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At least 487 records · Page 27Linked to original sources
PubMed1993

Contributions of cooling and warming rate and developmental stage to the survival of Drosophila embryos cooled to -205 degrees C.

Because of their high susceptibility to chilling injury, permeabilized Drosophila embryos can not be cryobiologically preserved by slow freezing at rates low enough to prevent the formation of intraembryonic ice. Calculations indicated that to outrun the chilling injury they must be cooled and warmed rapidly at an estimated 20,000 degrees C/min or faster. Ordinarily, such cooling rates would inevitably produce lethal intracellular ice. To prevent this, embryos must contain and be surrounded by sufficiently high concentrations of glass-promoting solutes to induce vitrification on cooling and prevent devitrification on warming. Like Steponkus et al. (Nature 345, 170, 1990) we have used ethylene glycol as the solute and have exposed permeabilized 12-h embryos to it in two steps. (Permeabilization was effected by exposing dechorionated embryos to a mixture of 0.3% 1-butanol in n-heptane for 90 or 110 s.) The two steps were (i) a 30-min exposure to 2 M ethylene glycol at 23 degrees C and (ii) a 5-min exposure to 8.5 M ethylene glycol [+/- 10% polyvinylpyrrolidone (PVP)] at 5 degrees C. The volumetric response to the first step indicates that full permeation of the 2 M glycol has been approached by 30 min. The point of the second step is to raise the intraembryonic concentration of ethylene glycol to near 8.5 M ethylene glycol by osmotic dehydration. Survival based on hatching is some 45% at this point. When 12-h embryos in 8.5 M glycol containing 10% PVP are then cooled to -205 degrees C at approximately 100,000 degrees C/min and warmed at about that rate, an average of about 12% survive (hatch), although in about half the runs 15-29% survive. Survivals in the absence of PVP are usually poorer but have been as high as 40%. Currently, 5% of the surviving larvae develop to adult flies (Steponkus et al. reported 18% hatching and 3% development to adult). Embryos that develop but do not hatch show readily detectable abnormalities in mouth parts and dorsal closure. Very high warming rates are much more critical to survival than are very high cooling rates; for example, none survive when warming is 2000 degrees C/min. The deleterious effect of slow warming is exerted between -80 and -40 degrees C. The lack of reciprocity between the effects of time spent cooling and time spent warming argues against ascribing death to chilling injury. Rather, it and other data argue for ascribing death to the devitrification during warming of cytoplasm that vitrified during cooling.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Cell-type- and developmental-stage-specific metabolism and storage of retinoids by embryonic chick retinal cells in culture.

Biological functions of retinoids in the vertebrate retina include the role of 11-cis retinaldehyde as visual pigment chromophore, and possible effects of retinoic acid in histogenesis and cell survival. Qualitative and quantitative regulation of retinoid availability for these complex processes could involve several cell types, including retinal pigment epithelium, Müller glia and retinal photoreceptors and non-photoreceptor neurons; their relative contributions, however, have not been fully elucidated. Using purified cultures, we have carried out a study of cell-type-specific metabolism and storage of retinoids in chick embryo retinal photoreceptors and other neuronal cells, as compared to those of retinal glia. Retinal glia were found to synthesize both retinoic acid and retinyl esters, and to hydrolyse the latter; they also displayed retinol dehydrogenase activities. Cultured neurons and photoreceptors also synthesized and hydrolysed retinyl esters; their capacity for retinaldehyde synthesis from a retinol or retinyl ester substrate suggested the presence of retinol dehydrogenase activity. Retinoic acid was not synthesized in differentiated neuronal cultures, although some synthesis was detectable at early culture stages when the cells were still morphologically undifferentiated. These findings indicate that cell-type-specific metabolic activities are expressed during retinal cell differentiation in vitro, and that embryonic retinal photoreceptors and nonphotoreceptor neurons are active participants in the metabolism and storage of retinoids.

Animals↗
PubMed2001

Coupling of cAMP/PKA and MAPK signaling in neuronal cells is dependent on developmental stage.

Neurite formation, an essential feature of neuronal development, is believed to involve participation of the ras-mitogen-activated protein kinase (MAPK) and cAMP-dependent protein kinase A (cAMP/PKA)-mediated signaling pathways. These pathways have been studied extensively in the rat pheochromocytoma cell line PC12, and current hypotheses suggest a single effector mechanism resulting from the convergence of cAMP/PKA and MAPK signaling. However, based on observations using a central neuronal progenitor cell line, AS583-8, there also exists evidence that the two signaling pathways may act independently resulting in neurites with differing dynamic features. In the present study, the upstream components of cAMP/PKA signaling were examined in AS583-8 cells as well as possible interactions with the MAPK pathway. We found that activation of PKA is both necessary and sufficient for the elaboration of rapidly forming processes, typical of the cAMP response. In addition, blockade of the MAPK pathway has no effect on the cAMP response, suggesting that activation of the cAMP/PKA pathway can stimulate neurite formation independent of the MAPK pathway. In order to evaluate which cell line model, PC12 vs AS583-8, best reflects the signaling features of developing central neurons, we examined interactions between cAMP/PKA and MAPK signaling in primary neuronal cultures from several brain regions. In immature cultures (1-day-old), at a point where the initiation of neurite formation is maximal, no interaction was observed. In more mature cultures (7 days old), where synaptic contacts have been established, we found a weak but reproducible activation of MAPK following stimulation of the cAMP/PKA pathway. These results suggest that cAMP/PKA and MAPK signaling act independently at the initiation of neuritogenesis but become coupled during later stages of neuronal development. Therefore, the interaction of the two pathways may be cell stage (younger vs older) specific and may participate in cellular functions that take place after initial neurite formation.

Animals↗
PubMed1992

Expression and organization of muscle specific proteins during the early developmental stages of the rabbit heart.

The expression and intracellular distribution patterns of muscle-specific proteins were studied during rabbit embryo development (7-13 dpc) using monoclonal antibodies against titin, myosin, tropomyosin and actin, as well as the intermediate filament proteins desmin, keratin and vimentin. From our panel, titin appeared to be the first muscle-specific protein to be exclusively expressed in the embryonic rabbit heart. Upon differentiation (myocyte and myotube formation), titin reorganizes from dot-like aggregates into a cross-striated pattern (in 9- to 30-somite embryos) via a transiently filamentous distribution. When the expression and organization of the other muscle proteins was studied in relation to titin, it became apparent that tropomyosin followed upon titin with respect to its exclusive expression in the heart anlagen and its organization into a striated pattern. Myosin and desmin were organized into cross-striated patterns after titin and tropomyosin, but this arrangement had not reached its final form in 13-dpc embryos. Actin, keratin and vimentin were distributed in cytoplasmic filaments in the embryonic stages we investigated. Since the first pulsations are already detected in 3-somite embryos, we conclude that the organization of titin, tropomyosin, myosin and desmin into a striated pattern does not seem to be essential for the initiation of muscle cell contraction in the heart anlagen. Furthermore, this study shows that, in comparison with studies on mouse, chick and rat, the sequence of expression of muscle-specific and intermediate filament proteins during cardiomyogenesis is species-dependent, and that their expression and organization varies in time in different regions of the developing heart.

Animals↗
PubMed1992

Developmental staging in a marsupial Dasyurus hallucatus.

In a marsupial, Dasyurus hallucatus, pouch-young of various ages from newborn to 55 days were embedded in wax and serially sectioned. On the basis of the relative development of external and internal characteristics, they were placed in the Carnegie staging system developed by Streeter and elaborated by O'Rahilly and associates. Birth occurred at stage 15, and the end of embryogenesis (stage 23) was reached about day 33. Whereas stage 23 is characterised in all eutherians by the closure of the secondary palate, this occurs before stage 15 in D. hallucatus. Since most other characters of the newborn are at a stage 15 level of development, there has been a relative acceleration of development of the secondary palate (and forelimb) in D. hallucatus that allows it to suckle and breathe at the same time. Between D. hallucatus and eutherians, there is general agreement in the sequence of development and in the relative degree of most structures at each stage. Further marsupials should be examined to see if the minor differences noted are peculiar to D. hallucatus or apply to marsupials generally.

Animals↗
PubMed1981

Developmental stages and localization of peroxidatic activity in the leucocytes of three teleost species (Cyprinus carpio L.; Tinca tinca L.; Salmo gairdneri Richardson).

The ultrastructural localization of peroxidase (PO) in the leucocytes of three teleosts (Cyprinus carpio L., Tinca tinca L., Salmo gairdneri R.) has been investigated using the 3,3'-diaminobenzidine method. In the heterophilic granulocytes the granules show a species specific structure and are PO-positive at pH 7.6. They can be traced back to small granules arising near the Golgi apparatus (GA) in the promyelocyte. They coalesce to form larger granules and gradually change into the mature type. Myelocytes contain small unreactive granules, and these represent a second granule population. Eosinophils contain one PO-positive granule type (at pH 9), and these granules show a varying density during cell maturation. Basophils are present only in the Cyprinid species, and contain unreactive granules originating from precursors displaying a weakly positive reaction at pH 7.6. The active secretory organelles (RER, GA) are PO-negative, except for a weekly positive reaction in the flocculent matrix of the inner G-cisternae. In promonocytes and monocytes the granules are unreactive, but in the macrophages PO-positive staining occurs in a few small to medium sized granules, and in large vacuoles. A least some of these latter are apparently derived from phagolysosomes containing digested erythrocytes. Thrombocytes and lymphocytes are unreactive. The successive development of PO-positive and negative granule populations in the heterophils, and the PO-reactivity of eosinophils and basophils, show some similarities to the corresponding cells in higher vertebrates, but an analogous PO-positive ("azurophil") granule type in monocytes seems to be absent.

Animals↗
PubMed1992

Comparative sensitivity of gametes and early developmental stages of a sea urchin species (Echinometra mathaei) and a bivalve species (Isognomon californicum) during metal exposures.

Bioassays were developed using sperm of a sea urchin (Echinometra mathaei), and sperm, embryos, and larvae of a bivalve species (Isognomon californicum). Sea urchin spawning was restricted to only a few months of the year and viability of sperm throughout the year varied from 5 to 75%. Sea urchin fertilization assays were affected by temporal variation in sperm viability. Spawning in this bivalve species occurs year-round and there is little temporal variation in sperm viability. Since stringent sperm:egg ratios are not required for the bivalve embryo and larval assays, these were less affected by variation in gamete quality. The relative sensitivity of the various assays were compared during exposure to three different metal pollutants: cadmium, copper, and tributyltin. Gametes and embryos were relatively resistant to cadmium toxicity, but larvae were very sensitive. With copper and tributyltin, sea urchin and bivalve fertilization assays were the least sensitive; and bivalve growth assays were the most sensitive, followed closely by the bivalve embryo assays. On the basis of sensitivity, ease and time required to conduct the assay, and salinity tolerance, the bivalve embryo assay was recommended as the overall single most reliable toxicity bioassay. However a multispecies, multidimensional approach using sperm fertilization assays as well as embryo assays should be employed, perhaps in a hierarchal manner. Larval growth assays were deemed too tedious and time consuming to be used routinely, but due to their high sensitivity, should still be considered as a valuable comparative tool.

Animals↗
PubMed1992

Long-term effects of heavy metals in food on developmental stages of Aiolopus thalassinus (Saltatoria: Acrididae).

Newly hatched F1 nymphs of Aiolopus thalassinus (Fabr.) were fed on food treated with various concentrations of HgCl2, CdCl2, and PbCl2 until the end of adult life. Toxicological observations were followed in the F1 generation and in the F2 generation derived from the heavy metal-loaded F1 parents. The highest concentration of the heavy metal caused 100% mortality of the F1 adults within four weeks. The nymphal duration of the F1 and F2 generations was significantly prolonged after Hg and Cd exposure, but the F1 of the group treated with lead was not affected. The fresh body weight of adults was significantly reduced in the F1 generation of most treatments and in the resulting untreated F2. The lifespan of the F1 adults was shortened. In the F2 generation, although the lifespan was somewhat longer, generally it was still shorter than that of the control adults. The mean egg number laid by F1 adults fed on food contaminated with Hg or Cd was decreased. This decrease was more pronounced in the case of Cd than Hg. In the females fed on food treated with Pb the reduction of the number of egg pods was not remarkable. The hatchability of the eggs laid by F1 females was significantly reduced as compared to the control. The viability of the eggs laid by F2 adults was somewhat decreased due to either a reduced number of egg pods or to a lower rate of hatchability, especially in the case of Cd. The treated adults frequently displayed weakness in their legs, difficulties in walking, tremors, and nervous movements.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Effect of initial developmental stage on morphology of transplanted embryonic chick pancreas.

It has been reported that only certain types of pancreatic parenchymal cells survive transplantation. This study examines whether the extent of differentiation of the pancreas at the time of transplantation affects the resulting morphology or viability of its components. Segments of chick pancreas or its primordia from stages preceding formation of dorsal bud (60 h) through hatching (day 21) were implanted in the abdominal region of three-day chick embryos. After various periods of growth, grafts were examined by light- and electron microscopy. In all transplants, individual endocrine cells (A, B, D, PP) and islet structure were identical to those of normal embryos of comparable age. The exocrine portion also appeared normal in implants from embryos younger than seven days. In grafts from older donors, however, normal acini and ducts were replaced by dilated, irregularly shaped tubules from which new islets appeared to develop. These results suggest that transplantation causes structural modification of exocrine tissue, which may reflect its initial functional capabilities. These observations are compared with similar structural alterations that occur following experimental obstruction of ducts and in human pancreatic pathology.

Animals↗
PubMed1990

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↗
PubMed1976

Histone content in relation to amount of heterochromatin and developmental stage in three species of Drosophila.

Relative amounts of various histone fractions in Drosophila chromatin were estimated densitometrically on electrophoretic gel separations. Several consistent and highly significant differences were obtained between larval and adult chromatin. The arginine-rich histones showed the most conspicuous changes: higher amounts of H4 in larvae, higher H3 in adults. The level of modification of these histones was clearly higher in larval than in adult chromatin. The modification of the two slower subfractions of H4 involved, in all probability, phosphorylation as well as acetylation. In all types of Drosophila chromatin studied 50% or more of the H2a molecules were phosphorylated--a remarkably high proportion. The species differences observed in relative amounts of histone were consistent in both stages of development. D. melanogaster differed from D. hydei and D. virilis in all histones except H2b, while the latter two species were generally similar. The interspecific variation in histone pattern was generally not correlated to differences in content of heterochromatin. The level of modification of H2 was, however, presumably an exception, as it was significantly lower for both larvae and adults in D. virilis than in the other two species. These differ from D. virilis in containing appreciably lower proportions of heterochromatic chromosome segments.

Animals↗