle for 3 weeks. A postprandial decrease in brain tryptophan was associated with a fall in free tryptophan and of its ratio to competing amino acids. The brain change could be attributed neither to changes in plasma total tryptophan (which increased) nor to changes of its ratio to the competers (which remained unchanged). Results as a whole are thus consistent with changes of plasma free tryptophan and large neutral amino acid concentrations affecting brain tryptophan concentration under different dietary circumstances. It is suggested that these influences serve to maintain brain tryptophan when dietary supplies are defective.     

Dynamical systems basis of metamorphosis: diversity and plasticity of cellular states in reaction diffusion network

Dynamics maintaining diversity of cell types in a multi-cellular system are studied in relation to the plasticity of cellular states. By adopting a simple theoretical framework for intra-cellular chemical reaction dynamics and considering the division and death of cells, the development of cells is studied. Cell differentiation process is found to occur through instability in transient dynamics and cell-cell interaction. In long-term behavior, extinction of multiple cells is repeated, which leads to itinerancy over successive quasi-stable multi-cellular states consisting of different types of cells. By defining the plasticity of a cellular state, it is shown that the plasticity of cells decreases before the extinction of most cell types, from which diversity and plasticity are later recovered. In the following, a decrease of plasticity occurs again, leading to the next extinction. This cycle of diversification and extinction is repeated. Relevance of our results to development and evolution is briefly discussed.     

Molecular Cloning of a Preprohormone from Hydra magnipapillata Containing Multiple Copies of Hydra-LWamide (Leu-Trp-NH2) Neuropeptides: Evidence for Processing at Ser and Asn Residues

Abstract: The simple, freshwater polyp Hydra is often used as a model to study development in cnidarians. Recently, a neuropeptide, 2, has been isolated from sea anemones that induces metamorphosis in a hydroid planula larva to become a polyp. Here, we have cloned a preprohormone from Hydra magnipapillata containing 11 (eight different) immature neuropeptide sequences that are structurally related to the metamorphosis-inducing neuropeptide from sea anemones. During the final phase of our cloning experiments, another research team independently isolated and sequenced five of the neuropeptides originally found on the preprohormone. Comparison of these mature neuropeptide structures with the immature neuropeptide sequences on the preprohormone shows that most immature neuropeptide sequences are preceded by Ser or Asn residues, indicating that these residues must be novel processing sites. Thus, the structure of the Hydra prepro-hormone confirms our earlier findings that cnidarian pre-prohormones contain unusual or novel processing sites. Nearly all neuropeptide copies located on the Hydra preprohormone will give rise to mature neuropeptides with a C-terminal Gly-Leu-Trp-NH₂ sequence (the most frequent one being Gly-Pro-Pro-Pro-Gly-Leu-Trp-NH₂; Hydra-LWamide I; three copies). Based on their structural similarities with the metamorphosis-inducing neuropeptide from sea anemones, the mature peptides derived from the Hydra-LWamide preprohormone are potential candidates for being developmentally active neurohormones in Hydra .     

Change in significance of feeding during larval development in the yellow-spotted longicorn beetle, Psacothea hilaris

Larvae of the west-Japan type yellow-spotted longicorn beetle, Psacothea hilaris (Coleoptera: Cerambycidae), show a long-day type photoperiodic response at 25 degrees C; under long-day conditions, larvae pupate after the fourth or fifth instar, while under short-day conditions, they undergo a few nonstationary supernumerary molts and eventually enter diapause. In the present study, the effect of food on the development and photoperiodic response of the larvae was examined with special reference to molting and pupation. Although the pupal body size was greatly affected by the food quality and the length of feeding, the critical day length for induction of metamorphosis at 25 degrees C was always between 13.5 and 14 h. Exposure to starvation of larvae reared on the standard diet revealed that the capability to pupate is acquired after a few days of feeding in the fourth instar. In the larvae that had acquired the capability to pupate, premature pupation was induced by exposure to starvation, indicating that feeding becomes dispensable long before it is normally terminated.     

Growth and nutritional condition of the larvae of Odontesthes microlepidotus (Atherinidae): an experimental approach

Synopsis The relation between the feeding and mortality of fish larvae is mediated not only by starvation but also by their growth. In this work, daily otolith ring deposition was analysed in laboratory-reared larvae of Odontesthes microlepidotus (Jenyns), and their growth was studied in relation to ring number and otolith diameter. The nutritional condition of these larvae was assessed by means of morphometric comparisons between fed and starved larvae in order to develop a tool to evaluate the nutritional status of larvae in breeding areas. Changes in body shape have been found as a consequence of starvation. No dependence was found between mean hepatocyte nuclear area and fasting.     

Modulation of larval nutrition affects midgut neutral lipid storage and temporal pattern of transcription factor expression during mosquito metamorphosis

During holometabolous insect development the critical weight marks a physiological transition after which juvenile hormone (JH) concentration decreases to such a level that a subsequent increase in ecdysone titer will initiate metamorphosis. Starvation experiments indicate that the Aedes aegypti critical weight is achieved by 24 h after the last larval-larval molt. When grown at 24 degrees C with excess food, the time between the critical weight and maximum weight (interval to cessation of growth) is about 24 h and pupation occurs about 24 h after the maximum weight is achieved. Oil Red O staining of 3rd and early 4th instars indicates that the midgut is a neutral lipid storage organ during this period. Coincident with the attainment of the critical weight is the depletion of stored midgut neutral lipid. Application of methoprene to 24 h post-molt 4th instars results in renewed midgut storage of neutral lipid suggesting that midgut neutral lipid storage is a JH dependent process. Starvation of 4th instars during the 24 h post-molt period suspends development in a fraction of the larvae, and with the resumption of feeding, development resumes. A regimen of starvation and resumption of feeding of 4th instars suggests that JH concentration decreased over a 40 h period after the resumption of feeding and maximum weight is attained about 48 h after the resumption of feeding. We hypothesize that this results in a shortening of the interval to cessation of growth. Real-time PCR experiments indicate that shortening the interval to cessation of growth compresses the time period during which increases in AHR3, AaEcR-B and AaUSP-a expression occur.     

Eye movement dynamics and cognitive self-organization in typical and atypical development

This study analyzed distributions of Euclidean displacements in gaze (i.e. “gaze steps”) to evaluate the degree of componential cognitive constraints on audio-visual speech perception tasks. Children performing these tasks exhibited distributions of gaze steps that were closest to power-law or lognormal distributions, suggesting a multiplicatively interactive, flexible, self-organizing cognitive system rather than a component-dominant stipulated cognitive structure. Younger children and children diagnosed with an autism spectrum disorder (ASD) exhibited distributions that were closer to power-law than lognormal, indicating a reduced degree of self-organized structure. The relative goodness of lognormal fit was also a significant predictor of ASD, suggesting that this type of analysis may point towards a promising diagnostic tool. These results lend further support to an interaction-dominant framework that casts cognitive processing and development in terms of self-organization instead of fixed components and show that these analytical methods are sensitive to important developmental and neuropsychological differences.     

Evidence of a hemolymph-born factor that induces onset of maturation in Manduca sexta larvae

Insect metamorphosis is a complex developmental transition determined and coordinated by hormonal signaling that begins at a critical weight late in the larval phase of life. Even though this hormonal signaling is well understood in insects, the internal factors that are assessed at the critical weight and that drive commitment to metamorphosis have remained unresolved in most species. The critical weight may represent either an autonomous decision by the neuroendocrine system without input from other developing larval tissues, or an assessment of developmental thresholds occurring throughout the body that are then integrated by the neuroendocrine tissues. The latter hypothesis predicts that there could be one or more developmental threshold signals that originate from developing tissues and ultimately induce the onset of metamorphosis. However, there is no evidence for such a signal in the organisms for which the critical weight is well described. Here we test for the evidence of this factor in Manduca sexta (Lepidoptera: Sphingidae) by transferring hemolymph from individuals that are either post- or pre-critical weight into pre-critical weight 5^th instar larvae. We found that hemolymph from a post-critical weight donor induces a shortening of development time, though the mass at pupation is unaffected. This suggests that metamorphic commitment occurring at the critical weight is at least partially coordinated by signaling from developing tissues via a hemolymph-borne signaling factor.     

Analysis of molecular markers for metamorphic competency and their response to starvation or feeding in the mosquito, Aedes aegypti (Diptera: Culicidae)

The nutritional condition of fourth instar larvae of the yellow fever mosquito, Aedes aegypti, governs female longevity and egg production, both are key determinants of pathogen transmission. As well, nutrition provisions larval growth and development and attains its greatest pace in the last larval instar in preparation for metamorphosis to an adult. These developmental processes are regulated by a complex endocrine interplay of juvenile hormone, neuropeptides, and ecdysteroids that is nutrition sensitive. We previously determined that feeding for only 24 h post-ecdysis was sufficient for fourth instar Ae. aegypti larvae to reach critical weight and accumulate sufficient nutritional stores to commit to metamorphosis. To understand the genetic basis of metamorphic commitment in Ae. aegypti, we profiled the expression of 16 genes known to be involved in the endocrine and nutritional regulation of insect metamorphosis in two ways. The first set is a developmental profile from the beginning of the fourth instar to early pupae, and the second set is for fourth instars starved or fed for up to 36 h. By comparing the two sets, we found that seven of the genes (AaegCYP302, AaegJHE43357, AaegBrCZ4, AaegCPF1-2, AaegCPR-7, AaegPpl, and AaegSlif) were expressed during metamorphic commitment in fourth instars and in fed but not starved larvae. Based on these results, the seven genes alone or in combination may serve as molecular indicators of nutritional and metamorphic status of fourth instar Ae. aegypti larvae and possibly other mosquito species in field and laboratory studies to gauge sub-lethal effects of novel and traditional cultural or chemical controls.     

Sublethal Effects of Nitrite on Eastern Tiger Salamander (<Emphasis Type=Italic>Ambystoma tigrinum tigrinum</Emphasis>) and Wood Frog (<Emphasis Type=Italic>Rana sylvatica</Emphasis>) Embryos and Larvae: Implications for Field Populations

Ephemeral pools, which can have high animal biomass and low dissolved oxygen, may be prone to nitrite accumulation. As such, it is important to understand how exposure to nitrite might affect development and growth of amphibians that breed in these ephemeral pools. Wood frog (Rana sylvatica) and eastern tiger salamander (Ambystoma tigrinum tigrinum) embryos and tadpoles and young larvae were exposed to elevated concentrations of nitrite derived from sodium nitrite: 0, 0.3, 0.6, 1.2, 2.1, 4.6, and 6.1 mg l⁻¹ NO₂–N. Increasing nitrite exposure slowed embryonic and larval development in both the eastern tiger salamander and the wood frog, reduced growth in tiger salamander embryos and larvae, and delayed metamorphosis in the wood frog. At concentrations less than 2 mg l⁻¹ NO₂–N nitrite delayed hatching, and at concentrations above 2 mg l⁻¹ time to hatching decreased causing more individuals to hatch at less developed stages. Nitrite also increased asynchrony in tiger salamander hatching. The sublethal effects of nitrite on amphibian development, growth and hatching could have serious repercussions on amphibian fitness in ephemeral environments. Potential increases in mortality on field populations caused by sublethal effects of nitrite are discussed.     

Cloning of a novel murine gene Sfmbt, Scm-related gene containing four mbt domains, structurally belonging to the Polycomb group of genes

We cloned six cDNAs by screening cDNA libraries of cypris larvae from barnacles, Balanus amphitrite, and studied their expression by Northern blot analysis. All of them are expressed in the cypris larvae at the settlement stage, but not in the earlier nauplii larvae nor in later adult barnacles. Therefore, we designated them as barnacle cypris larva-specific genes (bcs); bcs-1, bcs-2, bcs-3, bcs-4, bcs-5 and bcs-6. During the process of larval attachment and metamorphosis, the amounts of bcs-1 and bcs-2 mRNAs decreased, whereas the bcs-3, bcs-4, bcs-5 and bcs-6 mRNAs increased. A homology search showed that all cDNAs encode novel peptides containing characteristic amino acid sequences. This study strongly suggests that these bcs gene products are involved in the cypris larval attachment and metamorphosis of barnacles.     

The functions of polarized water and membrane lipids: a rebuttal

In 1973, experimental evidence was reported in support of the view that water polarized in multilayers rather than simple lipid is responsible for the semipermeable properties of living cell membranes. The present article presents a full rebuttal to a subsequent attack on that view.     

Short-term fluctuation in salinity promotes rapid larval development and metamorphosis in Dendraster excentricus

The effect of constant and fluctuating salinity on larval development and metamorphosis of the sand dollar Dendraster excentricus was investigated in the laboratory. Sand dollar larvae at different stages of development were kept either at 32‰ (controls), exposed to constant low salinity (22‰) throughout development, or exposed to fluctuating salinity (i.e. transferring larvae from 32‰ to 22‰ for 7 days then back to 32‰ for the rest of their development). Larvae exposed to constant low salinity were significantly smaller but developed all larval arms at a slower rate than larvae in all other treatments. Larvae exposed to fluctuating salinity recovered and developed significantly longer larval arms and bigger rudiments than larvae kept at constant low salinity. Larvae exposed to fluctuating salinity produced more juveniles than larvae at constant high salinity (32‰), while those at constant low salinity produced few or no juveniles. Four-arm larvae exposed to fluctuating salinity produced significantly more juveniles than six-arm larvae exposed to the same treatment. Transferring competent 8-arm larvae from 31‰ to 15‰ for 2 days then back to 31‰, induced metamorphosis with juvenile production being significantly higher than for those kept at a constant salinity of 20, 25 and 31‰. This study indicates that a short-term decrease in salinity might induce metamorphosis for this species.     

Pattern of self-organization in tumour systems: complex growth dynamics in a novel brain tumour spheroid model

We propose that a highly malignant brain tumour is an opportunistic, self-organizing and adaptive complex dynamic biosystem rather than an unorganized cell mass. To test the hypothesis of related key behaviour such as cell proliferation and invasion, we have developed a new in vitro assay capable of displaying several of the dynamic features of this multiparameter system in the same experimental setting. This assay investigates the development of multicellular U87MGmEGFR spheroids in a specific extracellular matrix gel over time. The results show that key features such as volumetric growth and cell invasion can be analysed in the same setting over 144 h without continuously supplementing additional nutrition. Moreover, tumour proliferation and invasion are closely correlated and both key features establish a distinct ratio over time to achieve maximum cell velocity and to maintain the system's temporo-spatial expansion dynamics. Single cell invasion follows a chain-like pattern leading to the new concept of a intrabranch homotype attraction . Since preliminary studies demonstrate that heterotype attraction can specifically direct and accelerate the emerging invasive network, we further introduce the concept of least resistance, most permission and highest attraction as an essential principle for tumour invasion. Together, these results support the hypothesis of a self-organizing adaptive biosystem.     

A quantitative dynamical systems approach to differential learning: self-organization principle and order parameter equations

Differential learning is a learning concept that assists subjects to find individual optimal performance patterns for given complex motor skills. To this end, training is provided in terms of noisy training sessions that feature a large variety of between-exercises differences. In several previous experimental studies it has been shown that performance improvement due to differential learning is higher than due to traditional learning and performance improvement due to differential learning occurs even during post-training periods. In this study we develop a quantitative dynamical systems approach to differential learning. Accordingly, differential learning is regarded as a self-organized process that results in the emergence of subject- and context-dependent attractors. These attractors emerge due to noise-induced bifurcations involving order parameters in terms of learning rates. In contrast, traditional learning is regarded as an externally driven process that results in the emergence of environmentally specified attractors. Performance improvement during post-training periods is explained as an hysteresis effect. An order parameter equation for differential learning involving a fourth-order polynomial potential is discussed explicitly. New predictions concerning the relationship between traditional and differential learning are derived.     

Spiny lobster development: mechanisms inducing metamorphosis to the puerulus: a review

This review outlines current knowledge of mechanisms effecting metamorphosis in decapod crustaceans and insects. The comparative approach demonstrates some of the complexities that need resolving to find an answer to the question raised frequently by ecologists: “What triggers metamorphosis in spiny lobsters?” It is evident that crustacean moulting and metamorphosis are genetically controlled through endocrine systems that mediate gene expression. The molecular mechanisms underlying these developmental processes have been studied intensively in insects, particularly in the fruitfly, Drosophila melanogaster (Diptera), and some lepidopteran species. Comparatively, there is minimal information available for a few decapod crustacean species, but none for spiny lobsters (Palinuridae). Nothing was known of hormone signalling transduction pathways, via nuclear receptors (NRs) and gene activation during larval moults in palinurids—until a recent, ground-breaking study of early phyllosomal development of Panulirus ornatus by Wilson et al. (Rock Lobster Enhancement and Aquaculture Subprogram. FRDC Project 2000/263, Australian Govt, Fisheries Research and Development Corporation and Australian Institute of Marine Science, Nov 2005). Their study not only identified homologues of five hormone NRs of D. melanogaster, but also patterns of gene regulation showing strong similarities to those of gene expression found in insect larval development. Their results indicated that control of moulting and metamorphosis in palinurids closely parallels that in insects, suggesting that insects can serve as model systems for elucidating molecular mechanisms in larval decapods. In insects and crustaceans, the steroid hormone, ecdysone, (20E) initiates moulting. In insects, juvenile hormone (JH) mediates the type of larval moult that occurs, either anamorphic or metamorphic. The latter results when the level of JH in the haemolymph drops in the final larval instar. High levels of JH inhibit the metamorphic moult during insect larval development. The interaction of 20E and JH is not fully understood, and the operative molecular mechanisms are still being elucidated. No nuclear receptor for JH has been identified, and alternative JH signalling pathways await identification. In decapod crustaceans, methyl farnesoate (MF), a precursor of JH, replaces the latter in other functions mediated by JH in insects; but there is little evidence indicating that MF plays a similar ‘antimetamorphic’ role in decapod larval moults.