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The dendritic translocation of translin protein in the form of BC1 RNA protein particles in developing rat hippocampal neurons in primary culture.

Neural BC1 RNA is distributed in neuronal dendrites as ribonucleoprotein particles (RNP). Our previous studies indicated the presence of Translin in BC1 RNPs, which is a translational repressor and links a subset of mRNAs to microtubules. In this study, we confirmed that Translin associates with BC1 RNP and we used immunocytochemical methods to examine the subcellular distribution of Translin in developing hippocampal cells in primary cultures. Translin was detected in both the nuclei and cytoplasm of neurons, whereas in glial cells it was localized in the nuclei. Consistent with the reported developmental time course of BC1 RNA expression and dendritic delivery the translocation of Translin to the neuronal dendrites appeared to correlate with neuronal development and differentiation events such as the onset of synaptogenesis in culture. These observations suggest that BC1 RNP or Translin itself may be relevant to the dendritic translation of mRNAs in response to transsynaptic activity.

Animals↗

Expression and function of the ultraspiracle (usp) gene during development of Drosophila melanogaster.

The usp locus encodes a member of the nuclear hormone receptor superfamily in Drosophila melanogaster that interacts with EcR (ecdysone receptor) to mediate ecdysteroid-induced gene expression. A 2.7-kb usp mRNA was detected at all developmental times tested, although its abundance varied. Among premetamorphic stages, both the 2.7-kb transcript and Usp protein attained their highest levels in the late third larval instar. The 2.7-kb usp transcript was also found in adult stages and a 1.2-kb transcript was detected in the polyadenylated RNA fraction of both mature adult females and early embryos. Aneuploids carrying two usp mutant alleles and a putative variegating usp+ allele often developed deformities of the adult wing disc that apparently resulted from mutational disruption of usp activity before metamorphosis and whose frequency was affected by maternal genotype. Both of the recessive lethal usp mutations associated with this "cleft thorax" phenotype involved substitutions of conserved arginine residues in the DNA-binding domain, although the frequency of the phenotype was not the same for the two alleles. Both mutant proteins retained the ability to form heterodimers with EcR in vitro but showed reduced affinity for an ecdysone response element.

Amino Acid Sequence↗

Collapsin-1/semaphorin D is a repellent for chick ganglion of Remak axons.

Chick collapsin-1/human semaphorin III/mouse semaphorin D is believed to guide the extension of specific axons by a repellent mechanism. Here we examine its role in the guidance of axons of the ganglion of Remak (Remak) in the developing chick intestine. Early in embryogenesis Remak axons extend parallel to, but do not enter, the intestine when collapsin-1 is expressed in the adjacent rectal wall. Remak axons later penetrate the peripheral portions of the rectal wall when collapsin-1 expression retreats from the outer muscle layer to the more internal submucosal and mucosal layers of the rectum. Extension of Remak neurites is repelled in vitro by rectum explants and also by 293T cells expressing collapsin-1. The rectal chemorepellent activity is blocked by anti-collapsin-1 antibodies. Our results suggest that collapsin-1 may help prevent Remak axons from projecting into the intestinal wall at early developmental times and later restricts Remak axon trajectories to the outer part of the intestinal muscle layer.

Alkaline Phosphatase↗

The role of ABI3 and FUS3 loci in Arabidopsis thaliana on phase transition from late embryo development to germination.

Arabidopsis abi3 and fus3 mutants are defective in late embryo development and their embryos show precocious growth. To understand the function and role of ABI3 and FUS3, we analyzed expression patterns of genes which were normally activated during late embryo development and germination in these mutants. Using the differential display method, both upregulated and downregulated genes were observed in immature siliques of the abi3 fus3 double mutant. Four clones having more abundant expression in the abi3 fus3 double mutant than in wild type were isolated. These genes were activated during wild-type germination, suggesting that some genes that are activated during wild-type germination are precociously activated in the abi3 fus3 mutant during late embryo development. Also, genes that were activated during wild-type germination were isolated and their expression patterns during late embryo development in the wild type and in abi3, fus3, and abi3 fus3 mutants were analyzed. Sixteen such clones were found, and 11 of these showed derepression or precocious activation of gene expression in the mutants. These results indicate that ABI3 and FUS3 negatively regulate a particular set of genes during late embryo development. We also showed that immature fus3 siliques accumulated one-third of the wild-type level of abscisic acid (ABA), but mature fus3 siliques accumulated ABA at a level comparable to that in the wild type. The possible mechanisms of controlling developmental timing in late embryo development as well as collaborative and distinct roles of ABI3 and FUS3 are discussed.

Arabidopsis↗

Developmental change in the steroid hormone signal for cell-autonomous, segment-specific programmed cell death of a motoneuron.

During metamorphosis of the hawkmoth, Manduca sexta, accessory planta retractor (APR) motoneurons undergo a segment-specific pattern of programmed cell death (PCD): e.g., APRs from abdominal segment six [APR(6)s] die at pupation in direct response to the prepupal rise in 20-hydroxyecdysone (20E), whereas APR(4)s survive through the pupal stage and die at eclosion (adult emergence). The hypothesis that the death of APR(4)s is triggered by the decline in 20E at eclosion was supported by findings that injection of 20E into developing pupae to delay the fall in 20E delayed APR(4) death. Furthermore, abdomen isolation to advance the fall in 20E caused precocious APR(4) death. In other experiments, APR(4)s were placed in primary cell culture 4 days before eclosion in medium containing 1 microg/ml 20E. A switch to hormone-free medium induced PCD in a significant proportion of APR(4)s, compared to APR(4)s that remained in 20E. Process fragmentation was a reliable early indicator of PCD. These results show that a decline in 20E triggers cell-autonomous PCD of APR(4)s, in contrast to the rise in 20E that triggers cell-autonomous PCD of APR(6)s. Thus, the PCD of homologous motoneurons in different body segments at different developmental times is triggered by different steroid hormone signals.

Animals↗

The origin of mesoderm in phoronids.

Descriptive studies of phoronid development have concluded that the mesoderm of these animals originates from the endoderm during gastrulation. This interpretation has been tested by labeling one blastomere of 4- through 16-cell embryos and examining the position and germ layers occupied by the labeled clones of cells in the larva. No 2 injections gave rise to identical clones of cells, suggesting that the cleavage program does not generate cells of unique identity and that cell fates are established at later developmental time points. In many cases, a relatively large sector composed of ectodermal cells was labeled. When these labeled cells were adjacent to the mouth or anus of the larva, muscle and mesenchyme cells originated from the labeled clones. Under these circumstances, nerve cells also originated from these labeled sectors. These labeling studies also showed that endodermal cells can give rise to mesodermal and neural cells. These results suggest that nerve and muscle cells are induced to form at ectodermal-endodermal boundaries from both germ layers. These marking experiments also confirmed the observation that nerve cells originate both from the apical organ and the trunk region and show for the first time that the intestine originates by ingression of posterior ectoderm.

Animals↗

Drosophila C virus: experimental study of infectious yields and underlying pathology in Drosophila melanogaster laboratory populations.

The underlying pathology of a nonhereditary virus, the Drosophila C virus, was studied. This study was related to the contamination routes (ingestion or contact) and developmental timing. When oral contamination occurred at the first larval instar: (1) the flies were contaminated, (2) the flies which had developed the most rapidly were the most infected, (3) in newly emerged females, the level of virus was higher than in newly emerged males, (4) when infected flies were reared on virus-free medium only males lost their virus. Moreover, oral contamination of adults was very efficient, but the highest virus yield was obtained when both larvae and imagos grew on virus-contaminated medium. About 30 to 50% of the flies died on the sixth day. They were as DCVC invaded as DCVC-injected flies. It seemed that when the virus yield was higher than a given threshold, all flies died, whatever had been the contamination routes. When contaminated adult females and virus-free males were reared together on a virus-free medium, females could infect males. In contrast, contaminated males were not able to infect virus-free females. Thus, only females were able to contaminate a rearing medium or other flies.

Animals↗

Impact of a North American isolate of the microsporidium Nosema carpocapsae on a laboratory population of the codling moth, Cydia pomonella.

Nosema carpocapsae is a microsporidian pathogen of the codling moth, Cydia pomonella. We report the occurrence of this pathogen in a colony originating from collections made in the United States. This is the first record of N. carpocapsae infecting North American codling moths. This North American isolate of N. carpocapsae was indistinguishable from isolates received from New Zealand and Bulgaria, based on small subunit ribosomal RNA sequencing, but was more virulent than the previously described New Zealand isolate. In the laboratory, infected larvae and pupae had increased mortality compared to their uninfected counterparts and developmental time increased by 1 week. There was no effect on female fecundity. Within a cohort of eggs laid by infected females, neonates that emerged first were more likely to be uninfected. We established an uninfected colony by interrupting horizontal transmission and only utilizing the larvae that emerged from the first-laid eggs.

Animals↗

Schwann cell CD9 expression is regulated by axons.

The cell-surface glycoprotein, CD9, has been implicated in hematopoietic and Schwann cell signaling in vitro. In vivo, Schwann cell CD9 expression follows a developmental time course that parallels that of myelin genes. Here we report that Schwann cell CD9 mRNA expression is regulated by axonal contact in culture and in vivo. Following adult rat sciatic nerve injury, CD9 mRNA expression in distal nerve sections is correlated with the presence of axons; CD9 is down-regulated when axons are degenerating and is reexpressed when axons are regenerating, but not if they are prevented from doing so by transection. CD9 regulation in cultured Schwann cells is also dependent on the presence of neurons such that dissociated Schwann cells down-regulate CD9 mRNA expression in culture, but in the presence of sensory neurons they continue to express CD9. Therefore, regulation of Schwann cell CD9 expression parallels that of myelin genes protein 0 and myelin basic protein in culture and in vivo. A signaling role for CD9 in nerve development and regeneration is proposed.

Animals↗

Bayesian Phylogenetic Lineage Reconstruction with Loss of Heterozygosity Mutations Derived from Single-Cell RNA Sequencing.

Mutations are acquired frequently, such t`hat each cell's genome inscribes its history of cell divisions. Loss of heterozygosity (LOH) accumulates throughout the genome, offering large encoding capacity for phylogenetic inference of cell lineage.In this chapter, we demonstrate a method, using single-cell RNA sequencing, for reconstructing cell lineages from inferred LOH events in a Bayesian manner, annotating the lineage with cell phenotypes, and marking developmental time points based on X-chromosome inactivation. This type of retrospective analysis could be incorporated into scRNA-seq pipelines and was initially developed to investigate Emx1+ cortical projection neuron and glia lineages from C57Bl/6J (B6) and CAST/EiJ (CA) interstrain F1 mice, describing progenitor cells giving rise to multiple cortical cell types through stereotyped expansion and distinct waves of neurogenesis.

Animals↗

Gene transfer of troponin I isoforms, mutants, and chimeras.

Thin filament proteins play an essential role in the regulation of myocardial pressure development. Within the thin filament of the sarcomere, troponin I (TnI) plays a key role in regulating the Ca(2+) sensitivity of force. During myocardial development, there is a transition in TnI isoform expression from the slow skeletal isoform (ssTnI) in embryonic/fetal myocardium to the cardiac isoform (cTnI) expressed in adult hearts. Over a similar developmental time window, the calcium sensitivity of force development also decreases. Gene transfer of ssTnI, and chimeras derived from ssTnI and cTnI, into adult ventricular myocytes have provided insights into the isoform-specific domains of TnI responsible for differentially influencing myofilament Ca(2+) sensitivity. Two separate isoform-specific regions, located in the carboxyl- and amino-portions of the protein, have been identified by comparing Ca(2+)-activated isometric tension in myocytes expressing the TnI isoforms or chimeras. The carboxyl-portion of TnI also contributes to isoform-dependent differences in myofilament sensitivity to acidic pH, which ensues during several myocardial disease states. In contrast, the diminished Ca(2+) sensitivity observed in response to beta-adrenergic-mediated phosphorylation of cardiac TnI requires the amino-portion of the cardiac TnI isoform yet, does not depend on the presence of a specific isoform in the carboxyl-region of TnI. Recent studies with a mutation linked to hypertrophic cardiomyopathy have demonstrated that changes in protein charge also influence the ability of TnI isoforms to regulate myofilament Ca(2+) sensitivity. Information gained from these, and future studies on more localized and specific changes in the amino acid sequence, may one day lead to the use of genetically engineered TnI for therapeutic manipulation of contractile function.

Actin Cytoskeleton↗

A case of abnormal size in eggs and larvae of the Taiga tick, Ixodes persulcatus (Acari: Ixodidae).

Egg batches laid by females of Ixodes persulcatus (maintained under a photoperiod of L:D 20:4 before feeding) contained eggs of two sizes: numerous normal eggs of 0.578 x 0.421 mm and some giant eggs of 0.776 x 0.515 mm. Giant eggs were 32-34% greater in length and 22-24% greater in width than normal eggs. Females maintained under a short-day photoperiod (L:D 12:12) laid only normal eggs (0.588 x 0.417 mm). There were no differences between these eggs in viability and developmental time, but larvae hatched from giant eggs were larger (length of body 23-28% greater and of gnathosoma 12-15% greater) than larvae from normal eggs. This dimorphism of eggs and larvae was not sex determined, but may be a maternal photoperiodic effect.

Animals↗

Chronic toxicity of cadmium to Chironomus riparius (Diptera: Chironomidae) at different food levels.

The interacting effects of cadmium toxicity and food limitation on the midge, Chironomus riparius, were studied during chronic exposure in laboratory experiments. If the food was supplied ad libitum, both larval developmental time and mortality of the larvae were negatively affected by cadmium concentrations of 2.0-16.2 micrograms/L. The number of eggs deposited per female and the mean life span of the imagines were not affected by cadmium. Integration of these separate effects into a population growth rate showed a clear reduction with increasing cadmium concentrations. Food limitation of unexposed larvae at high population density reduced fitness, judged on all parameters studied and consequently reduced the population growth rate (up to 85%). The effects on larvae, which were exposed to both cadmium and food limitation, differed considerably from the response to the individual stress factors. Exposure to cadmium increased mortality among food-limited first and second instar larvae. Consequently, the amount of food available for each surviving larva increased. At the two lowest concentrations studied (2.0 and 5.6 micrograms Cd/L), these indirect positive effects of cadmium overruled the direct negative effects and caused an increase of the fitness of the food-limited exposed larvae compared to the food-limited, unexposed controls. At a concentration of 16.2 micrograms Cd/L, the negative effects of cadmium on food-limited midges balanced the positive effects of reduced food limitation. At this concentration, the population growth rate did not differ significantly from the food-limited control any more. It is concluded that the indirect positive effects of cadmium on food limitation could eliminate negative, direct effects of low cadmium concentrations on food-limited chironomid populations.

Administration, Oral↗

Development of spectral mechanisms in the ground squirrel retina following lid opening.

The retina of the California ground squirrel (Spermophilus beecheyi) contains three classes of photopigments (lambda max = 440, 500, 525 nm). From optic nerve recordings it was previously discovered that the effectiveness of signals from the 440 nm cone increase gradually over the weeks following lid opening. In this experiment several features of the electroretinogram (ERG) were examined to assess developmental changes in signals originating in the outer retina. As judged by threshold sensitivity, suprathreshold responsivity, or adaptability, the contribution of the 525 nm mechanism reaches its adult level two to three weeks after lid opening. The 500 nm mechanism appears to have a similar developmental time course. The development of the 440 nm mechanism was tracked using a chromatic adaptation measure. So indexed, the time required for this mechanism to reach its adult status was much greater than that for the other two mechanisms (70-80 days vs 40-45 days of age). The relatively slow development of effectiveness of the 440 spectral mechanism appears to reflect events occurring in the outer retina.

Adaptation, Physiological↗

Effects of neonatal intraocular colchicine on synaptogenesis and on the retention of the ipsilateral retinofugal projection within the superior colliculus.

A normally transient ipsilateral retinofugal projection exists in the rat but is retained following eye removal because of the loss of competitive interaction between crossed and uncrossed fibers. To further explore this phenomenon, colchicine (10(-3)M) was injected into the right eye of newborn albino rats to partially suppress axonal transport in optic fibers, alter the developmental time course of retinofugal synaptic terminals and determine if this would in turn extend the period of survival of the ipsilateral projection. Measurements of the number of fibers in the nerve were also made to insure that colchicine was not lethal to the retinofugal projection. Projections into the superior colliculus were demonstrated by anterograde movement of HRP from the left eye. TMB histochemistry revealed dense labeling of the contralateral retino-recipient layers at 5 dpn in untreated or saline-injected controls. The ipsilateral projection was seen as a lighter band of activity across the colliculus which was most concentrated in the antero-medial quadrant. This pathway was transient and degenerated by 10 dpn, except for a few antero-medial fibers. Animals treated with colchicine demonstrated a retention of this pathway through 20 dpn. A concomitant quantitative analysis of synaptic development within the superior colliculus revealed populations of boutons with round (R) and flattened (F) vesicles, as well as multiple junctional (MJ) and serial (S) complexes, most of which were specialized R boutons. The various synaptic categories displayed specific ratios unique to the different stages of maturation. Intraocular colchicine reduced the ratio of R, MJ and S boutons to F terminals between 5-15 dpn (P less than 0.01). By 20 dpn, the proportions of MJ and S boutons remained depressed but the normal ratio of R to F boutons was restored. Areal determinations of each synaptic profile included in the counts revealed a significant reduction in the size of MJ synaptic profiles examined in colchicine-treated animals and this may have been reflected in the slight loss of tectal volume (6-9%). Removal of the left eye and assessment of degenerating boutons showed that the expanded ipsilateral projection was not sufficiently dense to produce such a restoration. It thus appears that colchicine delayed the growth of the R population, but the effect was reversible and this category of boutons continued to develop, albeit on a later time course.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Developmental expression of dystrophin on the rat myocardial cell membrane.

The Duchenne muscular dystrophy gene product, dystrophin, is expressed on the cell membrane of skeletal and cardiac muscles. We examined the developmental time course of dystrophin expression in the rat ventricular myocardium. Dystrophin was immunohistochemically undetectable on the 15th embryonic day, but a small amount was expressed on the cell membrane on the 17th embryonic day. The amount increased during the perinatal period reaching adult level at 2 weeks after birth. These findings were supported by immunoblot analysis. Chemical sympathectomy in newborn rats by 6-hydroxydopamine (prevention of innervation) had no detectable influence on myocardial dystrophin expression at 2 weeks after birth. Our results show that dystrophin lining on the rat myocardial cell membrane increases during the perinatal period, and that this process is little influenced by the development of sympathetic innervation.

Aging↗

Expression of cholinergic markers in transplants of immature mouse neocortex into adult mouse parietal cortex.

Cerebral cortex of embryonic and newborn mouse was removed and transplanted into adult mouse neocortex to investigate whether the development of cholinergic markers would proceed normally after transplantation. It is shown that both AChE and muscarinic receptors developed in transplanted mouse neocortex. However, while muscarinic receptor binding increased to adult levels in transplants from both pre- and postnatal donors AChE staining intensity only achieved adult levels in tissue from postnatal donors. These results are in accordance with the normal developmental time course of the two different cholinergic markers, since muscarinic receptors appear prenatally in cortex while AChE becomes histochemically detectable after birth. These data suggest that the prenatal donor tissue, unlike the postnatal transplants, lacks an environmental signal for the appearance of AChE stained elements, that is, presumably host AChE fiber ingrowth. Transplantation of immature donor CNS tissue into adult host thus appears to be a useful paradigm to study the regulation of differentiation processes.

Animals↗

[Electron microscope studies on development stages of Theileria annulata (Dschunkowsky, Luhs, 1904) in the intestine and haemolymph of Hyalomma anatolicum excavatum (Koch, 1844) (author's transl)].

For the first time developmental stages of Theileria within the intestine and haemolymph of ticks were studied by electron microscopy. On the 1st-6th day post repletionem the intestine of the ticks contained humerous tages, which were very similar to microgamonts and microgametes of haemosporidia. Later, on the 14th-19th day p.r. ookinete-like stages ere found in the haemolymph of ticks. From these results it seemed likely that gamogony of Theileria takes place in the intestine of ticks. In more detail: The microgamont-like stages were spindle-shaped; they measured about 8-12 mu in length with a diameter of about. 08 mu in their middle region...

Animals↗