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Cyclic AMP induces the synthesis of developmentally regulated plasma membrane proteins in Dictyostelium.

Dictyostelium discoideum slugs (pseudoplasmodia) were disaggregated and the resynthesis of developmentally regulated plasma membrane proteins examined. The synthesis of the majority of these proteins was inhibited when cells were overlaid with Cellophane and maintained as a monolayer. However, cell contact and movement did occur under the Cellophane. The inhibition of differentiation may result from the inability of the cells to organize specifically into multicellular aggregates. The addition of cyclic AMP (1--5 mM) induced the synthesis of certain developmentally regulated plasma membrane proteins in cells overlaid with Cellophane. Hence, this confirms other work showing that cyclic AMP is required for at least some post-aggregative gene expression. Specific cell organisation and interactions are apparently required for an increase in or maintenance of intracellular cyclic AMP levels.

Cell Membrane

Subunit composition and molecular weights of the developmentally regulated lectins from Dictyostelium discoideum.

Discoidin I, the developmentally regulated carbohydrate binding protein from Dictyostelium discoideum, migrated as a broad band during electrophoretic analysis using 10% polyacrylamide gels in the presence of sodium dodecyl sulfate. The band could be resolved into two components by electrophoresis on 15% polyacrylamide gels. All highly purified preparations of discoidin I contained equal amounts of the two components, suggesting they are nonidentical subunits of the protein. The subunit molecular weights of discoidin I and discoidin II were found to be 29,500 and 26,500, values somewhat larger than those previously reported.

Carrier Proteins

Effects of a post-translational modification mutation on different developmentally regulated glycosidases in Dictyostelium discoideum.

We have studied the effect of a post-translational modification mutation upon four developmentally regulated glycosidases of Dictyostelium discoideum. The presence of the modA mutation affects the intracellular level of these multimeric enzymes differently. The level of alpha-glucosidase is unaffected in the modA mutant. The mutant cell contains only a very small fraction of the wild type beta-glucosidase-1 activity. The alteration in modification renders beta-glucosidase-1 holoenzyme thermolabile and susceptible to degradation in vivo. alpha-Mannosidase-1 and N-acetylglucosaminidase are found at approximately 1/3 of the wild type level in the modA mutant. Degradation of holoenzyme does not appear to be responsible for the low level of these activities. We propose that alpha-mannosidase-1 and N-acetylglucosaminidase subunits are being degraded prior to subunit assembly. We conclude the modification bestows different properties upon the various glycosidases.

Acetylglucosaminidase

Use of common plant lectins for isolation and characterization of constitutive and developmentally regulated cell surface associated glycoproteins of Dictyostelium discoideum.

Glycoproteins as a class of molecules have been implicated as serving crucial roles in cell recognition events. Using 3 common plant lectins, we have isolated and identified a number of cell surface associated glycoproteins. The appearance of at least 5 of these proteins is under developmental regulation.

Chromatography, Affinity

Developmentally regulated lectins in cellular slime molds and embryonic chick tissues.

Several species of cellular slime mold (including D. discoideum and P. pallidum) and a number of embryonic chick tissues (including muscle, heart, brain, and liver) contain lectin activities that can be extracted and assayed as hemagglutinins. In all cases studied the lectin activities show significant changes with differentiation. The studies with cellular slime molds are more advanced; and suggest that lectins play a role in developmentally regulated cell cohesion. The function of the embryonic chick lectins in differentiation is presently under investigation.

Animals

Developmentally regulated induction of neurite outgrowth from immature chick sensory neurons (DRG) by homogenates of avian or mammalian heart, liver and brain.

Neurite outgrowth is elicited from whole explants or dissociated neurons of 8--10-day-old chick embryo sensory, dorsal root ganglia when cultured in the presence of a high speed supernatant fraction (105,000 g) from homogenates of chick or rat heart, liver or brain. The neurite promoting activity is not identical to mouse nerve growth factor (NGF) and is non-dialyzable. Expression of this neurogenic factor would appear to be developmentally regulated as its activity is barely detectable in organs from 6--9-day-old embryos but specific activity rises dramatically in homogenates of organs from embryos of greater than 11 days incubation. Greatest activity is found in chick heart and rat brain with only trace levels in lung or kidney and none in spinal cord.

Animals

In planta genome editing in citrus facilitated by co-expression of CRISPR/Cas and developmental regulators.

Recent advances in the field of genome editing offer a promising avenue for targeted trait improvements in fruit trees. However, the predominant method taken for genome editing in citrus (and other fruit trees) involves the time-consuming tissue culture approach, thereby prolonging the overall citrus breeding process and subjecting it to the drawbacks associated with somaclonal variation. In this study, we introduce an in planta approach for genome editing in soil-grown citrus plants via direct transformation of young seedlings. Our editing system, abbreviated here as IPGEC (in planta genome editing in citrus), is designed to transiently co-express three key gene groups in citrus tissue via Agrobacterium tumefaciens: (i) a genome-editing catalytic group, (ii) a shoot induction and regeneration group, and (iii) a T-DNA enhanced delivery group. This integrated system significantly improves de novo shoot induction and regeneration efficiency of edited tissue. By incorporating single-guides RNA's (sgRNA's) targeting the carotenoid biosynthetic gene PHYTOENE DESATURASE (CsPDS), the IPGEC system effectively produced mutated albino shoots, confirming its ability to generate homozygous/biallelic genome-edited plants. By using high throughput screening, we provide evidence that transgene-free genome-edited plants could be obtained following the IPGEC approach. Our findings further suggest that the efficiency of specific developmental regulators in inducing transformation and regeneration rates may be cultivar-specific and therefore needs to be optimized per cultivar. Finally, targeted breeding for specific trait improvements in already successful cultivars is likely to revolutionize fruit tree breeding and will pave the way for accelerating the development of high-quality citrus cultivars.

Citrus

Developmental regulation of progenitor aging shapes long-term intestinal homeostasis in Drosophila.

Aging causes disruption of tissue homeostasis, with stem cell exhaustion as a major hallmark. However, whether aging trajectories are established during development remains unexplored. Here, we demonstrate that genetic modulation of aging-associated pathways in larval adult midgut progenitors (AMPs) determines the trajectory of Drosophila adult intestinal homeostasis. Induction of aging-associated pathways in the AMPs results in aberrant proliferation, skewed differentiation, barrier dysfunction, and genomic instability. Ultimately, AMP islet architecture is destabilized and age-related molecular signatures are altered. In contrast, reversing aging-associated effects results in a decrease in the enteroendocrine population and the barrier is unaffected. Together, our findings demonstrate that aging-associated pathways are tightly regulated during early development and perturbation can hamper adult gut homeostasis, establishing AMPs as key developmental determinants.

Drosophila