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Progestogen-dependent endometrial protein in women: tissue concentration in relation to developmental stage and to serum hormone levels.

The presence of a progestogen-dependent endometrial protein (PEP) in women was previously described. The present study was undertaken to determine whether the occurrence or the concentration of PEP in endometria is related to the developmental stage of tissues and/or to the levels of endogenous progesterone (P) or 17 beta-estradiol (E) to which the tissues are exposed. Samples of serum and endometrial tissue in different developmental stages were obtained from pregnant women and women with normal menstrual cycles and were analyzed for PEP. In addition, the sera were analyzed for P and E. The occurrence of PEP in the endometrium of all of the women who were studied was found to be correlated with the heightened secretory activity and with the decidual change in the tissue. Furthermore, in women with normal cycles, the detection of PEP was associated with high serum levels of P. Comparison of serum levels of P with endometrial levels of PEP in pregnant women and in women in the mid-secretory phase of the menstrual cycle indicated that a 2.8-fold rise in serum levels of P during pregnancy was accompanied by a comparable (about threefold) rise in tissue levels of PEP. Therefore, we conclude that, in pregnant women and women with normal cycles, the levels of PEP in the endometrium reflect developmental changes within the endometrium, and that those levels are directly related to serum levels of P.

Adult↗

Differential expression of two sporulation specific sigma factors of Streptomyces aureofaciens correlates with the developmental stage.

In previous experiments, the Streptomyces aureofaciens (Sa) rpoZ, and sigF genes have been shown to encode putative sigma factors essential in differentiation. In an attempt to investigate expression of these genes during differentiation, we have performed S1 nuclease mapping using RNA prepared from Sa in various developmental stages. Two putative promoters were identified upstream of the rpoZ coding region. The promoters significantly differed in their strength, and were active in distinct developmental stages; the weaker, rpoZ-P1, was active only in substrate mycelium, and the stronger, rpoZ-P2, was induced at the beginning of aerial mycelium formation. Transcriptional analysis of sigF revealed two apparent transcription start points, both being detectable only in the late phase of aerial mycelium formation. No sigF transcription was detected in an rpoZ-disrupted Sa strain. Promoter-bearing DNA fragments from rpoZ and sigF were inserted into several promoter-probe vectors, to give expression patterns consistent with the results of direct RNA analysis. The results implicate temporally different expression of rpoZ and sigF during the differentiation of Sa, and direct or indirect dependence of sigF expression on the rpoZ-encoded putative sigma factor, thus indicating a cascade of sigma factors in Streptomyces development.

Amino Acid Sequence↗

Identification of a novel developmental stage marking lineage commitment of progenitor thymocytes.

Bipotent progenitors for T and natural killer (NK) lymphocytes are thought to exist among early precursor thymocytes. The identification and functional properties of such a progenitor population remain undefined. We report the identification of a novel developmental stage during fetal thymic ontogeny that delineates a population of T/NK-committed progenitors (NK1. 1(+)/CD117(+)/CD44(+)/CD25(-)). Thymocytes at this stage in development are phenotypically and functionally distinguishable from the pool of multipotent lymphoid-restricted (B, T, and NK) precursor thymocytes. Exposure of multipotent precursor thymocytes or fetal liver- derived hematopoietic progenitors to thymic stroma induces differentiation to the bipotent developmental stage. Continued exposure to a thymic microenvironment results in predominant commitment to the T cell lineage, whereas coculture with a bone marrow-derived stromal cell line results in the generation of mature NK cells. Thus, the restriction point to T and NK lymphocyte destinies from a multipotent progenitor stage is marked by a thymus-induced differentiation step.

Animals↗

Characterization of mouse monoclonal antibodies against common and repeated epitopes of different developmental stages of Schistosoma mansoni.

Eight IgM monoclonal antibodies (mAbs) obtained from a Schistosoma mansoni chronically infected mouse immunized twice with adult worms butanolic extract (BE) and soluble egg antigen (SEA) were characterized by immunochemical methods. An intense cross-reactivity between different developmental stages was observed by indirect immunofluorescence assay (IIFA) with five anti-SEA mAbs. These mAbs appeared to recognize glycosidic residues, as suggested by 1) the inhibition of their reactivity by periodate oxidation of SEA, 2) the multiple polypeptide recognition in radioimmunoprecipitation and immunoblot assays and 3) reactivity with KLH. Anti-SEA mAbs were able to mediate in vitro killing of schistosomula but they were not consistently able to mediate passive transfer immunity in vivo. Three of anti-SEA mAbs were suitable for the performance of a sandwich ELISA for antibody detection in S. mansoni infected patients, allowing an increase in the signal to noise ratio as compared to the direct ELISA SEA method. Three anti-BE mAbs showed a more stage restricted pattern of antigen recognition by IIFA. Only one out of three seemed to be directed against glycan residues, but the other mAbs showed a plural pattern of polypeptide recognition on BE immunoblot, suggesting that repeated epitopic motifs are also present in different proteins within the same parasite developmental stage.

Animals↗

The development of the chamber angle in the rat eye. Morphological characteristics of developmental stages.

The pre- and postnatal development of the chamber angle region was studied in the rat. The eyes of five weight-matched littermates were investigated by light- and electron microscopy at each of the following times: prenatal days 14, 17, 18, 21; day of birth; postnatal days 5, 10, 15, 20, 40, 60, 100, 200. Major developmental stages are demonstrated and the morphological characteristics of each stage shown. Developmental periods in the rat are correlated with periods that have previously been outlined in the human (Remé and Lalive 1981). Major periods comprise the following events: I. anlage formation (day 17 prenatal - day postnatal); II. differentiation into definitive structures (days 5-10); III. specialization of definitive structures (days 10-20); IV. final components (days 20-60); V. final molding and rarefaction (days 60-100). Our data indicate that the developing rat eye is a suitable experimental model for studies on the development of the chamber angle. Moreover, this study provides baseline data for a separate investigation on morphogenetic events contributing to the remodeling processes occurring within the vertebrate chamber angle.

Animals↗

Fractionation of mouse malarious blood according to parasite developmental stage, using a Percoll-sorbitol gradient.

Asexual intraerythrocytic malarial parasites permeabilize the membrane of their host cell to small monelectrolytes and anions. Since permeabilization increases with parasite maturation, this property has been used previously to fractionate blood infected with Plasmodium falciparum and P. knowlesi according to the developmental stage of the parasite, using Percoll-sorbitol density gradients. We have extended this method to fractionate mouse blood infected with four species of rodent malaria: P. chahaudi, P. vinckei, P. voelii and P. berghei. While the method works in principle in this case, the polyparasitism which characterizes these species prevented explicit separation according to developmental stage. Hence, erythrocytes harbouring several ring-stage parasites appeared in the same fraction which contained cells hosting a single trophozoite, and polyparasitized trophozoites were associated with singly-infected schizont. This observation implies that permeabilization of the host cell membrane results from the integrated metabolic activity of the parasite(s) and is not related to a specific phase of parasite development.

Animals↗

Neocortical injuries at different developmental stages determine different susceptibility to seizures induced in adulthood.

Gliosis, axonal sprouting and remodelling of nerve connections in the injured brain have been regarded as epileptogenic processes dependent on the age when the injury was inflicted. The present study examines whether brains injured at different developmental stages may acquire different susceptibility to experimental status epilepticus induced in adulthood. In 6- and 30-day-old Wistar rats (P6s and P30s, respectively), a mechanical injury was performed in the left cerebral hemisphere. On postnatal day 60, all the animals and controls received single injections of kainic acid to evoke status epilepticus. During a 6-h period following the injection, the animals were observed continuously and motor symptoms of seizure activity were recorded and rated. P6s showed significantly lower intensity of kainic acid-induced epileptic symptoms and significantly longer survival than controls or P30s. In P30s, no significant change was detected. The data provide evidence that the developmental stage when the brain is injured determines epileptogenecity of the lesion. However, a considerable discrepancy between these data and those obtained previously following pilocarpine administration in the same experimental models of brain injury shows that each of the two models of epilepsy may display different aspects of the same age-dependent process triggered by the brain injury.

Animals↗

Two different cDNAs code for L-DOPA decarboxylase in the white prepuparium and eclosion developmental stages of the insect Ceratitis capitata1.

In insects the DOPA decarboxylase gene (Ddc) is expressed in the epidermis where dopamine derivatives promote cuticle sclerotization and the central nervous system participating in the synthesis of serotonin and dopamine. The temporal course of Ddc enzyme activity during the white prepuparium and eclosion developmental stages primarily reflects this role. Here we report that a cDNA clone for l-DOPA decarboxylase (EC 4.1.1.28) has been isolated from the white puparium stage of the insect Ceratitis capitata, the cDNA sequence has been determined and it corresponds to an mRNA of 1341 base pairs. The comparison of this cDNA sequence with that of the eclosion stage which has been previously isolated in our laboratory revealed that the cDNA from the eclosion stage codes for protein consisting of 431 amino acid (aa) residues whereas that from the white prepuparium stage for protein of 425 aa residues. In addition there is a region of 33 aa residues close to the N-terminal region of the deduced protein in which marked differences in the aa composition are observed. This result was further confirmed by Northern blot analysis in which mRNAs isolated from both developmental stages were only hybridized with their respective polymerase chain reaction synthesized stage specific probes. These results suggest that during development of the insect C. capitata two different mRNAs are produced at the white prepuparium and the eclosion stages, giving rise to two protein isoforms presumably in response to different signals.

Amino Acid Sequence↗

Clinical experiences of ICSI-ET thawing cycles with embryos cryopreserved at different developmental stages.

PURPOSE: The aim of our study was to analyze factors including survival, implantation and pregnancy rate, patients' age and BMI, abortions and extra uterine pregnancies that might influence the outcome of ICSI-ET thawing cycles. METHODS: A total of 147 cycles with embryos cryopreserved at different developmental stages were retrospectively evaluated. RESULTS: No difference was found in the survival, implantation and pregnancy rates of embryos cryopreserved on Day 2-3 and 5. However, in the pregnant group significantly higher implantation rate was observed with Day 5 blastocysts then with Day 2 or 3 early embryos. We found no difference in the number of abortions and extra uterine pregnancies between fresh and frozen ICSI-ET cycles. Higher BMI was found in the pregnant than in the nonpregnant group. However, the age of patient had no effect on the results. CONCLUSIONS: Developmental stage of embryo and patients' BMI influences the success of ICSI-ET thawing cycles.

Body Mass Index↗

Protein composition of various developmental stages of dipetalonema viteae (Filarioidea).

The protein composition of various developmental stages of Dipetalonema viteae was analysed on polyacrylamide slab gels in the presence of sodium-dodecylsulphate. When the total proteins of adult male and female parasites, microfilariae, eggs, and third-stage larvae were compared, apparent qualitative similarities between mature and immature filariae were observed. However, several stage specific components were also identified.

Animals↗

Immunogenicity of homogenates of the developmental stages of Litomosoides carinii in albino rats.

Homogenates of different developmental stages of the filarial parasites, L. carinii, emulsified in Freund's complete adjuvant have been examined for their efficacy in conferring immunity to the infection in the albino rats. The results revealed that sonicated preparations of microfilariae and infective larvae induce high resistance to the infection in the animals. In contrast, soluble antigens of adults and sonicated homogenates of adult males were ineffective to induce such resistance. Some resistance seen when sonicated adult worms of both sexes were used as immunogens appears to be due to the microfilarial antigen present in the extracts.

Animals↗

S100B expression defines a state in which GFAP-expressing cells lose their neural stem cell potential and acquire a more mature developmental stage.

During the postnatal development, astrocytic cells in the neocortex progressively lose their neural stem cell (NSC) potential, whereas this peculiar attribute is preserved in the adult subventricular zone (SVZ). To understand this fundamental difference, many reports suggest that adult subventricular GFAP-expressing cells might be maintained in immature developmental stage. Here, we show that S100B, a marker of glial cells, is absent from GFAP-expressing cells of the SVZ and that its onset of expression characterizes a terminal maturation stage of cortical astrocytic cells. Nevertheless, when cultured in vitro, SVZ astrocytic cells developed as S100B expressing cells, as do cortical astrocytic cells, suggesting that SVZ microenvironment represses S100B expression. Using transgenic s100b-EGFP cells, we then demonstrated that S100B expression coincides with the loss of neurosphere forming abilities of GFAP expressing cells. By doing grafting experiments with cells derived from beta-actin-GFP mice, we next found that S100B expression in astrocytic cells is repressed in the SVZ, but not in the striatal parenchyma. Furthermore, we showed that treatment with epidermal growth factor represses S100B expression in GFAP-expressing cells in vitro as well as in vivo. Altogether, our results indicate that the S100B expression defines a late developmental stage after which GFAP-expressing cells lose their NSC potential and suggest that S100B expression is repressed by adult SVZ microenvironment.

Animals↗

Gene expression profiling in porcine mammary gland during lactation and identification of breed- and developmental-stage-specific genes.

A total of 28941 ESTs were sequenced from five 5'-directed non-normalized cDNA libraries, which were assembled into 2212 contigs and 5642 singlets using CAP3. These sequences were annotated and clustered into 6857 unique genes, 2072 of which having no functional annotations were considered as novel genes. These genes were further classified into Gene Ontology categories. By comparing the expression profiles, we identified some breed- and developmental-stage-specific gene groups. These genes may be relative to reproductive performance or play important roles in milk synthesis, secretion and mammary involution. The unknown EST sequences and expression profiles at different developmental stages and breeds are very important resources for further research.

Animals↗

Chilling sensitivity of carp (Cyprinus carpio) embryos at different developmental stages in the presence or absence of cryoprotectants: work in progress.

The unsolved problem of cryopreservation of the yolk-rich teleost embryos may be related, in part, to their sensitivity to chilling and cryoprotective agents. The aim of this study was to gain data on the sensitivity of carp embryos to low temperatures at different developmental stages and on the possible protective and toxic effects of cryoprotectants. A total of 86,400 morulae, half-epiboly and heartbeat-stage embryos was selected and then placed in water or in 1 M methanol, dimethyl sulfoxide (Me2SO), glycerol or 0.1 M sucrose solution at 0, 4 or 24 degrees C for 5 min or 1 h. Following these treatments, the embryos were held in a 24 degrees C water bath until the evaluation of hatching rates. In every developmental stage a significant decrease of hatching rates following exposure to 4 or 0 degree C was detected. Sensitivity to chilling changed significantly with development (heartbeat < morula < half-epiboly). Half-epiboly stage embryos were less sensitive to a short period of exposure to cryoprotectants than morula and heartbeat stages. A 1-h exposure to cryoprotectants revealed a stage dependent sensitivity. Toxicity increased in the order of methanol < Me2SO < glycerol in morula and half-epiboly stages, and methanol < glycerol < Me2SO in the heartbeat stage. The results show morulae are partially protected against chilling in Me2SO and sucrose, half-epiboly in Me2SO, sucrose and methanol, and heartbeat-stage in methanol and glycerol. The results further suggest that carp embryos are sensitive to chilling and that toxicity and protective effects against chilling of cryoprotectants are stage-dependent. The finding on the low chilling sensitivity of heartbeat-stage embryos and the protective effect of certain cryoprotectants may be useful in designing cryopreservation protocols.

Animals↗

Different neuroligands and signal transduction pathways stimulate CREB phosphorylation at specific developmental stages along oligodendrocyte differentiation.

We have shown previously that the pattern of expression of the transcription factor CREB (cyclic AMP-response element binding protein) in developing oligodendrocytes (OLGs) suggests a role during a period that precedes the peak of myelination in rat brain. We have now investigated the signaling pathways that could be responsible for activating CREB by phosphorylation at different stages along OLG maturation. CREB phosphorylation was studied in short-term cultures of immature OLG precursor cells and young OLGs isolated from 4- and 11-day-old rat cerebrum, respectively. The results indicated that at both developmental stages, CREB phosphorylation could be stimulated by either increased concentrations of cyclic AMP and cyclic AMP-dependent protein kinase activation or increased Ca2+ levels and a protein kinase C activity. The results also showed that CREB phosphorylation in immature OLG precursor cells could be up-regulated by treatment with histamine, carbachol, glutamate, and ATP (neuroligands known to increase Ca2+ levels in these cells), by signaling cascade(s) that involve a protein kinase C activity, as well as the mitogen-activated protein kinase pathway. In contrast, in cells isolated from 11-day-old rats, at a developmental stage that immediately precedes the beginning of the active period of myelin synthesis, CREB phosphorylation was only stimulated by treatment with the beta-adrenergic agonist isoproterenol in a process that appears to be mediated by a cyclic AMP/cyclic AMP-dependent protein kinase-dependent pathway. These results support the idea that CREB could be a mediator of neuronal signals that, coupled to specific signal transduction cascades, may play different regulatory roles at specific stages along OLG differentiation.

Adenosine Triphosphate↗

Gene expression profiling in human fetal liver and identification of tissue- and developmental-stage-specific genes through compiled expression profiles and efficient cloning of full-length cDNAs.

Fetal liver intriguingly consists of hepatic parenchymal cells and hematopoietic stem/progenitor cells. Human fetal liver aged 22 wk of gestation (HFL22w) corresponds to the turning point between immigration and emigration of the hematopoietic system. To gain further molecular insight into its developmental and functional characteristics, HFL22w was studied by generating expressed sequence tags (ESTs) and by analyzing the compiled expression profiles of liver at different developmental stages. A total of 13,077 ESTs were sequenced from a 3'-directed cDNA library of HFL22w, and classified as follows: 5819 (44.5%) matched to known genes; 5460 (41.8%) exhibited no significant homology to known genes; and the remaining 1798 (13.7%) were genomic sequences of unknown function, mitochondrial genomic sequences, or repetitive sequences. Integration of ESTs of known human genes generated a profile including 1660 genes that could be divided into 15 gene categories according to their functions. Genes related to general housekeeping, ESTs associated with hematopoiesis, and liver-specific genes were highly expressed. Genes for signal transduction and those associated with diseases, abnormalities, or transcription regulation were also noticeably active. By comparing the expression profiles, we identified six gene groups that were associated with different developmental stages of human fetal liver, tumorigenesis, different physiological functions of Itoh cells against the other types of hepatic cells, and fetal hematopoiesis. The gene expression profile therefore reflected the unique functional characteristics of HFL22w remarkably. Meanwhile, 110 full-length cDNAs of novel genes were cloned and sequenced. These novel genes might contribute to our understanding of the unique functional characteristics of the human fetal liver at 22 wk.

Cloning, Molecular↗

Ontogeny of the invertebrate humoral immune response: studies on various developmental stages of the American cockroach (Periplaneta americana).

Earlier studies revealed that a specific adaptive humoral immune response can be induced in the American cockroach (Periplaneta americana) to the soluble protein complex, Honeybee venom (HBT). We have undertaken a series of ontogenetic studies to determine if there are differences between the protective responses of roaches representing different developmental stages. Our results indicated that the response to HBT in the immature cockroach (nymphs weighing 200-500 mg) was characterized by a significant lag period before immune protection began to develop. However, by the second week of the response, reactivity was comparable to that of the adult. Old adult male roaches (animals 5 months into adulthood) displayed a significant decline in reactivity during the early phases of the response in comparison to younger adults, and in general, appeared to be less vigorous in generating protection. Both the nymph and old adult roaches demonstrated good secondary responsiveness. The results from these experiments indicated that the developmental stage of the roach could be directly correlated to the degree of immunocompetency possessed by the animal. This is similar to the ontogenetic sequence typically found for immune reactivity in vertebrates.

Age Factors↗

Immunological analysis of cells of Nocardia at various developmental stages during morphogenesis.

High antibody titres and a single precipitin line of identity were obtained with hyperimmune rabbit serum prepared against filamentous cells of Nocardia asteroides strain 55 and the cells of three other strains of Nocardia, indicating the presence of a group antigen between the strains. High titres were also obtained with the hyperimmune serum and the three different cell forms (filamentous, rod-like and spherical cells) of the homologous strain, indicating that each developmental stage was highly antigenic. The antibodies were completely removed by absorption with the various cell forms, suggesting a similarity of the antigens. A single common antigen was demonstrated between the three developmental stages by immunodiffusion, indicating that there was no antigenic alteration occuring during morphogenesis of the genus Nocardia.

Agglutination Tests↗