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Dynamic of the genetic structure of bacterial and fungal communities at different developmental stages of Medicago truncatula Gaertn. cv. Jemalong line J5.

The genetic structure of bacterial and fungal communities was characterized in the rhizosphere of Medicago truncatula Gaertn. cv. Jemalong line J5 at five developmental stages (three vegetative and two reproductive stages), and in three compartments (bulk soil, rhizosphere soil and root tissues). The genetic structure of microbial communities was determined by cultivation-independent methods using directly extracted DNA that was characterized by automated ribosomal intergenic spacer analysis (ARISA). Principal component analyses (PCA) indicate that, for all developmental stages, the genetic structure of microbial communities differed significantly by compartment, with a major shift in the community in root tissues corresponding to the most intimate compartment with the plant. Differences were also recorded during plant development, the most significant being observed during the transition between vegetative and reproductive stages. Throughout this period, plants were shown to establish the highest level of symbiotic association (mycorrhization, nodulation) with arbuscular mycorrhizal fungi and Rhizobia. During the reproductive stages, the dynamics of the genetic structure differed between bacterial and fungal communities. At the last reproductive stage, the genetic structure of bacterial communities became close to that recorded during the first vegetative stages, suggesting a resilience phenomenon, whereas the genetic structure of fungal communities remained different from the vegetative stages and also from the early reproductive stages, suggesting a persistence of the rhizosphere effect.

Bacteria↗

Microtubule-associated proteins present in different developmental stages of Drosophila melanogaster.

Microtubule-associated proteins (MAPs) have been isolated from different development stages of Drosophila melanogaster and characterized by their association to tubulin, but not to tubulin lacking its 4-kD carboxy terminal region (S-tubulin), and by their ability to promote tubulin polymerization. Following these criteria some peptides of Mr 255, 205, and 180 kD were identified as MAPs. By means of immunological analogy we have identified a peptide related to mammalian brain MAP known as tau factor.

Animals↗

Molecular analysis of microbial communities identified in different developmental stages of Ixodes scapularis ticks from Westchester and Dutchess Counties, New York.

Ixodes scapularis ticks play an important role in the transmission of a wide variety of pathogens between various mammalian species, including humans. Pathogens transmitted by ticks include Borrelia, Anaplasma and Babesia. Although ticks may harbour both pathogenic and non-pathogenic microflora, little is known about how the diversity of the microflora within ticks may influence the transmission of pathogens. To begin addressing this question, we examined the composition of bacterial communities present in Ixodes scapularis collected from Westchester and Dutchess Counties, New York State, at different developmental and nutritional stages. Genetic fingerprints of bacterial populations were generated by temporal temperature gradient gel electrophoresis (TTGE) separation of individual polymerase chain reaction (PCR)-amplified 16S rRNA gene fragments, followed by DNA sequence analysis for bacterial identification. The fingerprints of the TTGE bands were grouped into five clusters. The most abundant DNA sequence found in all the samples was Rickettsia, followed by Pseudomonas and Borrelia. Ralstonia, Anaplasma, Enterobacterias, Moraxella, Rhodococcus and uncultured proteobacterium were present as well. We also determined the prevalence of Anaplasma phagocytophilum and Borrelia burgdorferi by PCR and DNA sequence analysis. Statistical analyses indicated significant variations in the bacterial communities depending on tick developmental stage and degree of engorgement. We suggest that these two elements affect microbial diversity within the tick and may in turn influence pathogen transmission to humans and animals after tick bite.

Animals↗

Ethoxyresorufin-O-deethylase (EROD) inducing potencies of planar chlorinated aromatic hydrocarbons in primary cultures of hepatocytes from different developmental stages of the chicken.

In vitro induction of ethoxyresorufin O-deethylase (EROD) activity in cell cultures is an extensively validated tool for measuring overall potencies of mixtures of halogenated aromatic hydrocarbons (HAHs) in samples from the abiotic or biotic environment. For risk assessment with special attention to effects in wild birds, an assay was developed that makes use of chicken embryo hepatocytes. However, it was questioned whether compound-specific responses are consistent at the various developmental stages. The results of our present study show that there are considerable differences between early and late embryonal and post-hatching stages. The induction of EROD was measured in primary chicken hepatocyte cultures. The cells were isolated at day 14 and day 19 of embryonal development and at day 1 post hatching. Hepatocytes were exposed in vitro to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 3,3',4,4',5-pentachlorobiphenyl (PCB 126, IUPAC nomenclature) and 2,3',4,4',5-pentachlorobiphenyl (PCB 118). The respective compounds were chosen as representatives for dioxins, furans, non-ortho PCBs, and mono-ortho PCBs. These groups of chemicals have been identified as environmental contaminants with major dioxin-like effects that are mediated by a common receptor, the arylhydrocarbon (Ah) receptor. At all developmental stages, TCDF was more potent than TCDD. Relative potencies (RP = EC50TCDD/EC50HAH) decreased in the order TCDF < TCDD < PCB 126 < PCB 118. Depending on the developmental stage, TCDF was 1.2 to 3.4 times more potent than TCDD. PCB 126 was equipotent or less potent by a factor of 3 than TCDD. PCB 118 was 100 to 300 times less potent than TCDD. Both the mean effective concentration (EC50) and the maximum EROD activity (Ymax) of all compounds were lower in hepatocyte cultures from 14-day-old embryos than those from 19-day-old embryos or 1-day-old hatchlings. RPs were comparable in 19-day-old embryos and in hatchlings, but significantly different in 14-day-old embryos.

Animals↗

Brain and pituitary beta-endorphin levels at different developmental stages of the rat.

beta-Endorphin-like immunoreactivity in whole brains of Sprague-Dawley rat fetuses of different gestational ages was measured by radioimmunoassay and found to increase throughout the gestational period studied. The immunoreactivity in various brain parts (forebrain, midbrain, hindbrain, hypothalamus and pituitary) of late prenatal, early postnatal, young mature and retired breeder rats was also determined. In all the brain parts studied, a maximum in the content and concentration of beta-endorphin-like immunoreactivity was attained when the rats were about 3-4 months old.

Aging↗

Double vitrification of rat embryos at different developmental stages using an identical protocol.

The aim of the present investigation was to test the effectiveness of a method of vitrifying rat embryos at different stages of development (from early morula to expanding blastocyst) in a double vitrification procedure. Wistar rat embryos were vitrified and warmed in super-fine open-pulled straws (SOPS). Before being plunged into liquid nitrogen, the embryos were exposed to 40% ethylene glycol+0.75 M sucrose in TCM-199+20% fetal calf serum (FCS) for 20s at 38 degrees C. Subsequent warming and direct rehydration of the embryos was conducted in culture medium (TCM-199+20% FCS) at 38 degrees C. Early morula stage (7-10 blastomeres) embryos (n=358) were vitrified, warmed and cultured in vitro (EM group). Batches of these embryos were then cryopreserved again (revitrified) at the early blastocyst (EB group, n=87), blastocyst (B group, n=93) or expanding blastocyst stage (ExpB group, n=73). After the first (EM group) and repeated (EB, B, and ExpB groups) vitrification procedures, developmental rates of 81, 83, 34 and 76%, respectively were achieved (for EM-EB-ExpB P>0.1; for EM, EB, ExpB-B P<0.005). Our data demonstrate the possibility of using the described identical protocol for the SOPS vitrification of rat early morulae, early blastocysts and expanding blastocysts. The low survival rate of blastocysts subjected to double vitrification requires further investigation.

Animals↗

Toxicity of the venom from Nasonia vitripennis (Hymenoptera: Pteromalidae) toward fly hosts, nontarget insects, different developmental stages, and cultured insect cells.

A venom preparation from Nasonia vitripennis, a wasp ectoparasitoid of fly pupae, was assayed for lethality in different stages of insects representing ten different orders and in cultured insect cells. In most cases, the motor activity of the injected insects remained completely normal for 1-2 days after the injection and displayed none of the symptoms of paralysis commonly reported for venoms of the Hymenoptera. A natural host, the flesh fly Sarcophaga bullata, was highly sensitive in the pupal stage (LD50 = 5.4 and 5.5 VRE/g for nondiapausing and diapausing pupae, respectively), the stage that is normally parasitized, and larvae and adults were as susceptible to the venom as the pupae. Adults of another fly host, Phaenicia sericata, were nearly as sensitive (LD50 = 6.5 VRE/g), but nonhost adult flies were more tolerant. Among the other orders tested, pupae of several species (Plodia interpunctella, Trichoplusia ni, Tenebrio molitor) were more susceptible to envenomation than larval or adult stages. In fact, the highest sensitivity observed in this study (LD50 = 0.58 VRE/g) was with pupae of the cabbage looper, T. ni, a species that is not a natural host. In contrast, the larvae (LD50 = 7.23 VRE/g) and adults (LD50 = 7.48) of T. ni were far less sensitive. Adults of Nasonia vitripennis were not sensitive to their own venom (LD50 = > 533 VRE/g), although adults of another hymenopteran, Apis mellifera, were suceptible (4.62 VRE/g). Adults of Lymantria dispar, Oncopeltus fasciatus, Aphis nerii, Euborellia annulipes, Diapheromera femorata, Blattella germanica, Periplaneta americana, and Reticulitermes flavipes demonstrated a high tolerance to Nasonia venom. When tested in vitro, the venom caused cultured Lepidoptera (TN-368) and Diptera (NIH SaPe4) cells to round up, swell, and eventually die. The LC50S were 0.0014 and 0.0010 VRE/microliters for TN-368 and SaPe4 cells, respectively. Cytotoxicity was observed within 10 min after exposure to LC99 levels of venom, with 100% cell mortality at 100 min for the NIH SaPe4 cells and 24 hr for TN-368 cells. It is possible that the venom component responsible for in vivo and in vitro activities may be different, but results from the cell culture work suggest that this method offers a promising assay for quickly screening venom samples. The high susceptibility of flies and pupae of other insects to the venom, as well as its novel (nonparalytic) action suggest that it may have considerable potential for development as a biopesticide.

Animals↗

[The correlation between the heat resistance of the zygotes of the common frog Rana temporaria and its embryos at different developmental stages].

A study has been made of a correlation between the heat resistance of zygotes and embryos of the same clutches at different stages of development of Rana temporaria L. The embryos were incubated at 19 degrees C, the injurious temperature being 37 degrees C. As criterion of heat resistance served the time of the injurious temperature action which leads by the ++stra of cleavage to a 50 per cent elimination of embryos (LD50). The correlation in question has been evaluated by the rank correlation coefficient (rho). The correlation between the heat resistance of zygotes and embryos at the start of gastrulation (stage 11) was moderate (rho = 0.47). At one of neurulation stages (stage 22) this correlation was weak (rho = 0.207), to become rather high just before the cleavage (stage 28, rho = 0.63). It is assumed that the thermal selection of zygotes, i.e. of the organism at the cellular stage of development, may be oriented to result in the survival of embryos, which just before the cleavage will show both elevated heat resistance and high heritability of this character.

Animals↗

The messenger RNA for alcohol dehydrogenase in Drosophila melanogaster differs in its 5' end in different developmental stages.

Alcohol dehydrogenase (EC 1.1.1.1) of Drosophila melanogaster is coded by a single structural gene, active in both larvae and adults. The major larval and adult transcripts of Adh differ in their 5'-untranslated regions. The major larval mRNA is about 1100 bases long, some 50 bases shorter than the major adult transcript. The 5' end of the larval mRNA is colinear with the genomic sequence immediately adjacent to the coding region, starting 70 base pairs (bp) upstream of the initiation codon. By contrast, the adult mRNA shares only 36 of its 123 5'-untranslated bases with the larval mRNA; the remaining 87 are encoded by a sequence 654 bp upstream. Both initiation sites are preceded by a TATA box some 24 bp upstream. The developmental specificity of Adh expression is seen, therefore, to have a counterpart in the specificity of transcription initiation at the two separate promoter regions.

Alcohol Dehydrogenase↗

[Interaction of cis-regulatory element in the 5' flanking sequence of human beta-globin gene with trans-acting factors at different developmental stages].

Human beta-globin gene mainly expressed in the adult bone marrow, while not expressed in fetal liver, adult liver and K562 cells. Using gel mobility shift assay, different protein factors binding to the regulatory elements (from -372 to -194 bp) in the 5' flanking sequence of human beta-globin gene were detected in the nuclear extracts from human fetal liver, adult liver and K562 cells respectively. Competitive experiments showed that both the protein factors from adult and K562 cells not only bind to negative control region 2 (NCR 2, from -372 to -224 bp), but also bind to positive control region (PCR, from -223 to -194 bp), suggesting that there may be similar mechanism to silence the expression of human beta-globin gene in these two kind of cells. The protein factor in human fetal liver only binds to NCR 2, indicating that there is a particular silencing mechanism for beta-globin gene expression in human fetal stage.

Fetus↗

[DNA apurinization and apyrimidinization of gamma-irradiated silkworm embryos at different developmental stages].

A study was made of the formation and repair of apurine-apyrimidine (AP) sites in DNA of gamma-irradiated 3- and 7-day embryos of Bombyx mori differing drastically in radiosensitivity. The number of AP-sites in DNA immediately after irradiation was much larger in DNA of 3-day embryos than in DNA of 7-day embryos. The kinetics of the postirradiation recovery of AP sites in DNA of 3- and 7-day Bombyx mori embryos was heterogeneous and varied significantly.

Animals↗

Two-dimensional protein maps of xenopus eggs and embryos at different developmental stages.

Protein expression during the early development of Xenopus has been followed by 2D-polyacrylamide gel electrophoresis (PAGE). The analysis of two-dimensional maps of eggs and embryos at different stages of development has allowed the separation of more than 2000 spots. Identification of numerous polypeptides was obtained in four different ways: (1) immuno-blotting; (2) amino terminal sequence after blotting on to PVDF membranes; (3) comigration; and (4) assignment in comparison with proteins separated by 2D techniques on reference maps such as human liver, red blood cells, plasma and cerebrospinal fluid reported in the Swiss 2D-PAGE Data Base. The maps presented in this report are a step toward the study of the protein expression in Xenopus eggs and embryos and may be a powerful working tool since Xenopus embryos are popular models for the study of development.

Animals↗

Effect of heat on radiosensitivity at different developmental stages of embryos of the fish Oryzias latipes.

Embryos of the fish, Oryzias latipes, at various developmental stages were heated before or after gamma-irradiation. The lethal effects of heat and radiation treatments were determined by the hatchability of the irradiated embryos. The radiosensitivity suddenly decreased at 3-days after fertilization. Heat (41 degrees C), either preceding or following irradiation, decreased the shoulder region (Dq) of the dose-response curves of the embryos irradiated at the 1-3 day stages, and increased the slope (D0) only at the 3 day stage. The magnitude of enhancement of radiosensitivity by pre- and post-heating was found to depend on the time and temperature of the heat as well as the developmental stage of the embryos; in 1- and 2-day embryos the effectiveness of post-irradiation heating was larger than that of pre-irradiation heating whereas in 3-day embryos there was no difference between the effectiveness of pre- and post-treatment. The sequence of heat (41 degrees C) and radiation treatment, and the time interval between the two treatments, also influenced the effectiveness of heat on radiosensitivity, depending upon the developmental stage; recovery from heat effects was not observed within a time interval of up to 3 h before radiation exposure for either 1- or 3-day embryos, whereas recovery from the radiation lesion occurred, for only 1-day embryos, within 1 h of subsequent heat exposure.

Age Factors↗

Degenerative neuronal changes in the rat thalamus induced by status epilepticus at different developmental stages.

SE was induced in Wistar rats at post-natal (P) days 12, 15, 18, 21, and 25 to determine distribution and severity of thalamic damage in relation to time after SE. Six different intervals from 4 h up to 1 week were studied using Fluoro-Jade B (FJB) staining. Severity of damage was semi-quantified for every age-and-interval group. Distribution of neuronal damage within various thalamic nuclei was mapped by a computer-aided digitizing system. A consistent neuronal damage occurred in functionally heterogenous thalamic nuclei. Damage was found in all age groups although its extension and time course as well as the number of involved thalamic nuclei varied. Number of injured thalamic nuclei rapidly increased with age on SE-onset. In P12 group, degenerating neurons were consistently seen in the mediodorsal and lateral dorsal thalamic nuclei. Since P15, neurodegeneration was observed additionally in midline, ventral and caudal thalamic nuclei (visual and auditory thalamic nuclei), in the lateral posterior and in the reticular nucleus. In P21 and P25 animals, the majority of thalamic nuclei exhibited marked neuronal damage. Nuclei with a small number (anterior and intralaminar) or no FJB-positive neurons (the ventral nucleus of the lateral geniculate body) were exceptional. The pattern of thalamic damage is age-specific; its extent and severity increases with age.

Age Factors↗

The effect of salinity on different developmental stages of an endemic annual plant, Aster laurentianus (Asteraceae).

Salinity reduces substrate water potential, thereby restricting water and nutrient uptake by plants; salinity may also cause ionic imbalance and toxicity. Because substrate salinity fluctuates through the growing season, a plant may be exposed to different salinity levels, at various stages of development, with potentially significant consequences on population dynamics. Here, we present the results of a study of the effect of substrate salinity on seed germination, seedling emergence, and growth of Aster laurentianus, an annual marsh plant, endemic to the Gulf of St. Lawrence and potentially threatened. Seed germination was reduced in low salt concentration (10 g sea salt/L) and completely inhibited by salinity levels >/=20 g sea salt/L. However, this inhibiting effect was reversible: seeds from the salt treatments germinated readily after being washed in distilled water. Though seedling emergence was diminished at low salinity levels, postemergence survival was little affected. Plant growth was reduced, but net carbon assimilation rate was not affected by high salinity levels. Increased root respiration and respiratory costs associated with salt tolerance might have contributed to lower C accumulation at higher salinity levels. All developmental processes considered are thus negatively affected by substrate salinity, with potentially significant consequences on population abundance and distribution in salt marshes. Yet, the tolerance of this species to high salinity levels after seedling emergence is remarkable. Seed germination represents a major bottleneck in the species life cycle, potentially controlling local distribution and abundance in the natural habitat.

Journal Article↗

Hemicerebellectomy and motor behaviour in rats. II. Effects of cerebellar lesion performed at different developmental stages.

Rats with a right hemicerebellectomy (HCb) performed in adulthood or at weaning were compared behaviourally to rats with a similar lesion performed on the first postnatal day. The age at which animals received cerebellar lesions made a significant difference with respect to the behavioural outcome in adulthood. Posture, locomotion and motor behaviour were analysed by a battery of sensorimotor tests. Behavioural measurements showed a clear relationship between age at surgery and behavioural effects; rats with neonatal cerebellar lesions showed a slight extensor hypotonia contralateral to the lesion side and efficient locomotor activity, while the adult operated group exhibited a severe extensor hypotonia ipsilateral to the lesion side and hampered locomotion characterized by a wide base and ataxia. Weanling operated rats displayed a symptomatology similar to that observed in adult operates, although less severe. In the postural dynamic adjustments which the sensorimotor tests required, the youngest operated animals obtained higher scores in comparison to the other two experimental groups, except for the lack of hindlimb usage in the suspension on a wire test. These results, which show the importance of the age-at-lesion factor for the recovery of motor function after HCb in the rat, are discussed in the light of the widespread anatomical reorganization already demonstrated following neonatal HCb in rats.

Aging↗

[A comparative study of the DNA structural damages and synthesis in embryos of the silkworm Bombyx mori irradiated with gamma rays at different developmental stages].

Studies have been made on formation and reparation of apurine-apyrimidine (AP) regions, monothread DNA ruptures, as well as on inhibition and recovery of DNA synthesis in gamma-irradiated 3- and 7-day embryos of the silkworm which sharply differ in their radiosensitivity. It was shown that in 3-day embryos, the number of AP regions and DNA ruptures immediately after irradiation is significantly higher than in 7-day embryos. DNA synthesis is more radiosensitive in 3-day embryos as compared to that in 7-day ones. Kinetics of postradiation recovery of regions and DNA ruptures in 3- and 7-day embryos is heterogeneous and significantly different. However, radiation inhibition and postradiation recovery of DNA synthesis in irradiated 3- and 7-day embryos are associated mainly with postradiation survival of these embryos.

Animals↗

Study and characterization of different developmental stages of Leishmania major.

The complete developmental cycle of Leishmania major in axenic culture was achieved by simply changing the temperature whether sudden from 22 degrees C to 37 degrees C or stepwise 22 degrees C, 29 degrees C and then 37 degrees C. The morphology by light microscopy, GPI isoenzyme pattern and PCR amplification of minicircle of kinetoplast DNA of the different stages were studied. The amastigotes obtained from the foot pads of mice were compared to those obtained from axenic culture. The GPI isoenzyme pattern and the PCR amplification products showed distinct differences between the promastigotes and the amastigotes. The amastigotes of the two sources also showed differences after temperature changes.

Animals↗