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J Palka

Publications and source records attributed to J Palka.

17 recordsLinked to original sources

The development of normal and ectopic sensilla in the wings of hairy and Hairy wing mutants of Drosophila.

We have utilized enhancer trap markers to follow the development of ectopic sensillar precursors in the wings of Drosophila induced by the mutations hairy and Hairy wing. Normal sensilla are still present in these mutations, and can be distinguished from ectopic sensilla based upon both the position and the timing of their development. This correlates well with the development of ectopic achaete expression in these mutations: such staining is detected only after the appearance of normal staining. Thus, neither mutation appears to alter the specification of proneural clusters in the wing, as identified with anti-achaete, or the specification of sensillar precursors within these clusters. Rather, both act to induce the formation of a temporally and spatially distinct phase of sensillar development.

Animals

Guidepost cells.

Guidepost cells, as classically defined in the grasshopper embryo have only rarely been found in other systems. If the concept of guidepost cells is expanded, recognizing that any special role of specific cells in axon guidance is a function of the entire landscape in which axons are growing, and that growth cone--guidepost interactions may share mechanisms with many other cell--cell interactions, then numerous examples are found in both the peripheral and central nervous systems of many species.

Animals

A student apparatus for recording action potentials in cockroach legs.

A comparatively simple apparatus allows even beginning students to observe action potentials in the cockroach leg. The recordings are made extracellularly by impaling the leg on two insect pins. Deflection of large spines on the leg, which are each innervated by one sensory neuron, initiates the action potentials. Using this technique, students observe the all-or-nothing nature of action potentials, their coding of information by frequency, and sensory adaptation.

Action Potentials

Elevated activity of low molecular weight insulin-like growth factor-binding proteins in sera of vitamin C-deficient and fasted guinea pigs.

We have previously reported that scorbutic and fasted guinea pig sera contain an insulin-like growth factor-I (IGF-I)-reversible inhibitor of collagen, proteoglycan, and DNA synthesis in cultured cells. Here we report that IGF-binding protein (IGFBP) activity is increased in serum containing the inhibitor [125I]IGF-I or -II bound to these sera was eluted in the 30- to 50-kDa region of an S200 gel column. [125I]IGF-I affinity cross-linking analysis revealed that a 38-kDa cross-linked species increased markedly in fasted and scorbutic sera, with a lesser increase in a 34-kDa species, while scorbutic sera also yielded a 44-kDa species. Gel filtration of unlabeled sera showed a 10-fold increase in the activity of two proteins in the 30- to 50-kDa region from the experimental sera. Their activity correlated with their ability to inhibit binding of [125I]IGF-I to its cellular receptor, suggesting that they have the potential to inhibit IGF-I-dependent functions. Ligand blotting showed that 29 and 35-kDa IGFBPs were almost undetectable in normal serum, but were dramatically induced by scurvy and fasting, so that they accounted for close to 40% of the total circulating BPs. Total IGFBP-3 in the experimental sera was increased about 30%, while there was little effect of scurvy or fasting on the level of BP-3 activity isolated by acid extraction of the high mol wt region of the S200 column. An IGF-I analog with normal affinity for the 30- to 50-kDa BPs from fasted and scorbutic sera, but with reduced affinity for the cell receptor, was equivalent to IGF-I in reversing the inhibition of collagen synthesis by scorbutic guinea pig serum in human fibroblasts. Thus, reversal of inhibition appears to require initial saturation of IGFBPs. The overall results suggest that two circulating IGFBPs with unoccupied binding sites are induced in vitamin C-deficient or fasted guinea pigs and may be responsible for inhibition of IGF-I-dependent functions by sera from these animals.

Animals

Scorbutic and fasted guinea pig sera contain an insulin-like growth factor I-reversible inhibitor of proteoglycan and collagen synthesis in chick embryo chondrocytes and adult human skin fibroblasts.

Chick embryo chondrocytes cultured in sera from scorbutic and fasted guinea pigs exhibited decreases in collagen and proteoglycan production to about 30-50% of control values (I. Oyamada et al., 1988, Biochem. Biophys. Res. Commun. 152, 1490-1496). Here we show by pulse-chase labeling experiments that in the chondrocyte system, as in the cartilage of scorbutic and fasted guinea pigs, decreased incorporation of precursor into collagen was due to decreased synthesis rather than to increased degradation. There was a concomitant decrease in type II procollagen mRNA to about 32% of the control level. As in scorbutic cartilage, proteoglycan synthesis by chondrocytes in scorbutic serum was blocked at the stage of glycosaminoglycan chain initiation. Scorbutic and fasted guinea pig sera also caused a 50-60% decrease in the rates of collagen and proteoglycan synthesis in adult human skin fibroblasts, which synthesize mainly type I collagen. Decreased matrix synthesis in both cell types resulted from the presence of an inhibitor in scorbutic and fasted sera. Elevated cortisol levels in these sera were not responsible for inhibition, as determined by the addition of dexamethasone to chondrocytes cultured in normal serum. Insulin-like growth factor I (IGF-I, 300-350 ng/ml) reversed the inhibition of extracellular matrix synthesis by scorbutic and fasted guinea pig sera in both cell types and prevented the decrease in type II procollagen mRNA in chondrocytes. Therefore, in addition to its established role in proteoglycan metabolism, IGF-I also regulates the synthesis of several collagen types. An increase in the circulating inhibitor of IGF-I action thus could lead to the negative regulation of collagen and cartilage proteoglycan synthesis that occurs in ascorbate-deficient and fasted guinea pigs.

Animals

Neurogenic and antineurogenic effects from modifications at the Notch locus.

The best studied mutations at the Notch locus produce a neurogenic phenotype, with a massive overgrowth of the nervous system at the expense of epidermis. We report here that, in the development of the adult peripheral nervous system, the Abruptex alleles of Notch have the opposite phenotype, namely an underproduction of sensory organs or sensilla. This arises primarily not from an arrest of the lineages that produce sensilla, from the degeneration of sensillar cells, or from the transformation into neurons of cells that normally secrete the cuticular components of a sensillum (as can happen in Notch alleles). Rather, our evidence argues strongly that the sensillar mother cells never form. This implies that the Notch protein plays a role in the process that first generates a difference between sensillar mother cells and ordinary epidermal cells. The number of sensilla formed on the wing of flies carrying multiple doses of Notch+ is virtually the same as that of wild type, i.e. the Abruptex phenotype is not reproduced to any significant extent. This suggests that the single amino acid substitutions that occur in Abruptex mutants confer on the protein some functionally distinctive feature, possibly more powerful intermolecular binding or altered stability.

Alleles

Genes involved in the development of the peripheral nervous system of Drosophila.

The development of the peripheral nervous system (PNS) requires the activity of a number of genes. The neurogenic and the proneural genes are necessary in the earliest phase; their mutations lead to hyperplasia and partial or total elimination of the PNS respectively. Some of these mutations also affect other developmental processes. Other mutations affect later events: cut transforms one type of sensory organ into another; numb alters the fate of the components of a single sensory organ. We will describe the effects of the best studied mutations on PNS development and discuss the possible role of the wild type genes.

Animals

Mosaic Drosophila wings reveal regional heterogeneity in the guidance of ectopic axons.

In most studies of axon guidance in the peripheral tissues of insects, the ability of experimentally perturbed axons to pathfind was examined only along their normal pathways. This means that regions normally devoid of axons have not been sampled for their ability to influence axonal trajectories. To examine this question, we have induced the formation of single sensory neurons in a variety of abnormal locations in the developing wing of Drosophila and have examined the course taken by their axons. The axons of such ectopic neurons have a regionally varying tendency to grow in the normal, proximal direction. This proximal bias approaches 100% for neurons located in the distal part of vein L2 and 70% in distal vein L4 but falls to chance (50%) along vein L5. Thus, neurons forming in ectopic regions of the wing, especially those found near the normal axon pathways (veins L1 and L3), have a high probability of growing axons in the correct direction. We conclude that information relevant to axon outgrowth is not restricted to the normal pathways. Whether this information is intrinsic or extrinsic to the neurons, and why its strength shows such conspicuous regional variation, awaits further study.

Animals

Similar hormonal changes in sera from scorbutic and fasted (vitamin C-supplemented) guinea pigs, including decreased IGF-I and appearance of an IGF-I reversible mitogenic inhibitor.

We previously proposed that the decreased rates of synthesis of collagen and proteoglycans in vitamin C-deficient guinea pigs were unrelated to the role of ascorbate in proline hydroxylation but might result from modulation of hormones known to change during fasting. In the present studies, we found that sera from guinea pigs on an ascorbate-free diet for 24-28 days or from those fasted for 4 days, with vitamin C supplementation, showed similar changes in the concentrations of several hormones. EGF and IGF-II concentrations were unchanged, but cortisol was increased 3-5 times and growth hormone was increased to approximately twice normal levels. Thyroxine and IGF-I concentrations were decreased to 40% and 25-33% of normal levels, respectively. The decrease in serum IGF-I must occur by a growth hormone-independent pathway. The extent of changes in hormone concentrations in sera from ascorbate-deficient guinea pigs was correlated with the extent of weight loss. Sera from scorbutic and fasted guinea pigs failed to stimulate DNA synthesis in quiescent BALB 3T3 cells in the presence of saturating concentrations of EGF and PDGF. Addition of experimental sera to normal serum showed that lack of mitogenic activity was due to the presence of an inhibitor. Inhibition was not related to IGF-I concentrations in the sera, although it was reversed by the addition of IGF-I to sera from scorbutic or fasted animals. These results support our proposed model and suggest that IGF-I, as well as an inhibitor of its activity, plays a role in the regulation of growth by vitamin C and other nutrients.

Animals

Salt stimulation of serum insulin-like growth factor binding protein activity.

Insulin-like growth factors (IGF)-I and -II are bound to carrier or binding proteins in serum. There are at least two classes of binding protein: a high molecular weight complex and a low molecular weight species that is relatively unsaturated. Total binding capacity in serum generally is determined by incubating [125I]IGF with protein that has been stripped of IGF by acid gel filtration. We found that addition of NaCl to the assay increased binding to stripped guinea pig binding protein to about two to four times the level measured in the absence of salt. Stimulation by NaCl was optimal between concentrations of 0.6 and 1.4 M and also was observed when fetal calf or human sera were used as sources of stripped binding protein or when IGF-II was the ligand. Using chloride salts, the order of activity with respect to cations was Na+ greater than K+ greater than Li+. Na2HPO4 at 0.6 M was as stimulatory as 1.2 M NaCl but 0.6 M Na2SO4 was less effective. NH4HCO3 was as effective as NaCl at 0.6 M. Scatchard plots of data from competitive dilution experiments with [125I]IGF-I and unlabeled IGF-I showed that binding was heterogeneous in the absence of 0.6 M NaCl but linear in its presence. NaCl did not stimulate binding when whole serum was used, but after gel filtration of serum on Sephacryl 200 at pH 8, which does not dissociate IGFs from binding protein, binding to individual fractions was stimulated three- to fourfold by NaCl. Fractions stimulated included those containing the large complex or the unsaturated binding protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Axonal polarity in Drosophila wings with mutant cuticular polarity patterns.

In Drosophila melanogaster certain mutations alter the polarity of trichomes and bristles, cuticular structures secreted by the epithelial cells of the adult fly. Since sensory neurons arise from epithelial cell precursors, and sensory axons grow along the inner faces of epithelial cells, we have studied the developing wings of these mutants to see whether the change in epithelial cell polarity has an influence on the direction of axon outgrowth. The nerve patterns formed in the mutants prickled, inturned, and frizzled, however, were largely normal, indicating that in these cases the polarity of the cuticular structures produced by the epithelial cells is altered without any effect on the polarity of the associated axons.

Animals

Deafferentation slows the growth of specific dendrites of identified giant interneurons.

The effect deafferentation has on the morphology of giant interneurons was studied in the abdominal nervous system of crickets (Acheta domesticus). The morphology of four uniquely identified giant interneurons was exxamined by iontophoresing cobalt chloride into the neurons of interest. A major source of afferents for these interneurons consists of mechanoreceptors located on paired abdominal sensory appendages -- the cerci. Partial deafferentation of the giant interneurons was obtained by pinching off the cercus at hatching and maintaing the specimen in this deprived condition until adulthood. The interneurons of three groups of animals were examined; control specimens which were not treated surgically, unilaterally treated specimens which had a single cercus removed and bilaterally treated specimens which had both cerci removed. Two types of morphological changes were detected. (1) Chronic removal of a cercus was correlated with a reduction in length of dendrites ipsilateral to the ablated cercus; however, the general form of the dendritic branching pattern remained constant and recognizable. Two dendrites of a single neuron could be influenced independently if they were innervated by separate cerci. Thus deprivation did not have a generalized effect on growth of a neuron, rather it specifically influenced the dendrites deprived of afferents. (2) It was also observed that the projection of cercal sensory fibers in specimens reared with a single cercus differed from normal in that scattered fibers cross the midline in regions of the ganglion where none usually exist. It is suggested that modifications in the response properties of these deprived neurons are based on these two changes in morphology.

Animals

Central connections of receptors on rotated and exchanged cerci of crickets.

The cerci of crickets, paired abdominal appendages bearing sound-sensitive filiform hairs, can be removed and grafted back so that their morphological axes acquire various relationships to those of the body. We have studied both the morphogenetic consequences of such surgery and the central connections made by the regenerating axons of the cercal sensory neurons. If a cercus is rotated and grafted back into its own socket, it back-rotates towards its original orientation in succeeding molts. If left and right cerci are exchanged, with or without rotation, back-rotation does not occur and super-numerary cerci are formed in predictable locations. There are two sub-populations of filiform hairs: those that vibrate transversely to the cercal shaft (T-hairs) in dorsal and ventral sectors, and longitudinally vibrating hairs (L-hairs) in lateral and medial sectors. Two giant interneurons are excited by T-hairs of their own side but not by L-hairs. If cerci are grafted so that they assume various orientations relative to the body, a consistent physiological result is obtained: T-hairs always appear to be the source of excitatory input to the giant interneurons, no matter where they are caused to be located by prior surgery. The phenomena of back-rotation, formation of supernumerary cerci, and formation of connections selectively by T-hairs, can be interpreted on the hypothesis of morphogenetic gradients.

Animals