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Results for “Developmental stage”

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PubMed1975

The ribosomal proteins of drosophila melanogaster. IV. Characterization by two-dimensional gel electrophoresis of the ribosomal proteins from nine postembryonic developmental stages.

The ribosomal proteins from nine postembryonic developmental stages of Drosophila melanogaster were analyzed by two-dimensional gel electrophoresis. The ribosomal protein patterns thus obtained constitute reliable control patterns against which any potential mutant can be compared. Furthermore, the results provide increased evidence for a qualitative and quantitative change in the protein composition of the total population of ribosomes which occurs mainly during the third larval instar. Two acidic proteins were found, which may be homologous with rat liver ribosomal proteins L40/L41 and E. coli proteins L7/L12.

Animals↗

PubMed1990

Developmental-stage-specific plasmid supercoiling in Chlamydia trachomatis.

Chlamydia trachomatis elementary body (EB) and reticulate body (RB) developmental stages have polymorphic plasmid DNA. Several plasmid forms separated by gel electrophoresis were identified as topoisomers by treatment with topoisomerase I. Among these topoisomers was one form unique to EBs and one form unique to RBs. The unique EB plasmid topoisomer was characterized as highly supercoiled, on the basis of band migrations by gel electrophoresis and its appearance by electron microscopy. The unusual physical state of this topoisomer was probably mediated, in part, by DNA-specific structural proteins. The unique RB plasmid topoisomer was a supercoiled form of lower superhelical density than the other identified topoisomers. Developmental-stage-specific differences in super-helical density of plasmid DNA suggest cause-and-effect relationships between DNA topology and metabolic activity in RBs and metabolic quiescence in EBs.

Blotting, Southern↗

PubMed2006

Profiling of maternal and developmental-stage specific mRNA transcripts in Atlantic halibut Hippoglossus hippoglossus.

cDNA libraries were constructed from the following developmental stages (tissues) of the Atlantic halibut (Hippoglossus hippoglossus): 2-cell stage (embryos), 1 day-old yolk sac larvae (trunk) and juvenile (fast skeletal muscle). A total of 4249 high quality expressed sequence tags from the three libraries were clustered into a partial transcriptome of 2124 putative genes. A large proportion of the gene clusters (48.3%) had no significant matches against known proteins. The most abundant ESTs of nuclear transcripts in the 2-cell library included sequences with high identity to zebrafish H1M, a linker histone-like protein involved in primordial germ cell specification, zinc finger protein, rRNA external transcribed spacer, thymosin beta-4, cyclin B1 and several predicted peptides from the Tetraodon nigroviridis genome assembly with unknown functions. 170 and 123 ESTs represented ribosomal proteins in the larval and juvenile libraries respectively, compared with only two sequences in the 2-cell library, which may reflect an abundance of maternally inherited pre-formed ribosomes in the yolk. Even though some clusters were common to all three libraries, most putative genes showed a developmental-stage specific distribution with 72% (2-cell embryo), 59% (larval) and 57% (juvenile) sequences having no significant matches against the 8400 adult halibut sequences in the EMBL nucleotide database. Comparison between the predicted halibut peptide data set and the human, zebrafish, and pufferfishes (T. nigroviridis and Takifugu rubripes) proteomes revealed that, as expected, the halibut sequences were more similar to the other two fish species than to human proteins. However, no clear bias towards the pufferfishes was observed, suggesting significant sequence variation between orthologues within the clade Acanthomorpha. The sequence information generated in the present study will represent a significant new resource for future studies on normal and abnormal development in Atlantic halibut.

Animals↗

PubMed1992

Developmental stages and energy restriction affect cellular oncogene expression in tissues of female rats.

The influence of developmental stages and energy restriction on c-Ha-ras and c-fos cellular oncogene mRNA level was examined in mammary tissue and liver of female rats. c-Ha-ras and c-fos RNA transcripts in mammary tissue peaked at 9 wk of age, decreased during pregnancy, were lowest at mid-lactation and then increased at 25 wk. c-Ha-ras in liver showed highest expression at 5 wk and was hardly detectable at 11 wk, mid-pregnancy, mid-lactation and 25 wk. Liver c-fos and c-Ha-ras mRNA levels were similar. Lower c-Ha-ras and c-fos mRNA levels were observed in mammary tissue of energy-restricted (to 70% of ad libitum intake) rats at 25 wk compared with those allowed ad libitum access to food. Energy restriction did not affect liver c-Ha-ras and c-fos mRNA levels throughout the experimental period. These results demonstrate that developmental stages and energy restriction affect cellular oncogene mRNA levels with tissue-specific patterns.

Aging↗

PubMed1997

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↗

PubMedDate not supplied

Comparative EST analyses provide insights into gene expression in two asexual developmental stages of Eimeria tenella.

The protozoan parasite Eimeria tenella has a complex life cycle that includes two major asexual developmental stages, the merozoite and the sporozoite. The expressed sequence tag (EST) approach has been previously used to study gene expression of merozoites. We report here the generation and analysis of 556 ESTs from sporozoites. Comparative analyses of the two datasets reveal a number of transcripts that are preferentially expressed in a specific stage, including previously uncharacterised sequences. The data presented indicate the invaluable potential of the comparative EST analysis for providing information on gene expression patterns in the different developmental stages of E. tenella.

Animals↗

PubMed1992

[Antigenic localities in the tissues of Paragonimus westermani by developmental stages using immunogold labeling method].

In order to observe the antigenic localization in the tissues of Paragonimus westermani of developmental stages, immunogold labeling method was applied using serum of the cats which were infected with isolated metacercariae from Cambaroides similis. The sectioned worm tissues from each developmental stage were embedded in Lowicryl HM 20 medium, stained with infected serum IgG and protein A gold complex (particle size: 12 nm) and observed by electron microscopy. In the young adult worm tissue of 4 weeks after infection with metacercariae, the gold particles were specifically concentrated on the tegumental syncytium and cytoplasm of the tegumental cells as well as the secretory granules in the parenchymal tissue. The antigenic materials in the adult worm tissue were specifically concentrated on the secretory granules in the parenchymal tissue, the cytoplasm between granules in the vitelline gland and the epithelial lamella in the lumen of the caecum.

Animals↗

PubMed2005

Characterisation of carbohydrate-binding sites in developmental stages of Myxobolus cerebralis.

Glycans and lectins (carbohydrate-binding molecules) form a mutual recognition system, which enables parasitic organisms to attach themselves to the host cells and/or take part in the migration of their developmental stages into the target tissue. The aim of the present study was to identify and characterise the potential binding activity of glycoconjugates in different developmental stages of Myxobolus cerebralis, the causative agent of whirling disease in salmonids. The binding patterns of 13 biotinylated neoglycoconjugates were histochemically examined in thin-sections of infected rainbow trout (Oncorhynchus mykiss) and oligochaetes (Tubifex tubifex), as well as isolated waterborne triactinomyxon spores. A distinct structure-selective and developmental stage-regulated expression of certain classes of carbohydrate binding was observed. In triactinomyxon spores, the expression of carbohydrate binding activity for alpha-l-Fuc-BSA-biotin, alpha-d-GalNAc-BSA-biotin, beta-d-GlcNAc-BSA-biotin, Lac-BSA-biotin and ASF-biotin was up-regulated in the polar capsules; the shell valves showed no activity. In the gut of T. tubifex, polar capsules of the parasite showed strong positive reaction only for beta-d-GlcNAc-BSA-biotin. In fish cartilage, polar capsules were negative, but the spore shell valves showed a broad range of carbohydrate binding activity. No activity was detected for either alpha6- or alpha3-linked N-acetyl-d-neuraminic acid to galactose. An adhesion assay was performed on GlycoWell plates and Myxobolus spores were found to specifically adhere to matrices containing residues of lactose, fucose, galactose, N-acetyl-d-galactosamine and N-acetyl-d-glucosamine. This is the first study to identify lectin activity in a myxozoan parasite; activity that is likely to play a role in the recognition systems involved in host specificity and the processes of spore attachment and invasion.

Animals↗

PubMedDate not supplied

Trypanosoma cruzi: flow cytometric analysis of developmental stage differences in DNA.

Flow cytometry and DNA binding-specific fluorescent reagents were used to compare the total DNA, G-C, and A-T content of the epimastigote and trypomastigote stages of Trypanosoma cruzi stocks. Significant total DNA differences of 2-12% between epimastigotes and trypomastigotes were found in three of six stocks studied. The epimastigote G-C content of five of six stocks was 4-8% higher than trypomastigotes, whereas the trypomastigote A-T content was 2.5-13% higher than the epimastigote A-T content. Although no obvious developmental stage association between total DNA and base composition was found, intrastage associations do exist. These observations were unaffected by nucleoprotein extraction implying that the observed differences between trypomastigotes and epimastigotes are not a consequence of nucleoprotein interference with DNA-binding fluorochromes. The nuclei and kinetoplasts of four T. cruzi stocks were isolated and analyzed. Developmental stage differences in nuclear and kinetoplast DNA are stock-dependent and base composition-dependent; both organelles contribute to the observed differences in DNA of intact cells. We found a nearly linear association between the percentage of total kinetoplast DNA, G-C, and A-T content. During metacyclogenesis, the G-C content decreases by approximately 7% as epimastigotes transform into metacyclic trypomastigotes. The decrease in G-C content precedes changes in morphology or in complement resistance. If the DNA changes are causally connected to developmental stage transformations in T. cruzi remains to be determined. However, our results could facilitate studies of the molecular genetic processes the parasite uses to successfully complete various phases of its life cycle and, consequently, the disease process it evokes.

Analysis of Variance↗

PubMed1994

Effects of quality and developmental stage on the survival of IVF-derived bovine blastocysts cultured in vitro after freezing and thawing.

Factors affecting viability of IVF-derived bovine blastocysts after freezing and thawing were investigated. A total of 1,101 ova matured and fertilized in vitro were cultured under 2 different conditions, 1) in TCM-199 on granulosa cell monolayers at 5% CO(2) in air and 2) in synthetic oviduct fluid (SOF) medium without somatic cell support at 5% CO(2), 5% O(2), 90% N(2). All blastocysts that developed from the 2 different culture systems were individually classified into 4 grades of embryo quality and were then frozen by conventional slow freezing. Developmental rates of the IVF-derived ova to blastocysts and the survival rates of the frozen-thawed blastocysts were not different between the SOF medium (16 and 49%) and the co-culture system (13 and 61%, respectively). Survival of frozen-thawed blastocysts was affected by embryo quality in both the SOF and co-culture systems (P<0.001). Blastocysts produced in vitro were also individually classified into 3 developmental stages and were then cultured for 3 d in the co-culture system with granulosa cells after freezing and thawing. There was a difference in the survival rate of frozen-thawed embryos between blastocyst developmental stages (early vs mid, P<0.05; mid vs expanded, P<0.01; early vs expanded, P<0.001). The post-thawing survival rate of blastocysts frozen at Day 7 (62%) of culture was higher compared with that of Day 8 (45%), but there was no difference in survival rate between Day 7 and 8 of culture. The results indicate that the quality and developmental stage of blastocysts are important factors influencing their survival after freezing and thawing.

Journal Article↗

PubMed1998

Implications of a developmental-stage-dependent thylakoid-bound protease in the stabilization of the light-harvesting pigment-protein complex serving photosystem II during thylakoid biogenesis in red kidney bean

Intact etioplasts of bean (Phaseolus vulgaris) plants exhibit proteolytic activity against the exogenously added apoprotein of the light-harvesting pigment-protein complex serving photosystem II (LHCII) that increases as etiolation is prolonged. The activity increases in the membrane fraction but not in the stroma, where it remains low and constant and is mainly directed against LHCII and protochlorophyllide oxidoreductase. The thylakoid proteolytic activity, which is low in etioplasts of 6-d-old etiolated plants, increases in plants pretreated with a pulse of light or exposed to intermittent-light (ImL) cycles, but decreases during prolonged exposure to continuous light, coincident with chlorophyll (Chl) accumulation. To distinguish between the control of Chl and/or development on proteolytic activity, we used plants exposed to ImL cycles of varying dark-phase durations. In ImL plants exposed to an equal number of ImL cycles with short or long dark intervals (i.e. equal Chl accumulation but different developmental stage) proteolytic activity increased with the duration of the dark phase. In plants exposed to ImL for equal durations to such light-dark cycles (i.e. different Chl accumulation but same developmental stage) the proteolytic activity was similar. These results suggest that the protease, which is free to act under limited Chl accumulation, is dependent on the developmental stage of the chloroplast, and give a clue as to why plants in ImL with short dark intervals contain LHCII, whereas those with long dark intervals possess only photosystem-unit cores and lack LHCII.

Journal Article↗

PubMed1990

Neurotoxicity of kainic acid in the rat cochlea during early developmental stages.

The neurotoxic effect of kainic acid (KA) was investigated by electron microscopy in rat cochleas at two developmental stages: 17 days of gestation (17 G) and postnatal day 1 (PN 1). In each animal, one cochlea was injected with 1 nmol KA diluted into 2 ml artificial perilymph, while the other cochlea was only injected with artificial perilymph as a control. Ten minutes later, the cochleas were perfused with fixative, removed and processed for electron microscopy. The KA injection resulted in marked swelling of the majority of afferent fibers, i.e. the peripheral processes of spiral ganglion neurons. In the 17 G cochlea, swollen fibers were traced from the perikarya to the undifferentiated otocyst epithelium. Following birth, swollen afferents in the PN 1 cochlea were in contact with both inner (IHCs) and outer hair cells (OHCs), which were now differentiated. At both stages of development, a subclass of small afferent nerves were unaffected. At PN 1, the KA-insensitive afferents only contacted the OHCs. These fibers probably belong to the spiral system of afferents and are related to type II spiral ganglion cells. Conversely, KA-sensitive afferents probably belong to the radial system, related to type I spiral ganglion cells. This system is specific for IHCs in adult cochleas and appears to innervate both IHCs and OHCs at early developmental stages. These findings also indicate that KA neurotoxicity appears very early in the cochlea, at a prenatal time (17 G) before the presynaptic partners of afferent terminals (namely the IHCs) are differentiated.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMedDate not supplied

Transcriptional potentials of the beta-like globin genes at different developmental stages in transgenic mice and hemoglobin switching.

Developmental-stage-specific regulation and physiological levels of expression of the globin genes can be recaptured in transgenic mice carrying a YAC/BAC- or cosmid-based construct. By contrast, proper developmental regulation and high-level expression cannot be achieved coordinately in transgenic mice carrying a more manipulated construct, such as a plasmid-based globin gene construct. These differences provide us an opportunity to define the requirements for a developmentally regulated, high-level expression of the globin genes in vivo. To achieve this, as a first step, we studied maximum transcriptional potentials of the beta-globin genes at various stages of development. microLCR-enhanced expression of the epsilon-, gamma-, and beta-globin genes driven by their minimal promoters was estimated and compared with that in betaYAC transgenic mice. Quantitative measurements of steady state mRNA levels of the epsilon-, gamma-, and beta-globin genes showed that the microLCR was able to enhance expression of each beta-like globin gene to levels similar to those in the betaYAC mice. Moreover, transcriptional potentials of each globin gene were unchanged during the entire course of development. These observations indicate that the highest level of expression of the globin genes can be achieved in both embryonic and definitive erythropoiesis regardless of developmental specificity of the genes. This finding implies that transcription suppression is the major mechanism of the developmental specificity of the expression of the beta-like globin genes.

Animals↗

PubMedDate not supplied

[The effect of betain on biology and morphology of developmental stages of Eimeria acervulina in broiler chicks experimentally infected].

Purposing to investigate the betaine effect on biology and morphology of developmental stages of Eimeria acervulina, 420 broiler chicks Cobb were experimentally inoculated with 2 x 10(5) sporulated oocysts and housed in battery cages in a block design with five treatments and six replicates each, including a positive control, a group treated with salinomycin and growth promoter plus three levels of betaine as additive in the feed at 0.05, 0.10 and 0.15%. Measurements of oocysts, sporocysts and endogenous stages were performed as morphological parameters, while pre patent and patent periods and sporulation time were taken as biological parameters. Morphology was also associated with the mathematical constant Phi (1.618) to evaluate possible relationship. Betaine was able to cause modifications in both biology and morphology of oocysts and sporocysts, whereas it was weakly able to affect developmental stages based on trophozoites and macrogamonts measurements. According to the measures of sporocysts E. acervulina development was closely related to Phi.

Animals↗

PubMed2001

Actin cytoskeleton reorganization of the apoptotic nurse cells during the late developmental stages of oogenesis in Dacus oleae.

In the present study, we demonstrate the actin cytoskeleton reorganization during nurse cells apoptosis of the olive fruit fly Dacus oleae. At the developmental stage 9A of oogenesis, the actin microfilaments are assembled in numerous ring canals and subcortically support all the nurse cells, as is shown by phalloidin-FITC staining. During the following stages, 9B and 10A, this structural pattern remains the same. The developmental stage 10B is characterized by actin microfilament rearrangement and formation of actin cables that are symmetrically organized around the nurse cell nuclei. At stage 11, when the dumping process begins, these actin cables seem to retain each nurse cell nucleus in the cell center, away from blocking the ring canals. The early stage 12 is characterized by an asynchronous nurse cell nuclear chromatin condensation, while at late stage 12 the actin cables become very thick, as adjacent ones overlap one another and traverse the disorganized apoptotic nurse cell nuclei that already have fragmented DNA, as is demonstrated by acridine orange staining and TUNEL assay. Finally, during stage 13, the apoptotic nuclear remnants are phagocytosed by the neighboring follicle cells. The data presented herein compared to previous reported results in Drosophila [Nezis et al., 2000: Eur J Cell Biol 79:610-620], demonstrate that actin cytoskeleton reorganization during nurse cell apoptosis is a developmentally regulated physiological mechanism, phylogenetically conserved in higher Dipteran.

Acridine Orange↗

PubMed1984

Different aggregation properties of sea urchin embryonic cells at different developmental stages. I. Stage specificity of aggregation factors solubilized by butanol.

Surface proteins solubilized with butanol from purified plasma membranes of sea urchin embryos at different developmental stages were tested for their aggregation promoting activity on dissociated cells. Cells used for the assays were obtained either from blastulae or from embryos at the 16 cell stage. Results show that a strong enhancement of cell aggregation was produced only when extracted proteins and dissociated cells were obtained from embryos at the same developmental stage.

Animals↗

PubMed1997

Evaluation of cDNA libraries from different developmental stages of Schistosoma mansoni for production of expressed sequence tags (ESTs).

A comparative study of the gene expression profile in different developmental stages of Schistosoma mansoni has been initiated based on the expressed sequence tag (EST) approach. A total of 1401 ESTs were generated from seven different cDNA libraries constructed from four distinct stages of the parasite life cycle. The libraries were first evaluated for their quality for a large-scale cDNA sequencing program. Most of them were shown to have less than 20% useless clones and more than 50% new genes. The redundancy of each library was also analyzed, showing that one adult worm cDNA library was composed of a small number of highly frequent genes. When comparing ESTs from distinct libraries, we could detect that most genes were present only in a single library, but others were expressed in more than one developmental stage and may represent housekeeping genes in the parasite. When considering only once the genes present in more than one library, a total of 466 unique genes were obtained, corresponding to 427 new S. mansoni genes. From the total of unique genes, 20.2% were identified based on homology with genes from other organisms, 8.3% matched S. mansoni characterized genes and 71.5% represent unknown genes.

Animals↗

PubMed1998

Effect of developmental stage on bovine oocyte plasma membrane water and cryoprotectant permeability characteristics.

Knowledge of bovine oocyte plasma membrane permeability characteristics at different developmental stages in the presence of cryoprotective agents (CPAs) is limited. The objective of this study was to determine the oolema hydraulic conductivity (Lp), cryoprotectant permeability (P[CPA]), and reflection coefficient (sigma) for immature (germinal vesicle stage, GV) and in vitro-matured (metaphase II, MII) bovine oocytes. Two commonly used cryoprotective agents, dimethyl sulfoxide (DMSO) and ethylene glycol (EG), were studied. Osmometric studies were performed using a micromanipulator connected to an inverted microscope at 22 +/- 2 degrees C. Each oocyte was immobilized via a holding pipette, and osmotically induced volume changes over time (dv/dt) were recorded. The Lp values for GV and MII oocytes in DMSO (L(p)DMSO) were 0.70 +/- 0.06 and 1.14 +/- 0.07 microm/min/atm (mean +/- SEM) and in EG (L(p)EG) were 0.50 +/- 0.06 and 0.83 +/- 0.07 microm/min/atm, respectively. Estimates of P(DMSO) for GV and MII oocytes were 0.36 +/- 0.03 and 0.48 +/- 0.03 microm/sec, and PEG values for GV and MII oocytes were 0.22 +/- 0.03, 0.37 +/- 0.03 microm/sec, respectively. The values for GV and MII oocytes in DMSO (sigma[DMSO]) were 0.86 +/- 0.03 and 0.90 +/- 0.04 and in EG (sigma[EG]) were 0.94 +/- 0.03 and 0.76 +/- 0.04, respectively. These data demonstrate that bovine oolema permeability coefficients to water and cryoprotectants change after in vitro maturation. Furthermore, the bovine oocyte P(DMSO) is higher than the P(EG). These results may provide a biophysical basis for developing criteria for choosing optimal CPAs and for minimizing damage during addition and removal of the CPAs. Additionally, these data support the hypothesis that different procedures may be required for optimal cryopreservation of different oocyte developmental stages.

Animals↗