Génétique et évolution des Nématodes libres. Perspectives tirées de l'étude deCaenorhabditis elegans

Summary Free-living nematodes are endowed with a set of properties that make them specially attractive for experimental work. Geneticists and evolutionists may be especially interested in their easy cultivation, great fecundity and polymorphism. As an example, some results are brought concerningCaenorhabditis elegans and related species: (1) Sex determination occurs as the expression of a genic balance involving differences in the number of chromosomes. The alternatives gynogenesis, amphimixis, or auto-heterofecundation take a prominent place in the reproductive processes of these species. (2) Polyploidy may be obtained by heat-shock, giving rise to individuals showing characteristic chromosomal unbalance and variability. (3)C. elegans, which cannot normally be raised at a higher temperature than 22°C, has been gradually adapted to temperatures up to 24.5°C. This acclimatization implies an adaptive transformation of the ovarian physiology; this effect is obtained after long training, taking more than 1000 generations. Study of the process of acclimatization shows that its genetical basis may be partly of a non-genic nature, partly of a genic type. Hypotheses are developed for explaining this phenomenon and for the understanding of the evolution from free-living to parasitic nematodes.     

Juvenile hormone and reproduction in the cricket. II. Effect of rearing temperature on corpus allatum activity (in vitro) in adult females

Summary In the Mediterranean field cricket,Gryllus bimaculatus, reproduction is controlled by temperature and the corpus allatum (CA) hormone JH III. In CA of females reared at 24°12°C(168 h) (high reproduction rate) a first peak in JH III synthesis is reached about 4 days earlier than in those of 20°C females (low reproduction rate). Furthermore, in 20°C animals CA activity is low during the entire oviposition period, whereas at 24°12°C high CA activity is found during this period of adult life. The results indicate a stimulation of CA activity and reproduction by thermoperiods around a constant low temperature.Supported by the DFG (SFB 87 A 4).     

Diffusion loading conditions determine recovery of protein synthesis in electroporated P3X63 Ag8 cells

Summary Using the suspension cell line P3X63 Ag8 we have studied the impact of the composition of the diffusion medium on cellular protein synthesis under standard electroporation conditions in TBS-Na. This buffer contains the high saline concentration usually present in electroporation-mediated DNA transfection. Electroporation in the presence of TBS-Na resulted in an immediate shut-off of protein synthesis, even though both FITC-dextran (M_r 40 kD) and Semliki Forest virus core protein (M_r 33 kD) were incorporated efficiently into the cytoplasm across the electropores at 0°C. Subsequent resealing of the pores was completed after a 5-min incubation at 37°C. When compared with control cells, overall protein synthesis of electroporated cells recovered slowly to resume a 30% activity after 1 h of incubation at 37°C. We have determined optimal conditions for diffusion loading (which necessitates the presence of ATP, GTP, amino acids, K⁺, Mg²⁺, and Ca²⁺) and resealing (in the presence of K⁺, Mg²⁺, and Ca²⁺), leading to a full and lasting recovery of protein synthesis within 5 min after pore closure.     

Effect of water temperature on the motility ofPelagia noctiluca (Forksal)

Summary Laboratory tests have been performed, on the behavior of the jellyfishPelagia noctiluca as a function of the water temperature. It has been found that the usual contractions of the umbrella are almost completely missing at 6°C; they begin to appear at about 7–8°C and they reach frequencies of about 10 and 40 per min at 11 and 15°C respectively. An ambient temperature of about 11°C appears to be a threshold value below which this kind of medusa ceases to move actively and sinks, while at higher temperatures it gradually begins to shift, showing a positive thermotropism in the presence of temperature gradients greater than about 0.01 °C/cm.This work was supported by grant No. 80.00748.88 from the National Research Coucil of Italy (CNR).     

Photoperiod-induced changes in temperature-metabolism curve,shivering threshold and body temperature in the pigeon

Summary A short photphase combined with cold-acclimation changed the shivering threshold of the pigeon from 21 to 14 °C. The change of critical temperature was also found in the temperature-metabolism curve. A long photophase both alone and combined with warm-acclimation induced a reduction of body temperature to a lower level (about 2°C) at all ambient temperatures, ranging from 5 40°C.     

Effect of heat treatment on the ATPase activity of various sarcoplasmic reticulum preparations

Summary Ca²⁺-stimulated ATPase activity of sarcoplasmic reticulum (SR) preparations is activated after a short period of preincubation at temperatures between 40 and 45°C, but for temperatures higher than 48°C pronounced denaturation is observed. Heat denaturation is decreased if Mg²⁺ or K⁺ are present during heat treatment.Acknowledgments. This work was supported by research grants from Instituto de Alta Cultura (Grant No. CB/2) and the Calouste Gulbenkian Foundation. We are grateful to Drs.A. P. Carvalho andV. M. C. Madeira for many discussions and suggestions during the course of this work.     

Temperature-controlled alternative respiration and outgrowth rate from conidia ofNeurospora crassa

Summary Incubation of conidia of wild typeNeurospora crassa at temperatures ranging from 25 to 46°C modulates their respiratory type. Between 37 and 41°C, the transient activity of the cyanide-insensitive respiratory pathway parallels, with a maximal extrusion of protons into the medium, the optimal rate of germ tube outgrowth.We are grateful to Mrs Arlette Cattaneo for her competent technical assistance.     

Kälte als Auslöser einer Glykogen-Speicherung während der Oogenese vonMusca domestica

Summary InMusca domestica reared under standard conditions (21 °C), glycogen is stored in the oocyte towards the end of egg development. By 1–3 days exposure to lower temperatures (4 °C), a glycogen deposition can be released already in young follicles. This premature glycogen synthesis is not restricted to the ooplasm. Carbohydrates are also found in the nurse cells and the follicle epithelium. Similar results were formerly obtained by inhibition of oogenetic protein synthesis.

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Herrn Prof. Dr. Dr. h.c.Bernhard Rensch zum 70. Geburtstag gewidmet.     

Effect of relative humidity on the free-living stages ofStrongyloides papillosus (Rhabdiasoidea: Nematoda)

Summary The eggs and larvae are affected by the changes in the humidity level around them. The eggs do not develop below 87% relative humidity at 30 °C and 25 °C. At sub-developmental rel. hum. they remain viable for 30 h at 81%, but at 73% rel. hum. level they do not survive beyond 18 h. Survival of larvae in 100% rel. hum. is longer at 30 °C than at 25 °C and 35 °C, and also they exhibit a poor resistance to desiccation.Acknowledgment. The author is grateful to Dr. Devendra Prasad of Patna University for his supervision and constant help in the work. The assistance of State University Grants Commission through a research scholarship is gratefully acknowledged.     

The effect of chronic heat stress on cortisol levels in the antarctic fishPagothenia borchgrevinki

Radioimmunoassay was used to determine levels of the stress-inducible glucocorticoid, cortisol, circulating in the plasma of the extremely stenothermal Antarctic fishPagothenia borchgrevinki at rest and after heat stress. Fish sampled immediately after capture (–1.9°C) had low cortisol levels (10.4±1.4 ng ml^–1, mean±SEM) as did fish which were laboratory rested for 3 days. Sudden exposure to 5°C (48h) resulted in a peak cortisol value after 3 h (69.9±6.8 ng ml^–1) whereas exposure to 8°C (6h) resulted in a peak value after 1 h (73.5±8.0 ng ml^–1). At both temperatures levels remained significantly elevated (p<0.05) for the entire period of exposure. Increased temperature also resulted in a significant change in haemoglobin, haematocrit and mean cell haemoglobin concentration (MCHC) (p<0.05). Plasma lactate was significantly elevated only after exposure to 8°C (p<0.05). Plasma cortisol levels fromP. borchgrevinki are reported here for the first time and show this cryopelagic Antarctic species to have an unusual hormonal stress profile.     

Growth ofEchinococcus multilocularis in gerbils exposed to different environmnntal temperature

Summary Gerbils kept at high (35°C) temperature harbored a significantly largerEchinococcus multilocularis cyst biomass than those kept at room (21°C) and low (5°C) temperature. Parasite induced adrenomegaly was observed in all temperature groups.Acknowledgments. I am indebted to Miss Marianne Hardy and Mr Terry Durham for their excellent technical assistance.     

Effects of low temperatures on survival of frozen-thawed mouse embryos

Summary Almost no damage to mouse morulae was observed between 0 and –40°C, and freezing damage to embryos in DMSO, glycerol or ethylene glycol occurred after exposure to –60, –50 or –50°C, respectively. Cooling embryos in DMSO or glycerol to temperatures below –50 to –60°C increased freezing damage. To the contrary, in the presence of ethylene, glycol, no more damage occurred after exposure to temperatures below –50°C.     

Ventilatory responses to CO2 at different body temperatures in the snake,Coluber constrictor

Summary Ventilatory responses to CO₂ were examined at different temperatures in the snake,Coluber constrictor. CO₂ sensitivity increased between 15 and 25°C but not between 25 and 35°C. A rapidly occurring off-CO₂ transient hyperpnea suggested the presence of an intrapulmonary chemoreceptor.     

Melezitase and maltase from the midgut ofSesamia inferens Walker (Lepidoptera: Insecta)

Summary The activity of melezitase and maltase was optimal at pH 6.2 and temperature 35–40°C and inhibited by the end-products. Melezitase activity was affected by K⁺, Li⁺⁺ andTris ions and was reduced by dialysis.Acknowledgments: The author is thankful to ProfessorR. Rakshpal for valuable guidance, and to University Grants Commission, for awarding me a Junior Research Fellowship.     

Antifreeze agents in the body fluid of winter active insects and spiders

Summary Insects and spiders which are active at subzero temperatures on snow in winter are found to be protected against internal freezing by antifreeze agents present in their body fluid. The body fluid has a melting point of about –1 °C, but the antifreeze agents prevent growth of ice crystals at temperatures down to –6 to –7 °C.Acknowledgment. We would like to thank Anne Lohrman for her kind help in collecting insects and spiders.     

The effects on thermoregulation of intracerebroventricular injections of L-aspartic acid in the sheep

Summary L-Aspartic acid was injected into a lateral cerebral ventricle of the sheep at ambient temperatures between 0° and 40°C. Doses of 100 or 500 nmoles·kg^–1 caused a rise in heat production and/or a decrease in heat loss; rectal temperature rose. Atropine sulphate attenuated or prevented these effects.     

Motility of leukocytes in slide cells

Zusammenfassung Die Leukozytenbewegung wurde in Glaskammern während 24 h Inkubation beobachtet: die Kurve der Leukozytenbewegung besteht aus 2 Phasen, welche mittels 2 Exponentialgleichungen (durch das Wachstumsgesetz bestimmt) ausgedrückt werden können.In der vierten Stunde der Inkubation tritt eine Steigerung der Leukozytenbewegung auf.Die Leukozytenbewegung bei einer Temperatur von 30 und 40°C wurde mit der bei 37°C verglichen: Die Leukozytenbewegung ist bei 30°C langsamer und bei 40°C schneller als bei 37°C.     

Hibernation at moderate temperatures: a continuation of slow wave sleep

Summary Golden-mantled ground squirrels (Citellus lateralis) displayed virtually continuous electrophysiological states of sleep when hibernating at moderate ambient temperatures (22°C). Rapid-eye-movement sleep progressively diminished with the fall in body temperature so that at a body temperature of 23°C it was completely absent. At this temperature hibernation was characterized by slow wave sleep isomorphic with slow wave sleep episodes at non-hibernating (euthermic) body temperatures.Supported by National Institute of Health grants GM 23694 awarded to R.J. Berger and GM 23695 awarded to H.C. Heller.     

Veränderungen der thermoelastischen Kontraktion von Haut und Nerv bei alternden Tieren

Summary The thermo-elastic contraction which has been found to increase with age in the tendons, has been studied in the skin. This shows a contraction between 64–66°C similar to that of tendon fibres. In 22 month old rats, it reaches 5–6 times greater values than in 3 month old animals. The skin also shows a second contraction between 67 and 70°C. After the ninth month, in a continuously increasing degree, a third and fourth step of contraction appear at higher temperatures (up to 82°C).The sciatic nerve also shows a thermo-elastic contraction after the fourth month at 76–80°C. This increases with age, and, as in the case with the skin, several more steps of contraction appear at higher temperatures, up to 92°C. It is improbable that these thermo-elastic contractions are related to «elastin». The changes with increasing age, however, show a similarity to the age changes of «elastin» in so far as the thermic contractility increases with age.     

Elucidation of the relationship between enzyme activity and internal motion using a lysozyme stabilized by cavity-filling mutations

We investigated the activity and the internal motions of a stabilized mutant hen lysozyme (HEL) in which the residues M12 and L56 were mutated to L and F, respectively (LF mutant HEL). The result of the activity measurements against glycol chitin at various temperatures suggested that the temperature dependence of the activity of LF mutant HEL shifted to the high-temperature side compared with that of wild-type HEL. The detailed internal motions of LF mutant HEL in the absence and presence of a substrate analogue, (NAG)₃, were examined by model-free analysis at 35°C. The results showed that the internal motions of LF mutant HEL in the presence of (NAG)₃ were drastically restricted compared with those in wild-type HEL. Our findings thus suggested that the mutation to the stabilized lysozyme restricted internal motions required for the enzymatic reaction.Received 8 February 2005; accepted 10 March 2005Y. Yoshida and T. Ohkuri contributed equally to this work.