Pump In Response Research Articles

Indirect Flow Rate Estimation of the NEDO PI Gyro Pump for Chronic BVAD Experiments

In totally implantable ventricular assist device systems, measuring flow rate of the pump is necessary to ensure proper operation of the pump in response to the recipient's condition or pump malfunction. To avoid problems associated with the use of flow probes, several methods for estimating flow rate of a rotary blood pump used as a ventricular assist device have been studied. In the present study, we have performed a chronic animal experiment with two NEDO PI gyro pumps as the biventricular assist device for 63 days to evaluate our estimation method by comparing the estimated flow rate with the measured one every 2 days. Up to 15 days after identification of the parameters, our estimations were accurate. Errors increased during postoperation days 20 to 30. Meanwhile, their correlation coefficient r was higher than 0.9 in all the acquired data, and estimated flow rate could simulate the profile of the measured one.

IRI-1, a LIN-15B homologue, interacts with inositol-1,4,5-triphosphate receptors and regulates gonadogenesis, defecation, and pharyngeal pumping in Caenorhabditis elegans.

Inositol-1,4,5-triphosphate receptors (IP(3)Rs) are ligand-gated Ca(2+) channels that control Ca(2+) release from intracellular stores. They are central to a wide range of cellular responses. IP(3)Rs in Caenorhabditis elegans are encoded by a single gene, itr-1, and are widely expressed. Signaling through IP(3) and IP(3)Rs is important in ovulation, control of the defecation cycle, modulation of pharyngeal pumping rate, and embryogenesis. To further elucidate the molecular basis of the diversity of IP(3)R function, we used a yeast two-hybrid screen to search for proteins that interact with ITR-1. We identified an interaction between ITR-1 and IRI-1, a previously uncharacterized protein with homology to LIN-15B. Iri-1 is widely expressed, and its expression overlaps significantly with that of itr-1. In agreement with this observation, iri-1 functions in known itr-1-mediated processes, namely, upregulation of pharyngeal pumping in response to food and control of the defecation cycle. Knockdown of iri-1 in an itr-1 loss-of-function mutant potentiates some of these effects and sheds light on the signaling pathways that control pharyngeal pumping rate. Knockdown of iri-1 expression also results in a sterile, evl phenotype, as a consequence of failures in early Z1/Z4 lineage divisions, such that gonadogenesis is severely disrupted.

Bioenergetics of the formyl-methanofuran dehydrogenase and heterodisulfide reductase reactions in Methanothermobacter thermautotrophicus.

The synthesis of formyl-methanofuran and the reduction of the heterodisulfide (CoM-S-S-CoB) of coenzyme M (HS-CoM) and coenzyme B (HS-CoB) are two crucial, H2-dependent reactions in the energy metabolism of methanogenic archaea. The bioenergetics of the reactions in vivo were studied in chemostat cultures and in cell suspensions of Methanothermobacter thermautotrophicus metabolizing at defined dissolved hydrogen partial pressures ( pH2). Formyl-methanofuran synthesis is an endergonic reaction (DeltaG degrees ' = +16 kJ.mol-1). By analyzing the concentration ratios between formyl-methanofuran and methanofuran in the cells, free energy changes under experimental conditions (DeltaG') were found to range between +10 and +35 kJ.mol-1 depending on the pH2 applied. The comparison with the sodium motive force indicated that the reaction should be driven by the import of a variable number of two to four sodium ions. Heterodisulfide reduction (DeltaG degrees ' = -40 kJ.mol-1) was associated with free energy changes as high as -55 to -80 kJ.mol-1. The values were determined by analyzing the concentrations of CoM-S-S-CoB, HS-CoM and HS-CoB in methane-forming cells operating under a variety of hydrogen partial pressures. Free energy changes were in equilibrium with the proton motive force to the extent that three to four protons could be translocated out of the cells per reaction. Remarkably, an apparent proton translocation stoichiometry of three held for cells that had been grown at pH2<0.12 bar, whilst the number was four for cells grown above that concentration. The shift occurred within a narrow pH2 span around 0.12 bar. The findings suggest that the methanogens regulate the bioenergetic machinery involved in CoM-S-S-CoB reduction and proton pumping in response to the environmental hydrogen concentrations.

A Direct Interaction between IP3 Receptors and Myosin II Regulates IP3 Signaling in C. elegans

Molecular and physiological studies of cells implicate interactions between the cytoskeleton and the intracellular calcium signalling machinery as an important mechanism for the regulation of calcium signalling [1–11]. However, little is known about the functions of such mechanisms in animals. A key component of the calcium signalling network is the intracellular release of calcium in response to the production of the second messenger inositol 1,4,5-trisphosphate (IP3), mediated by the IP3 receptor (IP3R) [12–14]. We show that C. elegans IP3Rs, encoded by the gene itr-1, interact directly with myosin II. The interactions between two myosin proteins, UNC-54 and MYO-1, and ITR-1 were identified in a yeast two-hybrid screen and subsequently confirmed in vivo and in vitro. We defined the interaction sites on both the IP3R and MYO-1. To test the effect of disrupting the interaction in vivo we overexpressed interacting fragments of both proteins in C. elegans. This decreased the animal's ability to upregulate pharyngeal pumping in response to food. This is a known IP3-mediated process [15]. Other IP3-mediated processes, e.g., defecation [16], were unaffected. Thus it appears that interactions between IP3Rs and myosin are required for maintaining the specificity of IP3 signalling in C. elegans and probably more generally.

Regulated disruption of inositol 1,4,5-trisphosphate signaling in Caenorhabditis elegans reveals new functions in feeding and embryogenesis.

Inositol 1,4,5-trisphosphate (IP(3)) is an important second messenger in animal cells and is central to a wide range of cellular responses. The major intracellular activity of IP(3) is to regulate release of Ca(2+) from intracellular stores through IP(3) receptors (IP(3)Rs). We describe a system for the transient disruption of IP(3) signaling in the model organism Caenorhabditis elegans. The IP(3) binding domain of the C. elegans IP(3)R, ITR-1, was expressed from heat shock-induced promoters in live animals. This results in a dominant-negative effect caused by the overexpressed IP(3) binding domain acting as an IP(3) "sponge." Disruption of IP(3) signaling resulted in disrupted defecation, a phenotype predicted by previous genetic studies. This approach also identified two new IP(3)-mediated processes. First, the up-regulation of pharyngeal pumping in response to food is dependent on IP(3) signaling. RNA-mediated interference studies and analysis of itr-1 mutants show that this process is also IP(3)R dependent. Second, the tissue-specific expression of the dominant-negative construct enabled us to circumvent the sterility associated with loss of IP(3) signaling through the IP(3)R and thus determine that IP(3)-mediated signaling is required for multiple steps in embryogenesis, including cytokinesis and gastrulation.

Effect of short-term cyclic stretch on sodium pump activity in aortic smooth muscle cells.

We previously demonstrated that expression of both the alpha1- and alpha2-subunits of Na+-K+-ATPase is elevated after a 2- to 4-day cyclic stretch in aortic smooth muscle cells. In this study, we determined the effect of short-term (2-30 min) cyclic stretch on the activity of the Na pump and investigated possible mechanisms that may be involved in the action of stretch. Na pump activity was significantly increased above the baseline activity between 2 and 30 min of stretch. This effect of stretch was reversible within 1 h. Intracellular Na was also elevated at corresponding time points. Blocking the entry of Na with Gd and amiloride did not affect the stretch-induced increase in Na pump activity. Inhibition of protein kinase A (PKA) activity attenuated the effect of stretch on the Na pump. Furthermore, inhibition of polymerization of actin and phosphatidylinositol 3-kinase (PI3K) activity prevented the action of stretch on Na pump activity. We conclude that the stimulation of the Na pump in response to cyclic stretch requires the integrity of the actin cytoskeleton as well as the activity of PI3K, which has a role in intracellular vesicular trafficking. PKA may also be involved in this effect of stretch on Na pump.

Adaptive regulation of the canalicular bile salt export pump in response to changes of bile salt pool size

Adaptive regulation of the canalicular bile salt export pump in response to changes of bile salt pool size

Altered phosphorylation of sarcoplasmic reticulum contributes to the diminished contractile response to isoproterenol in hypertrophied rat hearts.

We tested the hypothesis that changes in phosphorylation of the sarcoplasmic reticulum (SR) protein, phospholamban (PIB) and myofibrillar proteins troponin I (TnI) and C protein are responsible for the decreased relaxant response to isoproterenol in cardiac hypertrophy and failure induced by ascending aortic banding in rats. In isolated perfused heart preparations under maximal isoproterenol stimulation, the capacity for in vitro cAMP-dependent phosphorylation of PIB was significantly increased at the compensatory stage of hypertrophy (126-130%, P<0.001), but decreased with failure (70-76%, P<0.001). Phosphorylation of TnI also decreased in the failing hearts, however to a lesser extent (80-83%, P<0.05). No significant hypertrophy-related difference was evident in isoproterenol-induced phosphorylation of C protein. The relative tissue level of PIB was increased (150-168%, P<0.001) in hypertrophied and decreased (71-83.8%, P<0.05) in failing hearts compared with the respective age-matched sham-operated controls (100%). As a percentage above baseline, the maximal isoproterenol-induced increase in the EC50 of the SR Ca2+ pump in response to phosphorylation of PIB was 38.5+/-1.1% for sham-operated rats, and 26.0+/-3.8% and 15.4+/-4.2% for hypertrophied and failing hearts respectively. As a consequence, linear correlation was observed between the maximal increase in EC50 and the maximal rate of relaxation [(-dP/dt)/DevP] upon isoproterenol stimulation, declining with progressive hypertrophy to failure. These data suggest that hypertrophy-induced alterations in PIB phosphorylation and protein level contribute to the diminished relaxant response of the hypertrophied and failing heart to adrenergic agonists.

Functional expression of the Na/K pump is controlled via a cyclosporin A-sensitive signalling pathway in activated human lymphocytes

An immunosuppressant cyclosporin A (CsA) inhibits T-cell proliferation by blocking the nuclear factor of activated T-cells (NFAT) required for expression of the interleukin-2 (IL-2) gene. This work has demonstrated for the first time that in human blood lymphocytes (HBLs) activated by phytohemagglutinin (PHA), CsA at anti-proliferative doses inhibits the late sustained increase in ouabain-sensitive Rb(K) influxes, which accompanies the growth phase of G 0/G 1/S transition. CsA affects neither the initial, transient activation of the pump in response to PHA nor the ouabain-resistant ion fluxes during cell cycle progression. When the HBLs were rendered competent to proliferate by phorbol 12,13-dibutyrate ester and ionomycin in the presence of CsA, the exogenous IL-2 did not bypass the initial inhibitory effect of CsA on the long-term pump enhancement. When applied after the competence induction, CsA produced no effect on the sustained increase in ouabain-sensitive Rb influxes during the IL-2-induced progression phase. These results indicate that in activated HBLs, (1) IL-2 is involved in functional expression of the Na/K pump during cell transition from quiescence to proliferation, (2) the cell cycle-associated upregulation of the pump is related to a CsA-sensitive signalling pathway.

Metabolism of trout red blood cells: correlation between cation transport and oxygen uptake following adrenergic stimulation

Red blood cells from rainbow trout rely mainly on oxidative phosphorylation to cover the energy demand of processes such as cation transport and protein synthesis. Stimulation of these cells with catecholamines leads to an increase in the intracellular Na + concentration due to adrenergic activation of the erythrocyte Na +–H + exchanger. Adrenergic stimulation also increases red cell oxidative phosphorylation which largely can be ascribed to increased activity of the Na +–K + pump in response to the increase in intracellular Na + concentration. Based on experiments with the Na +-ionophore, monensin, it has been suggested that adrenergic stimulation is a prerequisite for the increase in oxidative ATP-production observed when intracellular Na + is increased. Using washed red blood cells from rainbow trout, the present study examines the role for adrenergic stimulation in the metabolic response when intracellular Na + is elevated. It is shown that while monensin (5×10 −8–5×10 −6 M) causes a persistent up-to-12-fold increase in the intracellular Na + concentration, it only transiently elevates the activity of the Na +–K + pump. Furthermore, monensin (5×10 −6 M) decreased rather than increased red cell oxygen uptake. In contrast, adrenergic stimulation, which produced a 4-fold elevation in intracellular Na +, led to a 2.5-fold increase in red cell oxygen uptake and an 11-fold increase in Na +–K + pump activity. Forty percent of the increase in oxygen uptake was blocked by inhibition of the Na +–K + pump. In monensin-treated cells, adrenergic stimulation was, however, without effect on oxygen uptake and Na +–K + pump activity. These data do not support the theory that adrenergic stimulation has a direct effect on the ATP-producing processes of trout red blood cells but rather suggest that the effect of catecholamines on oxygen uptake is restricted to effects on the energy-consuming processes of the cells.

Control of the artificial heart.

The artificial heart (AH) is devoid of physiologic connections to the recipient's native feedback control loops. Control of an AH can be either passive or dynamic. Passive intrinsic control provides limited AH response to physiologic demands. Dynamic control requires the sensing of metabolic and hemodynamic signals and their incorporation into self-adjusting AH function. A single metabolic or hemodynamic parameter cannot provide sufficient data accurately to adjust AH pumping in response to varying blood flow demands. A combination of input control signals is required for reliable and flexible AH function. The selection of appropriate input control parameters and their incorporation into AH controller designs remains a critical step in the achievement of a permanent, totally implantable AH.

Properties of Proton Pumping in Response to Blue Light and Fusicoccin in Guard Cell Protoplasts Isolated from Adaxial Epidermis of Vicia Leaves.

Guard cell protoplasts (GCPs) were isolated from the adaxial epidermis of Vicia leaves. The properties of isolated adaxial GCPs (ad GCPs) were compared with those of abaxial GCPs (ab GCPs) with respect to H+-pumping activity. A saturating pulse of blue light (200 [mu]mol m-2 s-1, 30 s) induced H+ pumping in both ad GCPs and ab GCPs under red light. The maximum rate of blue-light-dependent H+ pumping was slightly higher in ad GCPs than in ab GCPs, but the magnitude of H+ pumping in ad GCPs was 68% of that in ab GCPs. H+ pumping was responsive to the second pulse, and the rate and magnitude of the pumping increased with the time between two pulses. The periods required to achieve 50% of the maximum rate were 12 and 22 min for ad GCPs and ab GCPs, respectively. The rates of blue-light-dependent H+ pumping were saturable, with half-saturation at 630 [mu]mol m-2 (21 [mu]mol m-2 s-1, 30 s) for ad GCPs and 105 [mu]mol m-2 (3.5 [mu]mol m-2 s-1, 30s) for ab GCPs. In contrast, fusicoccin, an activator of the plasma membrane H+- ATPase, induced H+ pumping with a slightly higher rate in ad GCPs than in ab GCPs. Both types of protoplast swelled similarly in response to fusicoccin. These results suggest that ad GCPs have almost the same activity for H+ pumping as ab GCPs, whereas ad GCPs require a larger number of photons to activate the H+ pump than ab GCPs.

Chemoreception in Crocodilians: Anatomy, Natural History, and Empirical Results

Anatomical and behavioral aspects of chemoreception in crocodilians are reviewed. Extant crocodilians possess valvular nostrils regulated by smooth muscles; paired nasal cavities, each with three conchae; and flaps closing the rear of the buccopharyngeal passage to channel inspired air through the nasopharyngeal duct. Olfaction is enhanced by gular pumping, whereby the floor of the pharynx is alternately lowered and raised to pulse air through the nasal cavity. Taste buds occur on the tongue, palate, and pharyngeal walls. Crocodilians in nature or in semi-natural enclosures locate distant carrion or concealed meat, implying chemoattraction to food. The American alligator exhibits underwater head-waving and mouth-opening in response to aqueous and chloroform extracts of meat. Alligators increase gular pumping in response to the scents of various meats, indicating that they detect food chemicals by olfaction. Paired gular and paracloacal glands on the integument of crocodilians are thought to produce pheromones. Juvenile crocodilians exhibit heightened gular pumping in response to airborne skin gland secretions. Observations of adults suggest that gular and paracloacal glands produce chemical signals used in mating and/or nesting activities, but behavioral responses to skin gland exudates are too poorly documented to ascribe pheromonal properties.

Solitary calcium spike dependent on calmodulin and plasma membrane Ca 2+ pump

Resealed human red cell ghosts were loaded with Fura-2, ATP, Mg 2+, and either calmodulin (CaM) or, to prevent CaM activation of the Ca 2+ pump, a synthetic peptide that antagonized endogenous CaM (an analogue of the CaM binding domain of protein kinase II, referred to as ‘antiCaM’). The ghosts reduced the cytosolic concentration of ionized calcium ([Ca 2+] i) to 193 ± 60 nM (SD, n = 15) in a medium containing 1 mM Ca 2+ and to 30 ± 27 nM (SD, n = 62) in a medium without Ca 2+ addition. Without ATP, i.e. no fuelling of the Ca 2+ pump, the [Ca 2+] i remained high (approx. 5 μM or higher). The simultaneous addition of the ionophore A23187 and Ca 2+ rapidly increased the Ca 2+ influx, which in the CaM loaded ghosts caused a solitary spike of [Ca 2+] i, reaching maximum around 2 μM within 24 ± 6 s (SD, n = 40). On the contrary, in the ghosts loaded with antiCaM, the addition of A23187 with Ca 2+ raised [Ca 2+] i during the first 2 min to a high level (2–4 μM) with no preceding spike. Pre-incubation of CaM-ghosts with Ca 2+ diminished the height of the Ca 2+ spike, and treatment with trypsin even removed the Ca 2+ spike. The trypsin treatment activated the Ca 2+ pump prior to the rise of [Ca 2+] i, making the time-consuming CaM activation unnecessary. In conclusion, the Ca 2+ spiking is dependent on a delayed CaM activation of the plasma membrane Ca 2+ pump in response to a rapid increase of Ca 2+ influx.

Comparison of a Steady Flow Pump to a Preload Responsive Pulsatile Pump in Left Atrial–to–Aorta Bypass in Canines

A unique preload responsive pulsatile pump was compared to a centrifugal pump in total cardiac support in 25-kg canines (n = 6, each group) in the left atrial-to-aorta mode during 5 h of ventricular fibrillation. With steady flow, there was immediate drop in output from 2.1 +/- 1.0 L/min to 1.4 +/- 0.3 L/min, followed by further reduction to 0.9 +/- 0.2 L/min during 5 h of ventricular fibrillation. With a pulsatile pump, there was no significant reduction from control of 2.4 +/- 0.6 L/min and no decline during 5 h of ventricular fibrillation. With steady flow, systemic vascular resistance (SVR) rose significantly from 1,762 dyne-s-cm-5 immediately on pump to 3,013 dyne-s-cm-5 at 5 h. With physiologic pulsatile flow, significant elevation of SVR did not occur. When stressed, due to diminished left atrial return, the centrifugal pump displayed line chatter and streaks of microbubbles, whereas the pulsatile pump did not. Crystalloid volume replacement with the centrifugal pump was 6.5 +/- 1.9 L, and with the preload responsive pulsatile pump, 5.6 +/- 1.3 L. It is concluded that in the left atrial-to-aorta mode during 5 h of ventricular fibrillation and with comparable volume replacement, total cardiac support of canines is associated with lower SVR with physiologic pulsatile flow and is not accompanied by line chatter and cavitation with this preload responsive pump.

Lymphatic pumping in response to changes in transmural pressure is modulated by erythrolysate/hemoglobin.

Red blood cells and lysate products (erythrolysate) are observed consistently in lymph draining acute and chronic inflammatory reactions and from tissues subjected to trauma or surgical procedures. Using hemoglobin as a marker for erythrolysate, we have measured hemoglobin in lymph up to the 10(-6) M range in a number of pathophysiological states. Data demonstrate that erythrolysate alters the pumping characteristics of lymphatic vessels. To test the effects of erythrolysate on lymphatic pumping, bovine lymphatics were suspended in an organ bath preparation with the vessels cannulated at both inflow and outflow ends. By raising the heights of the Krebs reservoir and the outflow catheters appropriately, a transmural pressure that stimulated pumping activity could be applied to the vessels. With a fixed transmural pressure of 6 cm H2O applied to the ducts, sheep erythrolysate depressed pumping activity between 40% and 100%, with dilutions containing between 10(-8) and 10(-5) M hemoglobin. Although the active principle in the red blood cells has not been characterized, evidence from precipitation purification experiments suggests that hemoglobin is an important component. Once suppressed, pumping could be restored in many but not all vessels (often to control levels) by elevating the distending pressure above 6 cm H2O. The relation between transmural pressure and fluid pumping is expressed as a bell-shaped curve, with pumping increasing up to a peak pressure (usually 8 cm H2O) and declining at pressures above this level. By comparing pressure/flow curves, we were able to ascertain that hemoglobin shifted the lymphatic function curve to the right and, on average, reduced the maximum pumping capability of the vessels. We speculate that the presence of erythrolysate/hemoglobin in lymph may modulate the ability of lymphatic vessels to drain liquid and protein from the tissue spaces.

Pharyngeal pumping continues after laser killing of the pharyngeal nervous system of C. elegans

Using a laser microbeam to kill specific subsets of the pharyngeal nervous system of C. elegans, we found that feeding was accomplished by two separately controlled muscle motions, isthmus peristalsis and pumping. The single neuron M4 was necessary and sufficient for isthmus peristalsis. The MC neurons were necessary for normal stimulation of pumping in response to food, but pumping continued and was functional in MC- worms. The remaining 12 neuron types were also unnecessary for functional pumping. No operation we did, including destruction of the entire pharyngeal nervous system, abolished pumping altogether. When we killed all pharyngeal neurons except M4, the worms were viable and fertile, although retarded and starved. Since feeding is one of the few known essential actions controlled by the nervous system, we suggest that most of the C. elegans nervous system is dispensable in hermaphrodites under laboratory conditions. This may explain the ease with which nervous system mutants are isolated and handled in C. elegans.

Suppression of fluid pumping in isolated bovine mesenteric lymphatics by Interleukin-1: Interaction with prostaglandin E 2

In addition to local physiological forces, the modulation of lymphatic pumping by chemical mediators may play an important role in the regulation of extravascular water in inflammation and shock. Since Interleukin-1 (IL-1) appears to be of major importance in the host's response to infection by mediating many inflammatory events, we thought it important to determine if this cytokine could affect the lymphatic circulation and in particular to ask whether IL-1 was capable of altering lymphatic pumping in response to changes in transmural pressure. Bovine lymphatic segments (6 to 8 cm in length) were cannulated at both ends and suspended in an organ bath preparation. The vessels were provided with Krebs solution from a reservoir. With no net driving pressure, a transmural pressure applied to the ducts elicited contractile activity and fluid pumping with increases in pumping up to 8 cm H 2O and reductions in flow above this level of distension. Human recombinant IL-1α (10 −7 to 10 −9 M) administered into the lumina of the vessels depressed pumping activity approximately 5–30% at transmural pressures between 2 and 16 cm H 2O. With limited supplies, we could only assess the effects of human recombinant IL-1β at 10 −8 M. However, it was more potent than IL-1α, inhibiting pumping at all transmural pressures with maximum suppression in the range of 70% at peak flows. The ability of IL-1 to induce prostaglandin synthesis may be one of its most important biologic functions. It is likely therefore that IL-1 and PGE 2 are closely linked and are probably present together in inflammatory lesions. With this in mind, we investigated the effects of PGE 2 alone and in combination with IL-1α. PGE 2 by itself reduced pumping at concentrations between 10 −6 and 10 −9 M. When mixed with IL-1α (both agents at a final concentration of 10 −9 M), the mixture had a marked inhibitory effect on flow, reducing pumping 50 to 70% at peak flows. The effect of the mixture of IL-1α and PGE 2 compared with the two agents administered separately was greater than the predicted additive effect at transmural pressures above 6 cm H 2O. At lower transmural pressures, however, the level of real inhibition was less than the predicted additive effect. Measurements of outflow pressures from the isolated vessels revealed only subtle changes with IL-1α or PGE 2 alone, but the administration of the two agents together produced very irregular pressure patterns illustrating a marked effect on contractile activity. We conclude that IL-1 alone or in combination with PGE 2 reduces the sensitivity of the vessels to pressure changes and decreases the maximum pumping capabilities of the vessel. We speculate that this may have an impact on the ability of the lymphatics to remove extravasated fluid and protein from the tissue spaces in inflammatory and shock states.

An improved drug infusion pump for injecting nanoliter volumes subcortically in awake rats

A new drug infusion pump capable of injecting nanoliter volumes of drug solution into the brains of awake rats has been constructed which incorporates a new “turntable” commutator and a compact tubing compressor mechanism requiring no gears. Injector cannulas inserted into guide cannulas permanently mounted in the rat's skull are connected to the drug pump by spring-protected, PE 10 tubing. Drug solutions are delivered when the pump rollers compress the drug-filled PE 10 tubing. An additional animal-activated switch and motorized mechanism rotates the drug pump in response to the animal's movements so that the PE 10 drug reservoir is not twisted. Testing the drug pump's performance in vivo with injections of 14C-nicotine into the caudate nucleus shows that drug delivery is both reliable and reproducible.

Nerve growth factor stimulates phospholipid methylation in target ganglionic neurons independently of the cyclic AMP and sodium pump responses.

Suspensions of neurons prepared from embryonic day 12 (E12) chick sympathetic ganglia were incubated with [methyl-3H]methionine in the absence of nerve growth factor (NGF). Presentation of the factor for different periods of time resulted in an approximate three-fold stimulation of radioactivity incorporated into total phospholipid, followed by a rapid decline thereafter. Both the magnitude and the time of the response were dependent on the NGF concentration used. Also examined were possible relationships of phospholipid methylation to two other short-latency responses to NGF, i.e., control of the Na+,K+-pump and elevation of cyclic AMP content. Incubation of E12 sympathetic neurons with known transmethylase inhibitors (shown to be active in the present system) failed to prevent reactivation of the Na+,K+-pump in response to NGF administration. E16 sympathetic neurons and E15 sensory neurons, which do not depend on exogenous NGF for control of their Na+,K+-pump, still show a stimulation of phospholipid methylation when challenged with the factor. Blockage of the pump with ouabain also fails to prevent a methylation response. Thus, the pump and methylation responses to NGF occur independently of each other. Intact E8 chick dorsal root ganglia, but not E12 sympathetic ganglia, display a rapid and transient rise in their cyclic AMP content when presented with NGF. At a concentration of 10 biological units/ml, NGF elicits a peak of phospholipid methylation at 4 min, and a peak of cyclic AMP at 10 min. Methylation inhibitors prevent the methylation response, but not that of cyclic AMP.(ABSTRACT TRUNCATED AT 250 WORDS)