Search PubMed⌕ Search

Biomedical subjects

C F Wu

Publications and source records attributed to C F Wu.

At least 73 records · Page 4Linked to original sources
PubMed1994

Improved stability of Drosophila larval neuromuscular preparations in haemolymph-like physiological solutions.

Neuromuscular preparations from third instar larvae of Drosophila are not well-maintained in commonly used physiological solutions: vacuoles form in the muscle fibers, and membrane potential declines. These problems may result from the Na:K ratio and total divalent cation content of these physiological solutions being quite different from those of haemolymph. Accordingly haemolymph-like solutions, based upon ion measurements of major cations, were developed and tested. Haemolymph-like solutions maintained the membrane potential at a relatively constant level, and prolonged the physiological life of the preparations. Synaptic transmission was well-maintained in haemolymph-like solutions, but the excitatory synaptic potentials had a slower time course and summated more effectively with repetitive stimulation, than in standard Drosophila solutions. Voltage-clamp experiments suggest that these effects are linked to more pronounced activation of muscle fiber membrane conductances in standard solutions, rather than to differences in passive muscle membrane properties or changes in postsynaptic receptor channel kinetics. Calcium dependence of transmitter release was steep in both standard and haemolymph-like solutions, but higher external calcium concentrations were required for a given level of release in haemolymph-like solutions. Thus, haemolymph-like solutions allow for prolonged, stable recording of synaptic transmission.

Animals↗
PubMed1994

Concomitant alterations of physiological and developmental plasticity in Drosophila CaM kinase II-inhibited synapses.

Ca2+/calmodulin-dependent protein kinase II (CaM kinase) has been implicated in neural plasticity that underlies learning and memory processes. Transformed strains of Drosophila, ala1 and ala2, expressing a specific inhibitor of CaM kinase are known to be impaired in an associative conditioning behavioral paradigm. We found that these transformants had altered short-term plasticity in synaptic transmission along with abnormal nerve terminal sprouting and directionality of outgrowth. These results represent an interesting parallel with the activity-dependent regulation of synaptic physiology and morphology by the cAMP cascade in Aplysia and Drosophila. In contrast to the learning mutants dunce and rutabaga, which are defective in the cAMP cascade, inhibition of CaM kinase in ala transformants caused increased sprouting at larval neuromuscular junctions near the nerve entry point, rather than altering the higher order branch segments. In addition, synaptic facilitation and potentiation were altered in a manner different from that observed in the cAMP mutants. Furthermore, synaptic currents in ala transformants were characterized by greater variability, suggesting an important role of CaM kinase in the stability of transmission.

Animals↗
PubMed1993

K252a potentiates epidermal growth factor-induced differentiation of PC12 cells.

Epidermal growth factor (EGF) induced short neurites in two different strains of PC12 cells. The length of the EGF-induced neurites was markedly increased in the presence of the protein kinase inhibitor K252a, which is known to inhibit differentiation induced by nerve growth factor (NGF). EGF-induced differentiation of PC12 required RNA synthesis and activity of the ras proto-oncogene product. EGF increased the levels of three neurofilament proteins and the mRNA level of two late response genes (SCG10 and 63) known to be induced by NGF. Together, EGF and K252a caused a greater increase in these mRNAs than did either agent alone. K252a did not alter the extent of EGF-induced autophosphorylation of the EGF receptor, but it did decrease the extent of receptor phosphorylation in the absence of added EGF. Thus, the ability of the EGF receptor to trigger neuronal differentiation may depend on the state of its phosphorylation at serine and/or threonine residues. Two other strains of PC12 did not extend neurites when exposed to EGF, even when K252a was also present. Thus, the differentiating effect of EGF on PC12 is PC12 strain-specific.

Animals↗
PubMed1993

Development and maintenance of a simple reflex circuit in small-patch mosaics of Drosophila: effects of altered neuronal function and developmental arrest.

A combined genetic, anatomical, and behavioral approach has been undertaken to study the developmental and functional plasticity of identified bristle mechanosensory neurons in Drosophila. A stereotyped grooming reflex in decapitated flies enabled simple but reliable assessments of the functional output of individual bristle sensory cells to correlate with their axonal projections and terminal arbors revealed by the cobalt backfill technique. Construction of small-patch mosaics that contain only a single mutant bristle allowed functional perturbation of individual neurons within an otherwise normal environment. Mutations that affect nerve excitability and membrane recycling have been used to examine their effects on neuronal pathfinding, arborization, and the initiation and maintenance of functional connections. Previous studies (Burg and Wu, 1986, J. Neurosci. 6:2968-2976; 1989, Dev. Biol. 131:505-514) have demonstrated that para(ts)nap(ts) double-mutant sensory neurons, in which action potentials are unconditionally blocked by defects in sodium currents, and eag Sh double-mutant sensory cells, in which membrane excitability is increased by alterations in potassium currents, can establish and maintain central projections that are indistinguishable from their functionally normal counterparts. Mutations of the shi(ts) gene cause a temperature-sensitive, reversible block of the membrane recycling process, resulting in arrest of neuronal growth in culture (Kim and Wu, 1987, J. Neurosci. 7:3245-3255) and depletion of synaptic vesicles that leads to transmission blockade at established synapses (Ikeda, Ozawa, and Hagiwara, 1976, Nature 259:489-491; Koenig and Ikeda, 1983, J. Neurobiol. 14:411-419; 1989, J. Neurosci. 9:3844-3860). Prolonged heat treatments (up to 16% of total development time) of small-patch shi(ts) mosaics at different pupal stages did not prevent the establishment of central projections characteristic of the various sensory cell types. However, none of the shi(ts) sensory neurons heat-pulsed during the initial or the final 16% of pupal development were able to initiate the reflex behavior, although a proportion of those treated in other periods apparently established functional contacts with appropriate targets to support the characteristic cleaning reflex. The possibility exists that the membrane recycling process blocked in shi(ts) cells provides a crucial mechanism for cell-cell interactions taking place during initial differentiation and final synaptic stabilization, and possibly competition, in the developing sensory neuron. Heat treatments of adult shi(ts) mosaics blocked the reflex initiated by the mutant (but not the surrounding normal) bristles, as expected from the effect of synaptic block.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1993

Differential ultrastructure of synaptic terminals on ventral longitudinal abdominal muscles in Drosophila larvae.

The innervation of ventral longitudinal abdominal muscles (muscles 6, 7, 12, and 13) of third-instar Drosophila larvae was investigated with Nomarski, confocal, and electron microscopy to define the ultrastructural features of synapse-bearing terminals. As shown by previous workers, muscles 6 and 7 receive in most abdominal segments "Type I" endings, which are restricted in distribution and possess relatively prominent periodic terminal enlargements ("boutons"); whereas muscles 12 and 13 have in addition "Type II" terminals, which are more widely distributed and have smaller "boutons". Serial sectioning of the Type I innervation of muscles 6 and 7 showed that two axons with distinctive endings contribute to it. One axon (termed Axon 1) has somewhat larger boutons, containing numerous synapses and presynaptic dense bodies (putative active zones for transmitter release). This axon also has more numerous intraterminal mitochondria, and a profuse subsynaptic reticulum around or under the synaptic boutons. The second axon (Axon 2) provides somewhat smaller boutons, with fewer synapses and dense bodies per bouton, fewer intraterminal mitochondria, and less-developed subsynaptic reticulum. Both axons contain clear synaptic vesicles, with occasional large dense vesicles. Approximately 800 synapses are provided by Axon 1 to muscles 6 and 7, and approximately 250 synapses are provided by Axon 2. In muscles 12 and 13, endings with predominantly clear synaptic vesicles, generally similar to the Type I endings of muscles 6 and 7, were found, along with another type of ending containing predominantly dense-cored vesicles, with small clusters of clear synaptic vesicles. This second type of ending was found most frequently in muscle 12, and probably corresponds to a subset of the "Type II" endings seen in the light microscope. Type I endings are thought to generate the 'fast' and 'slow' junctional potentials seen in electrophysiological recordings, whereas the physiological actions of Type II endings are presently not known.

Abdomen↗
PubMed1993

Ionic channels in cultured Drosophila neurons.

Cultured "giant" Drosophila neurons derived from cytokinesis-arrested embryonic neuroblasts express various membrane channels and excitability patterns. Both current- and voltage-clamp recordings could be performed on the same neuron owing to the large cell size, thus making it possible to elucidate the functional role of individual types of channels. This culture system has been used to analyze the mutational perturbations in ion channels and the resultant alterations in membrane excitability.

Action Potentials↗
PubMed1993

Removal of nitrogen and phosphorus from swine wastewater by intermittent aeration processes.

Nitrogen and phosphorus removal in a methane fermentation plus activated sludge method type pig farm (M-III) with an intermittent aeration process (IAP) was evaluated in comparison with a continuous aeration process (CAP) based on the full-scale and bench-scale experiments. Operation conditions for the treatment system were the same except for the aeration program (in the CAP), a consecutive 24-hr aeration was used, whereas in the IAP, the aeration and non-aeration periods were alternated at intervals of 3:1 hr. BOD and TOC removal efficiencies with the intermittent aeration were as high as those with the continuous operation (92-98%). In the removal of nitrogen and phosphorus, large differences between IAP and CAP were observed. At an influent T-N/TOC ratio of 0.1, removal efficiencies for T-N in the bench-scale IAP was 70%, and for T-P was 22%, respectively. At an even higher influent T-N/TOC ratios of 0.3-1.0, the removal efficiencies for T-N were decreased to about 59-61%, whereas that for T-P were -0.3-13%. In a full-scale plant, removal efficiencies for T-N with IAP and CAP were 42% and -0.09%, respectively. The results of this study show the successful performance of a simple IAP for piggery wastewater: simultaneous, one-sludge denitrification with nitrification in single-activated sludge reactor in a pig farm.

Aerobiosis↗
PubMed1993

Differential modulation of potassium currents by cAMP and its long-term and short-term effects: dunce and rutabaga mutants of Drosophila.

The cAMP concentration in Drosophila is increased by mutations of the dunce (dnc) gene and decreased by mutations of the rutabaga (rut) gene. Such mutants provide a unique means for exploring the role of cAMP in functional and developmental regulation of membrane currents. Four distinct K+ currents have been identified in Drosophila larval muscle fibers, i.e. the voltage-activated transient IA and delayed IK and the Ca(2+)-activated fast ICF and slow ICS. Results from our voltage-clamp studies indicated that both IA and IK were increased in dnc alleles. Normal muscle fibers treated with dibutyryl-cAMP showed a similar increase of IA, but no significant effect on IK. In contrast to the dnc alleles, the rut mutations appeared to enhance ICS greatly while leaving the amplitude of other currents largely unchanged. In addition, the dibutyryl-cAMP-induced increase in IA was not observed in rut fibers. Caffeine and W7, which are known to interfere with several second messenger pathways, also modulated K+ currents in larval muscle fibers. The currents in dnc and rut fibers showed strikingly altered responses to caffeine and W7. The results demonstrate that the various K+ currents in Drosophila muscles are affected by altered cAMP cascades in the mutants. The fact that not all dnc and rut mutant defects can be mimicked or reversed by acute application of cAMP suggests that long-term modulation of K+ currents by cAMP may involve mechanisms distinct from the short-term effect of cAMP.

Animals↗
PubMed1993

Cardiovascular autonomic dysfunction in non-insulin-dependent diabetes mellitus and its relation to renal dysfunction.

To investigate the relationship between cardiovascular autonomic neuropathy and nephropathy in patients with non-insulin-dependent diabetes mellitus (NIDDM), 45 patients underwent noninvasive cardiovascular reflex tests, including the Valsalva ratio, the 30:15 ratio and postural changes in systolic blood pressure (delta SBP), along with measurement of creatinine clearance (CCr) and daily protein excretion (DPE). Clinical symptoms in 30 patients were also noted and correlated with the results of the autonomic function tests. Thirty-four normal subjects underwent the same cardiovascular reflex tests and served as controls. The results showed that NIDDM patients had a significantly lower 30:15 ratio than normal subjects. However, no significant difference in either the Valsalva ratio or delta SBP was found between diabetic patients and controls. A positive correlation between the 30:15 ratio and CCr, but not DPE, was noted in diabetics. Although abnormal cardiovascular reflex tests appeared in patients who had no autonomic symptoms, abnormal test results were not parallel with the severity of symptoms. These results show that NIDDM patients have poorer cardiovascular autonomic function which may precede the appearance of autonomic symptoms. The 30:15 ratio was weakly correlated with CCr and this suggests that the incidence of cardiovascular dysfunction increases as the renal functional reserve decreases.

Adult↗
PubMed1993

Modulation of different K+ currents in Drosophila: a hypothetical role for the Eag subunit in multimeric K+ channels.

We examined the role of the ether a go-go (eag) gene in modulation of K+ currents and the possibility of its protein product Eag as a subunit in the heteromultimeric assembly of K+ channels by voltage-clamp analysis of larval muscle membrane currents. Previous DNA sequence studies indicate that the eag gene codes for a polypeptide homologous to, but distinct from, the Shaker (Sh) K+ channel subunits (Warmke et al., 1991), and electrophysiological recordings revealed allele-specific effects of eag on four identified K+ currents in Drosophila larval muscles (Zhong and Wu, 1991). Further studies of eag alleles indicated that none of the eag mutations, including alleles producing truncated mRNA messages, eliminate any of the four K+ currents, and that the mutational effects exhibit strong temperature dependence. We found that both W7, an antagonist of Ca2+/calmodulin, and cGMP analogs modulated K+ currents and that their actions were altered or even abolished by eag mutations. These results suggest a role of eag in modulation of K+ currents that may subserve integration of signals at a converging site of the two independent modulatory pathways. The Sh locus is known to encode certain subunits of the IA channel in larval muscle. The existence of multiple eag and Sh alleles enabled an independent test of the idea of Eag as a K+ channel subunit by studying IA in different double-mutant combinations. An array of allele-specific interaction between eag and Sh was observed, which reflects a close association between the Sh and eag subunits within the IA channel. Taken together, our data strengthen the possibility that the eag locus provides a subunit common to different K+ channels. The role of the eag subunit for modulating channels, as opposed to that of Sh subunits required for gating, selectivity, and conductance of the channel, suggest a combinatorial genetic framework for generating diversified K+ channels.

Animals↗
PubMed1992

Interactions of membrane excitability mutations affecting potassium and sodium currents in the flight and giant fiber escape systems of Drosophila.

We have studied the influence of the K(+)-current mutations eag and Sh and the Na(+)-current mutation napts upon two well-defined neural circuits that underlie flight and an escape response in Drosophila, recording from dorsal longitudinal and tergotrochanteral muscles. Mutations of Sh and eag affected refractory period and following frequency, but not latency, of the jump-and-flight escape response. The napts mutation altered these 3 physiological parameters of the "jump" (TTM), but not the "flight" (DLM), branch, suggesting differences in the vulnerability of different circuit components to the mutation. In contrast to their interaction in some other systems, napts did not counteract the effects of eag and Sh upon these physiological parameters in eag Sh; nap triple mutants. In eag Sh double mutants, in which multiple K+ currents may be diminished, flight muscles showed abnormal rhythmic activity not associated with flight, and some flies also had an abnormal wings-down posture. The low-frequency spikes probably originated in the flight muscle motoneurons, but the coordination between muscle fibers during this "non-flight activity" was distinct from flight. Nevertheless, in spite of the presence of this non-flight activity in resting eag Sh flies, those animals with normal wing posture were also able to fly, with a normal pattern of muscle activity. This suggests that in these mutants, the DLM motoneuron circuit is able to switch between two patterns of output, non-flight activity and flight.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1992

Synaptic plasticity in Drosophila memory and hyperexcitable mutants: role of cAMP cascade.

Activity-dependent synaptic plasticity has been implicated in the refinement and modification of neural circuits during development and learning. Previous studies show that activity-induced facilitation and potentiation are disrupted at larval neuromuscular junctions in the memory mutants dunce (dnc) and rutabaga (rut) of Drosophila. The diminished learning-memory capacity and synaptic transmission plasticity have been associated with altered cAMP levels since dnc affects the cAMP-specific phosphodiesterase and rut affects adenylate cyclase. In this study, the morphology of larval motor axon terminals was examined by anti-HRP immunohistochemistry. It was found that the numbers of terminal varicosities and branches were increased in dnc mutants, which have elevated cAMP concentrations. Such increase was suppressed in dnc rut double mutants by rut mutations, which reduce cAMP synthesis. More profuse projections of larval motor axons have also been reported in double-mutant combinations of ether à go-go (eag) and Shaker (Sh) alleles, which display greatly enhanced nerve activity as a result of reduction in different K+ currents. Therefore, we examined combinations of dnc and rut with eag and Sh mutations to explore the possible relation between activity- and cAMP-induced morphological changes. We found that the expanded projections in dnc were further enhanced in double mutants of dnc with either eag or Sh, an effect that could again be suppressed by rut. The results provide evidence for altered plasticity of synaptic morphology in memory mutants dnc and rut and suggest a role of cAMP cascade in mediating activity-dependent synaptic plasticity.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1991

Alteration of four identified K+ currents in Drosophila muscle by mutations in eag.

Voltage-clamp analysis of Drosophila larval muscle revealed that ether à go-go (eag) mutations affected all identified potassium currents, including those specifically eliminated by mutations in the Shaker or slowpoke gene. Together with DNA sequence analysis, the results suggest that the eag locus encodes a subunit common to different potassium channels. Thus, combinatorial assembly of polypeptides from different genes may contribute to potassium channel diversity.

4-Aminopyridine↗
PubMed1991

Altered synaptic plasticity in Drosophila memory mutants with a defective cyclic AMP cascade.

Synaptic transmission was examined in Drosophila mutants deficient in memory function. These mutants, dunce and rutabaga, are defective in different steps of the cyclic adenosine 3',5'-monophosphate (cAMP) cascade. In both dunce and rutabaga larvae, voltage-clamp analysis of neuromuscular transmission revealed impaired synaptic facilitation and post-tetanic potentiation as well as abnormal responses to direct application of dibutyryl cAMP. In addition, the calcium dependence of transmitter release was shifted in dunce. The results suggest that the cAMP cascade plays a role in synaptic facilitation and potentiation and indicate that synaptic plasticity is altered in Drosophila memory mutants.

Action Potentials↗
PubMed1991

Distinctions in growth cone morphology and motility between monopolar and multipolar neurons in Drosophila CNS cultures.

Growth cones play a central role in determining neurite extension, pathfinding and branching, and in establishing synaptic connections. This paper describes an initial characterization of growth cone morphology and behavior in dissociated larval central nervous system (CNS) cultures of Drosophila. Contrast-enhanced video images of growth cones in monopolar and multipolar neurons were characterized by employing morphometric parameters such as the number and length of filopodia, and the area and roundness of the lamellipodia. Behavior of growth cones was analyzed by a motility index and boundary flow plots originally devised for measuring motility in other cellular systems. We found that separate CNS regions yielded cultures of different major cell types with distinct neuritic patterns that could be correlated with the morphology and motility of the associated growth cones. Monopolar neurons were the major cell type in brain cultures, whereas multipolar neurons were predominant in ventral ganglion cultures. Moreover, the growth cones of monopolar neurons, which are likely to be associated with the axonal processes, differed from those of multipolar neurons, which might be related to dendritic terminals. Growth cones in monopolar neurons had larger lamellipodia of less erratic shape accompanied by fewer and shorter filopodia, and, when active, displayed much higher motility and less directionality in motion. Alternatively, these morphological and behavioral distinctions between monopolar and multipolar neurons may result from intrinsic differences in membrane adhesion and intracellular transport properties.

Animals↗
PubMed1991

Dual muscarinic and nicotinic action on a motor program in Drosophila.

The effect of cholinergic agonists and antagonists on the central pattern generator of the pharyngeal muscles has been studied in third instar larvae of Drosophila. The pharyngeal muscles are a group of rhythmically active fibers involved in feeding. Bath application of the cholinergic agonists carbachol, muscarine, pilocarpine, and acetylcholine (ACh) to a semiintact preparation including the pharyngeal muscles and the central nervous system (CNS), initiated long-lasting endogenous-like bursting activity in the muscles. The muscarinic antagonists, atropine and scopolamine, blocked these responses as well as endogenous activity. Perfusion with nicotine elicited a short, tonic response that was marginally blocked by mecamylamine but not by curare, alpha-bungarotoxin, hexamethonium, or the muscarinic antagonists. This is the first time that a response to cholinergic drugs has been examined in Drosophila. The pharyngeal muscle preparation may prove to be a valuable system for studying mutations of cholinergic metabolism, receptors, and second messengers.

Acetylcholine↗