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PubMed1993

Hemicerebellectomy and motor behaviour in rats. III. Kinematics of recovered spontaneous locomotion after lesions at different developmental stages.

The locomotion of rats with a right hemicerebellectomy (HCb) performed in adulthood was compared by means of kinematic analysis with the locomotion of rats with a similar lesion performed on the first postnatal day. The age at which the animals received cerebellar lesion made a significant difference with respect to the locomotor strategies utilized in adulthood. During stance, neonatal operate rats showed a clear hyperextension of both hindlimbs but not of the forelimbs. Their locomotor posture was characterized by spinal flexion with the head held lower than normal. During swing, they showed a tendency towards 'high stepping'. Their steps were regular and symmetrical but hypometric. Adult lesioned animals displayed a marked extensor hypotonia, ipsilateral to the lesion during stance and a relevant hyperflexion affecting both sides, during swing. Alteration of the interlimb coordination and modified sequence of steps were also observed. Thus, a highly asymmetrical, impaired and unstable locomotion was displayed by this group of animals. The present findings demonstrate the importance of the age-at-lesion factor in determining the motor strategies in the recovery of locomotor function after HCb in the rat. This evidence is discussed in the light of the widespread anatomical remodelling already demonstrated following neonatal, but not adult, HCb in rats.

Aging↗
PubMedDate not supplied

[Laboratory investigation of the chemical disinfection against different developmental stages of the housefly (Musca domestica Linnaeus, 1758)].

Possibilities for disinfection of the developmental stages from the housefly (Musca domestica) were investigated under laboratory conditions. The developmental stages (eggs, larvae I, II, III, pupae and adults) were sprayed with solutions of disinfectants on the basis of p-chlorine-m-cresol and o-phenylphenol at different concentrations (0.025%-3%). The effectiveness of both disinfectants was established by determining the emergence rate of eggs and pupae and of the live evidence of larvae and adults. Transmission electron microscopic investigations of the cover of the eggs revealed the bactericidal effect of both disinfectants. These results show the necessity of extending its use also to noxious arthropods.

Animals↗
PubMed1978

Observations on living rat spermatogenic cells in different developmental stages.

In the seminiferous tubules of the rat, as in most mammalian species, the developing germ cells form associations with constant cell composition. These cellular associations or stages follow each other in a regular manner along the seminiferous tubules giving rise to seminiferous epithelial wave. When a freshly isolated unstained seminiferous tubulus of the rat is subjected to transillumination under a stereomicroscope, the different segments of the seminiferous epithelial wave absorb light in a characteristic manner permitting their recognition. Using this technique, small segments with accurately known cell composition can be isolated and studied in living state with phase-contrast microscopy. In several cases, the phase-contrast microscopy gives more information about the cell morphology than conventional histological methods. In this study all major developmental steps from early spermatogonia to mature spermatids have been described. The findings of the present study can be used as reference material in the evaluation and identification of the various cell types of the seminiferous tubules obtained, e.g. by the Staput fractionation method. In addition, the findings may be helpful in the evaluation of spermatogenic and Sertoli cells in culture conditions.

Animals↗
PubMed1993

Partial characterization of proteolytic enzymes in different developmental stages of Ostertagia ostertagi.

Proteolytic enzymes present in extracts of third (L3) and fourth (L4) stage larvae and adults of the cattle nematode Ostertagia ostertagi were defined on the basis of pH optima and proteinase inhibitor sensitivity in spectrophotometric assays using azocasein and elastin-orcein as protein substrates. Evidence that different classes of proteinases are expressed in a stage specific manner was provided by the contrasting pH optima and inhibitor sensitivities shown by the enzymes in the different parasite stages. Stage specificity was confirmed by gelatin-substrate analysis. In addition, proteolytic activity was sought in the excretory/secretory products (ES) of the L4 following simple in vitro culture. Contrasting pH and inhibitor sensitivities as well as gelatin-substrate analysis showed that different proteinases were present in somatic L4 extracts and L4 ES products. The secreted proteinases may be useful targets for serodiagnosis or vaccination.

Animals↗
PubMed1987

Localization and quantitation of 125I-epidermal growth factor binding in mouse embryonic tooth and other embryonic tissues at different developmental stages.

We have shown earlier that epidermal growth factor (EGF) inhibits morphogenesis and cell differentiation in mouse embryonic teeth in organ culture. This inhibition depends on the stage of tooth development so that only teeth at early developmental stages respond to EGF (A-M. Partanen, P. Ekblom, and I. Thesleff (1985) Dev. Biol. 111, 84-94). We have now studied the quantity and pattern of EGF binding in teeth at various stages of development by incubating the dissected tooth germs with 125I-labeled EGF. Although the quantity of 125I-EGF binding per microgram DNA stays at the same level, localization of 125I-EGF binding by autoradiography reveals that the distribution of binding sites changes dramatically. In bud stage the epithelial tooth bud that is intruding into the underlying mesenchyme has binding sites for EGF, but the condensation of dental mesenchymal cells around the bud does not bind EGF. At the cap stage of development the dental mesenchyme binds EGF, but the dental epithelium shows no binding. This indicates that the dental mesenchyme is the primary target tissue for the inhibitory effect of EGF on tooth morphogenesis during early cap stage. During advanced morphogenesis the binding sites of EGF disappear also from the dental papilla mesenchyme, but the dental follicle which consists of condensed mesenchymal cells surrounding the tooth germ, binds EGF abundantly. We have also studied EGF binding during the development of other embryonic organs, kidney, salivary gland, lung, and skin, which are all formed by mesenchymal and epithelial components. The patterns of EGF binding in various tissues suggest that EGF may have a role in the organogenesis of epitheliomesenchymal organs as a stimulator of epithelial proliferation during initial epithelial bud formation and branching morphogenesis. The results of this study indicate that EGF stimulates or maintains proliferation of undifferentiated cells during embryonic development and that the expression of EGF receptors in different organs is not related to the age of the embryo, but is specific to the developmental stage of each organ.

Animals↗
PubMed1998

Variation in acid hydrolase enzyme titers in different developmental stages of Aedes togoi.

Genetic mechanisms of filarial nematode susceptibility were studied in Aedes togoi. Acid hydrolases may play an important role in this process, including humoral or cell-mediated defenses. Levels of acid phosphatase, alpha-glucosidase, beta-glucuronidase, and N-acetyl-beta-glucosaminidase were determined for 1st- and 4th-instar larvae, male and female pupae, and 1- and 7-day-old adults using fluorometric and colorimetric assays. Acid phosphatase activity was highest in 1-day-old adults, moderate in larvae and pupae, and lowest in 7-day-old adults. Female 7-day-old adults had significantly higher levels than males of the same age. Moderate levels of alpha-glucosidase were found in larvae, with progressive increases in activity from pupae to 7-day-old adults. Levels in male pupae and 1-day-old males were higher than in females, but activity was twice as high in 7-day-old females. Activity of beta-glucuronidase was greater in adults, with females showing a 2-fold higher level than males at 7 days. In contrast, N-acetyl-beta-glucosaminidase activity was highest in 1st- and 4th-instar larvae and 1-day-old males and females. Activity also was significantly higher in male pupae, slightly greater in 1-day-old males, but twice as high in 7-day-old females when compared to males of the same age. Results showed significant changes and variation in acid hydrolase enzyme titers in the different life stages of Ae. togoi. These and other specific acid hydrolases could prove effective in monitoring biochemical and genetic changes in mosquito populations.

Aedes↗
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↗
PubMed1998

Cytokines promote the survival of mouse cranial sensory neurones at different developmental stages.

To investigate when the neurotrophic cytokines ciliary neurotrophic factor (CNTF), leukaemia inhibitory factor (LIF), oncostatin-M (OSM), interleukin-6 (IL-6) and cardiotrophin-1 (CT-1) act on developing sensory neurones and whether they co-operate with neurotrophins in regulating neuronal survival, we studied the in vitro trophic effects of these factors on two well-characterized populations of cranial sensory neurones at closely staged intervals throughout embryonic development. The cutaneous sensory neurones of the trigeminal ganglion, which show an early, transient survival response to BDNF and NT3 before becoming NGF-dependent, were supported by CNTF, LIF, OSM and CT-1 during the late fetal period, several days after the neurones become NGF-dependent. At this stage of development, these cytokines promoted the survival of a subset of NGF-responsive neurones. The enteroceptive neurones of the nodose ganglion, which retain dependence on BDNF throughout fetal development, were supported throughout their development by CNTF, LIF, OSM and CT-1, and displayed an additional survival response to IL-6 in the late fetal period. These findings indicate that populations of sensory neurones display different developmental patterns of cytokine responsiveness and show that embryonic trigeminal neurones pass through several phases of differing neurotrophic factor survival requirements.

Aging↗
PubMed1996

Survival rates and sex ratio of bovine IVE embryos frozen at different developmental stages on day 7.

Two experiments were designed to determine the effects of stage of development on Day 7 of in vitro-produced bovine embryos on survival after deep freezing and on sex ratio. Bovine IVF embryos and bovine oviductal epithelial cells (BOEC) were co-cultured in TCM-199 and, on Day 7 after insemination (Day 0), were morphologically evaluated and divided into groups by developmental stage. In Experiment 1, embryos classified as early blastocysts, blastocysts and full-expanding blastocysts were randomly subdivided into 2 groups by replicate: 50% of the embryos were placed immediately in a new BOEC co-culture (fresh group), while the other 50% were frozen, thawed and placed in a new BOEC co-culture (frozen/thawed group). Embryos were frozen in 1.5 M glycerol using a standard slow cooling technique. Fresh and frozen/thawed embryos were compared for survival rate (embryos hatching/hatched) in BOEC co-culture over the following 3 d (i.e., Days 7 to 10). The overall survival of the 425 embryos (early to full-expanding blastocysts) was 33% and was not different between fresh (35%) and frozen/thawed (30%) embryos. Survival of embryos cultured fresh or after freezing/thawing was higher for full-expanding blastocysts than for early blastocysts or for blastocysts, both of which were not different. In Experiment 2, all frozen/thawed embryos used in Experiment 1 plus all morulae and hatched blastocysts collected and frozen on Day 7 without regard to survival were sexed utilizing the polymerase chain reaction (PCR) technique. Sex of the embryos, by stage of development on Day 7, was determined in order to compare the rate of development in BOEC co-culture with the sex ratio (percentage of males). A total of 235 embryos was sex-determined with an overall percentage of males of 51%, which was not different from the expected 1:1 sex ratio. Both full-expanding blastocysts and hatched blastocysts had a significantly higher (P < 0.05) proportion of males (68 and 100%, respectively), while morulae had a significantly lower proportion of males (24%). Early blastocysts and blastocysts did not differ from a 1:1 sex ratio. The results indicate that male embryos develop faster in vitro than female embryos. The higher survival rate of full-expanding blastocysts after freezing/thawing, and the production of a higher number of males than females among embryos of this developmental stage suggest that a greater number of male fetuses may result from the successful freezing and transfer of in vitro-produced bovine embryos.

Journal Article↗
PubMed2001

Detection of transgene in progeny at different developmental stages following testis-mediated gene transfer.

We recently reported that exogenous DNA injected into testis as a liposome complex can be transferred into the egg via sperm by natural mating and integrated in the genome (testis-mediated gene transfer: TMGT). Here, we studied the efficiency of each of the several liposomes in associating foreign DNA with sperm, the expression of an introduced gene in early embryos, and the presence of the DNA in fetuses and pups at different ages. The CMV/beta-actin/EGFP fusion gene, encapsulated with different liposomes, was injected into rat testis, and spermatozoa in the cauda epididymis were obtained 1, 4, and 14 days after injection. We tested each of the 8 liposomes, and found that only 2, DMRIE-C and SuperFect, led to the detection of foreign DNA on all of the days examined, with relatively higher ratios of rats having positive sperm. By means of TMGT using either of those two liposomes, more than 80% of morula-stage embryos expressed EGFP, as observed by fluorescence microscopy. Then we detected introduced DNA in the progeny by PCR and Southern dot blot, and found that the ratio of animals carrying the foreign DNA decreased as they developed, and that only a part of postpartum progeny were foreign-DNA-positive with high incidence of mosaicism. These results suggest that, although, the success rate is still limited, foreign DNA could be integrated into the genome of the progeny by TMGT at least under specific experimental conditions, the efficiency of which depends largely on the characteristics of the liposome. The results also suggest that TMGT could be applicable to fetal gene therapy as well as to the generation of transgenic animals.

Animals↗
PubMed1990

[Effect of pigmented epithelium from different developmental stages of chick embryo on cell growth of retinal explants].

The model system to investigate the effect of retinal pigmented epithelium (PE) on the retinal development in vitro has been established in this laboratory. Chick retina separated from 5-day-old embryo (E 5) were cut into strips and explanted on the collagen substratum either in close contact with retinal PE (RPE), or without PE (R). The rates of cell proliferation of retinal strips cultured for 48 hr were measured by the uptake of radioactive thymidine and DNA contents. Both parameters in RPE were increased to values ranging from 137 to 167% when PE was taken from E5 and E6. However PE taken from E7, E8 and E9 had no effect on cell proliferation. The rate of cell proliferation of retina were increased both when separated retina and PE of E5 either from same or from an other eye closely contact again and when retina and PE of E5 were explanted together without separation. However the rates of cell proliferation were remained without much change when a millipore filter existed between retina and PE of E5 as well as the retina was inverted, the ganglion cell layer contacted with PE. The neurite outgrowth from retina explant with and without PE of E5 or E6 were also different. After culture for 24 hr the fiber length of neurite growing in RPE was only 36-39% of that in R. After 48 hr it was about 70% of that in R. This results suggested that the developmental stage of PE and the direct cell-to-cell contact of PE from E5 with photoreceptor layer of retina was important for the retinal cell proliferation. But PE had negative influence on neurite growth of retina in culture.

Animals↗
PubMed1998

Membrane-bound cysteine proteinase isoforms in different developmental stages of Trypanosoma cruzi.

Cysteine proteinase isoforms, immunologically cross-reactive with cruzipain and with a similar apparent molecular mass, have been identified in epimastigotes of Trypanosoma cruzi by extraction and phase partition using the detergent Triton X-114. These isoforms are concentrated in the microsomal fraction obtained after differential centrifugation, which is known to consist essentially of plasma membrane, can be labelled by incubation of live parasites with sulfo-NHS-biotin, and bind to cystatin-sepharose affinity columns. They are present, albeit with a different electrophoretic pattern, in the epimastigote, amastigote and trypomastigote stages of the parasite.

Animals↗
PubMed2003

Plasmodesmal changes are related to different developmental stages of antheridia of Chara species.

During the development of the antheridia of Chara species, dynamic changes in the occurrence and ultrastructure of plasmodesmata are observed which are closely correlated to particular developmental phases and presumably regulate the morphogenetic events in the antheridia. The disappearance of plasmodesmata between shield cells and between shied cells and the basal cell leads to a cessation in symplasmic transport around the antheridum and determines its concentric or centrifugal character via centrally situated capitular cells. Unplugged plasmodesmata are present between fully synchronously developing antheridial filament cells and obviously coordinate the development of the cells. In the middle phase of spermiogenesis, rough endoplasmic reticulum in antheridial filaments passes uncompressed through wide plasmodesmata and provides an additional transport pathway for developmental control factors. Plugged plasmodesmata link cells of different types or cells of the same type which are at different phases of cell cycle and guarantee their individual development. The plugging of plasmodesmata is a reversible process that depends on the morphogenetic situation. Plasmodesmata connecting the basal cell and the subbasal cell as well as the basal cell and capitular cells are transformed successively from the simple into the complex type and might be the pathways for an import of gibberellins and nutrients into the strong sink tissues of the developing antheridium. There is a symplasmic connection between the antheridum and the thallus via a basal cell. Prior to the initiation of spermatozoid differentiation (spermiogenesis), plasmodesmata connecting the basal cell with a subbasal cell and the basal cell with capitular cells are spontaneously broken, resulting in symplasmic isolation of the antheridium that is probably a signal which triggers the induction of spermatozoid differentiation. Premature plasmolytically evoked symplasmic isolation of the antheridium leads to the elimination of 1 to 2 cell cycles from the proliferative stage of spermatogenesis. Autoradiographic studies demonstrate that both natural and induced symplasmic isolation drastically decreases the entry of isotopically labeled gibberellic acid into antheridia of Chara species that may be the consequence of the elimination of the hormone's transport through plasmodesmata.

Autoradiography↗
PubMed2005

Gene discovery and expression profile analysis through sequencing of expressed sequence tags from different developmental stages of the chytridiomycete Blastocladiella emersonii.

Blastocladiella emersonii is an aquatic fungus of the chytridiomycete class which diverged early from the fungal lineage and is notable for the morphogenetic processes which occur during its life cycle. Its particular taxonomic position makes this fungus an interesting system to be considered when investigating phylogenetic relationships and studying the biology of lower fungi. To contribute to the understanding of the complexity of the B. emersonii genome, we present here a survey of expressed sequence tags (ESTs) from various stages of the fungal development. Nearly 20,000 cDNA clones from 10 different libraries were partially sequenced from their 5' end, yielding 16,984 high-quality ESTs. These ESTs were assembled into 4,873 putative transcripts, of which 48% presented no matches with existing sequences in public databases. As a result of Gene Ontology (GO) project annotation, 1,680 ESTs (35%) were classified into biological processes of the GO structure, with transcription and RNA processing, protein biosynthesis, and transport as prevalent processes. We also report full-length sequences, useful for construction of molecular phylogenies, and several ESTs that showed high similarity with known proteins, some of which were not previously described in fungi. Furthermore, we analyzed the expression profile (digital Northern analysis) of each transcript throughout the life cycle of the fungus using Bayesian statistics. The in silico approach was validated by Northern blot analysis with good agreement between the two methodologies.

Amino Acid Sequence↗
PubMed1996

Toxic effect of ethanolic extract of Nerium oleander (Apocynaceae) leaves against different developmental stages of Muscina stabulans (Diptera-Muscidae).

Nerium oleander (Apocynaceae) is evergreen shrubs widely used for ornamental purpose in mediterranean region. The present investigation, revealed for the first time the insecticidal effect of ethanolic extract from leaves of this plant against 2nd instar larvae of the medically important false stable fly Muscina stabulans (Diptera: Muscidae). LC50 of the extract was 113.66 ppm. This dose delayed larval and pupal duration suppressed oviposition and decreased adult longevity of the survivors. Morphogenic abnormalities were recorded and photographed in larval, pupal and adult stages, which were produced from treating 2nd instar larvae with different concentrations of the extract.

Animals↗
PubMed2002

Content and composition of phosphoglycerols and neutral lipids at different developmental stages of the eggs of the codling moth, Cydia pomonella (Lepidoptera: Tortricidae).

Phosphoglycerol, triacylglycerol, diacylglycerol, and free fatty acid content was studied in eggs of the codling moth Cydia pomonella at the white, red ring, and black head developmental stages. The composition of total phosphoglycerols and of the three classes of neutral lipids was also analyzed. The highest total lipid content was found in eggs at the white stage, the amount decreasing during development mainly as a result of a diminution in the quantity of phosphoglycerols, which account for approximately 50% of total content at all stages of egg development. The amount of triacylglycerols and free fatty acids changes significantly during development, whereas only minor changes were found in diacyglycerol levels. The total phosphoglycerol acyl composition of eggs at the white and red ring stages is similar, whereas differences are evident at the black head stage of development. Triacylglycerols and free fatty acids are enriched in saturated fatty acids in all analyzed stages. The acyl profile of diacylglycerols is different at each stage. The unsaturation index decreases in diacylglycerols and free fatty acids as a function of egg development. The results of the present paper suggest that triacylglycerols may constitute an important source of energy during the final period of egg development while phosphoglycerols may function as fuel during the beginning. Phosphoglycerols could be precursors for the triacylglycerol biosynthesis that takes place between white and red ring stages.

Animals↗
PubMed1997

Angiotensin II receptors: localization of type I and type II in rat epididymides of different developmental stages.

Previous studies from our laboratory have provided evidence for the existence of a local renin-angiotensin system in the rat epididymis. Evidence has also accumulated, indicating that locally formed angiotensin II from the rat epididymis may play a paracrine and/or autocrine role in regulating epididymal electrolyte and fluid transport. In the present study, specific anti-peptide antibodies against the second extracellular loops of angiotensin II type I (AT1) and type II (AT2) receptors were used to localize immunocytochemically these receptors in the rat cauda epididymides of three developmental stages, namely, immature (2-week), early mature (6-week) and fully mature (10-week). The immunostaining intensity for AT1 receptors was found to be stronger than that for AT2 receptors throughout rat epididymides of all stages. However, the immunostaining for both AT1 and AT2 receptors observed in the fully mature rat epididymis was much more intense than that observed in the epididymides of the two younger stages. While the immunostaining for both AT1 and AT2 receptors in the younger rat epididymides appeared to be distributed in both basal and apical regions, the immunostaining in the fully mature epididymis was predominantly localized in the basal region. The present finding of the differential patterns of angiotensin II receptor immunoreactivity in three different stages of the rat epididymis may reflect the fine tuning of rat epididymal function by angiotensin II, acting as a paracrine or autocrine agent, during the course of development.

Amino Acid Sequence↗
PubMedDate not supplied

Comparative studies of the induction of somatic eye-color mutations in an unstable strain of Drosophila melanogaster by MMS and X-rays at different developmental stages.

An unstable white locus in Drosophila melanogaster originally described by Rasmuson and Green (1974) and further by Rasmuson et al. (1978, 1980) contains an IS element. This constellation interacts with the zeste mutation and forms a mutationally unstable system that is sensitive to a variety of mutagens. Mutational shifts between zeste and wild-type eye color as well as deletions and transpositions of the white locus are frequently occurring in the unstable X-chromosome in germ line and in somatic tissue. Germinal mutations from zeste to wild-type eye color are associated with an insertion of a piece of DNA, proximal to the wsp site, and the shifts from red to zeste are caused by an excision of the same piece (Rasmuson, in preparation). Mutations to pigmentless phenotype are interpreted as deletions of the white locus, while they always are irreversible and show non-complementation with wsp. The somatic system can be used as a screening test for potential mutagens, described by Rasmuson et al. (1984). This survey is an attempt to correlate the size of the mutated area of the eyes with the age of the larvae at mutagen treatment. X-Rays and MMS were used to give an indication of the mechanism of the instability, according to the different kinds of DNA damage induced. The results show that the mean size of red spots decreased with increasing age of larvae at treatment, while the mutation frequencies were increased because of the multiplication of the cells in the eye anlage susceptible to the mutagens. This is contradictory to the hypothesis maintained by Fahmy and Fahmy (1980) that the somatic shifts are not mutagenic but epigenetic events, due to altered regulation of the gene expression. Red spots induced with MMS are smaller in size than X-ray-induced red spots, indicating a delay in the establishment of mutations from chemically-induced lesions compared to irradiation damage. White spots on the other hand were equally large in size, irrespective of inducing agent and about twice the size of the chemically-induced red spots, implying a faster and more direct action for fixation of deletions than for the production of MMS induced shifts in eye color from zeste to red.

Age Factors↗