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Results for “Microinjection”

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At least 451 records · Page 25Linked to original sources

PubMed1990

Action of general and alpha-smooth muscle-specific actin antibody microinjection on stress fibers of cultured smooth muscle cells.

Arterial smooth muscle cells express alpha- and gamma-smooth muscle, as well as beta- and gamma-cytoplasmic actins. Two actin antibodies, one recognizing smooth muscle and cytoplasmic actin isoforms, the other recognizing specifically alpha-smooth muscle actin, were microinjected into cultured aortic smooth muscle cells. The effect of these antibodies on stress fiber organization was examined by staining with rhodamine-labeled phalloidin and by immunofluorescence with the same antibodies. Microinjection of the general actin antibody abolished most of the stress fiber staining with all reagents, but did not significantly affect the shape of the injected cells. This suggests that stress fiber integrity is not absolutely necessary for the maintenance of cell shape within the time of observation. Microinjection of the specific alpha-smooth muscle antibody abolished to various extents the staining of stress fibers with this antibody, but left practically intact their staining with rhodamine-labeled phalloidin and with the general actin antibody. This suggests that the incorporation of alpha-smooth muscle actin is not absolutely necessary for the maintenance of stress fiber integrity in cultured smooth muscle cells.

Actins↗

PubMed1984

Microinjection of procaine or GABA into the nucleus basalis magnocellularis affects cue-elicited unit responses in the rat frontal cortex.

Male rats were chronically implanted for recording of single units in the frontal cortex during a cue-event paradigm. The rats were sedated and restrained during the experiments. Units were selected which had large-amplitude, clearly isolated action potentials. The animals were first trained to associated a 2-s tone cue with rewarding medial forebrain bundle stimulation. After training, units responded to the cue by an increase or decrease in discharge rate. Cumulative histograms of the unit response to the cue were obtained and then either procaine hydrochloride or GABA was microinjected into the nucleus basalis magnocellularis (nBM). Immediately after drug administration another histogram was obtained to ascertain the drug effect. Procaine microinjections to the nBM suppressed the frontal cortex unit responses in 9 of 10 units that had previously responded with an increase in firing rate and 10 of 12 units that had decreased their firing rate before drug administration. GABA microinjections antagonized the response in 15 of 19 excited units and 2 of 2 inhibited units. Recovery was obtained in 23 units. Other units did not remain isolated long enough to obtain complete recovery. The nBM supplies the frontal cortex with as much as 70% of its cholinergic innervation. Lesions of the region do not significantly alter the amounts of neurotransmitters other than acetylcholine in the frontal cortex. These results indicate that neurons in the nucleus basalis magnocellularis are involved in the cue-elicited changes in the rate of discharge of units in the rat frontal cortex.

Action Potentials↗

PubMed1989

Effects of unilateral microinjections of GABAergic drugs into the inferior colliculus on auditory evoked potentials and on audiogenic seizure susceptibility.

Alteration of GABAergic neurotransmission within the inferior colliculus (IC) appears to be involved in the generation of the audiogenic seizure (AGS) susceptibility. In the present study, we provide evidence indicating that this susceptibility may result from IC neuronal hyperexcitability to sound induced by a decreased GABAergic inhibition. In a first experiment, a unilateral microinjection of bicuculline, a GABAa antagonist, into the IC of normal rats increased the amplitude of the collicular auditory evoked potential, while the microinjection of THIP, a GABAa agonist, decreased this response. In a second experiment, a unilateral microinjection of bicuculline into the IC induced AGS susceptibility in normal rats.

Acoustic Stimulation↗

PubMed1986

Antiviral activity of a chemically stabilized 2-5A analog upon microinjection into HeLa cells.

2-5A[ppp(A2'p)n5'A] has been implicated as a mediator in the antiviral action of interferon. Its direct evaluation as an indicator of virus replication is hampered by two limitations: its inability to penetrate intact cells, and its rapid intracellular degradation by (2'-5')phosphodiesterase. These problems could be overcome by using a microinjection technique whereby a phosphodiesterase-resistant analog of 2-A, in which the 2'-terminals adenosine residue is replaced by 2-(9-adenyl)-6-hydroxy-methyl-4-hexylmorpholine, was injected into individual HeLa cells before infection with mengovirus or vesicular stomatitis virus (VSV). This comparative assay with two representatives of different virus classes in a single experimental system pointed to the high sensitivity of VSV to inhibition by 2-5A oligonucleotides, in contrast with the low sensitivity of mengovirus. Microinjection of the hexylmorpholine 2-5A analog led to a much greater reduction in mengovirus yield than did microinjection of 2-5A itself.

Adenine Nucleotides↗

PubMed1993

Microinjection of acylphosphatase blocks Xenopus laevis oocytes maturation induced by ras-p21.

Ras proteins induce germinal vesicle breakdown (GVBD) when microinjected into Xenopus laevis oocytes. The mechanism of action is still unresolved, although several hypotheses have been proposed. Acylphosphatase is a cytosolic enzyme that specifically catalyses the hydrolysis of the carboxylphosphate bond of acylphosphate for the removal of acylphosphate residues of various membrane pumps. A direct effect of acylphosphatase on the regulation of ionic balance of a cell by interaction with ionic membrane pumps has been proposed. We have analyzed the effect of microinjecting acylphosphatase, by itself or along with ras-p21 proteins or progesterone, into oocytes. The enzyme alone is unable to induce GVBD, but increases oocyte maturation induced by progesterone. By contrast, acylphosphatase blocked GVBD induced by microinjection of oncogenic ras-p21. These data suggest that acylphosphatase acts synergistically or antagonistically with factors involved in proliferating signals by altering the intracellular ionic conditions of the cell, conforming the hypothesis that the intracellular ionic condition of the cell is important in the induction of proliferating signals, and that its perturbation may have a serious effect on signal transduction.

Acid Anhydride Hydrolases↗

PubMed1990

Cytoplasmic microinjection of immunoglobulin Gs recognizing RNA helices inhibits human cell growth.

We report here that nucleolar and cytoplasmic RNA in mammalian cells is recognized specifically by both experimentally induced monoclonal IgG unique for left-handed Z-RNA and by autoimmune mouse monoclonal IgG specific for ribosomal RNA. Nucleolar Z-RNA synthesis, like nucleolar ribosomal RNA synthesis, is inhibited by actinomycin D treatment and dimethylsulfoxide-induced differentiation. Immune anti-Z-RNA IgGs microinjected into living nuclei bind nucleolar RNA, and these complexes appear to be removed from the nucleus within minutes. Cytoplasmically microinjected monoclonal or polyclonal anti-Z-RNA IgGs specifically bind cytoplasmic RNA and inhibit cell multiplication. Microinjection of antibodies directed against double-stranded RNAs. Elevated ionic conditions, which in energy-minimized models can cause the walls of the groove in Z-RNA (but not Z-DNA) to approach each other and close, also prevent antibody binding to specific synthetic or cellular Z-RNA determinants. Our antibodies binding unique Z-RNA structures probably recognize antigens determined by the exposed 2'-OH ribose sugar-phosphate groups.

Animals↗

PubMed1987

Microinjection of oxytocin into limbic-mesolimbic brain structures disrupts heroin self-administration behavior: a receptor-mediated event?

The systemic injection of oxytocin (OXT) decreases the self-administration of heroin in heroin-tolerant rats. Since OXT-ergic binding sites are present in limbic and mesolimbic brain regions, the effects of intracerebral microinjections of OXT were investigated. In heroin-tolerant rats, the microinjection of OXT (2 ng) into the anterodorsal part of the nucleus accumbens or into the ventral hippocampus disrupted the self-administration of heroin. The effect of intrahippocampal microinjections lasted longer than that of intraaccumbens injections. The administration of N alpha-acetyl-(2-0-methyltyrosine)-oxytocin (ACME-OXT), an inhibitor of oxytocin receptors, prevented the disruptive effect of intrahippocampal OXT injections on heroin self-administration. It is concluded that limbic-mesolimbic brain structures have an essential role in the expression of the disruptive action of OXT on heroin self-administration. It appears that OXT-ergic binding sites mediate the effects of OXT.

Animals↗

PubMed1991

Gastric effects of thyrotropin-releasing hormone microinjected into the dorsal vagal nucleus in cats.

We investigated the gastric acid secretory and motility responses to microinjection of thyrotropin-releasing hormone (TRH) into the dorsal motor nucleus of the vagus (DMV) in anesthetized cats. Gastric acid output was collected every 15 min through a gastric cannula after saline flush and titrated to pH 7.0. Antral and corpus contractions were continuously recorded by extraluminal force transducers. TRH dissolved in 200 nl of saline and microinjected unilaterally into the DMV induced a dose-dependent (50-200 ng) increase in gastric acid secretion. The acid secretory response began in the first 15 min collection and lasted 45 min. TRH frequently increased the force of contractions of the antrum and corpus within one minute of microinjection. The minimal effective dose for eliciting increased motility was lower than for inducing acid secretion. These results demonstrate that TRH acts in the DMV of cats to stimulate gastric acid secretion and contractions.

Animals↗

PubMed1992

Characterization of the hypnotic effects of triazolam microinjections into the medial preoptic area.

We have previously reported that microinjections of the benzodiazepine hypnotic triazolam into the medial preoptic area (MPA) of the hypothalamus enhance sleep in rats. The present study further characterizes this effect, by examining its anatomical specificity, determining whether it is mediated by interaction with central benzodiazepine receptors, and assessing whether sleep induction is associated with changes in core temperature. It was found that microinjections of 0.25 and 0.5 micrograms triazolam into two nearby structures, the lateral preoptic area and diagonal band of Broca, failed to alter sleep. Total sleep time was enhanced by microinjection of triazolam into the MPA, and this effect was blocked by co-administration of the benzodiazepine receptor blocker RO 15-1788. Sleep enhancement by triazolam was not associated with significant alterations in core body temperature. These observations continue to suggest that the MPA may be a site which mediates the hypnotic effect of triazolam, and add to the growing body of data emphasizing the importance of hypothalamic function in the regulation of sleep and waking.

Body Temperature↗

PubMed1986

Microinjection of GABA antagonists into posterior hypothalamus elevates heart rate in anesthetized rats.

Microinjection of the GABA antagonist bicuculline methiodide 1-25 ng into the posterior hypothalamus of urethane-anesthetized rats evoked sympathetically-mediated increases in heart rate of up to 150 beats/min and modest increases in blood pressure which could be prevented by prior local microinjection of muscimol 50 ng. Microinjection of picrotoxin but not strychnine produced similar effects. These results suggest that a latent sympathoexcitatory mechanism in this region is tonically inhibited by endogenous GABA.

Anesthesia↗

PubMed1994

Asymmetry in behavioral responses to cholecystokinin microinjected into rat nucleus accumbens and amygdala.

The behavioral responses of rats to uni- or bilateral microinjections of the octapeptide cholecystokinin (CCK-8) into the left and/or right or both nucleus accumbens (NA) or amygdalae were studied. There were two main findings of effects of microinjections of CCK-8 into NA. First, bilateral injections of CCK-8 into NA dose-dependently decreased the horizontal activity. The second more important finding was that CCK-8 at a specific dose (0.01 micrograms) injected into the right NA increased the number of horizontal movements 6-fold as compared to the injection into the left NA. Neither uni- nor bilateral injections of CCK-8 into NA at all doses used induced changes in the vertical movements. CCK-8 injected into left, right or both amigdalae increased locomotion at the lowest dose (0.01 microgram), while at the high doses (0.5 and 1.0 microgram) it significantly decreased it. The plus-maze test confirmed the anxiogenic effect of CCK-8 (0.01 microgram) injected into amigdalae. CCK-8 exerted a favorable effect on learning and memory (shuttle-box) when injected into the left but not into the right amygdala. Injection of CCK-8 (0.01 micrograms) into left amygdala provoked a 4-fold increase of the number of avoidances as compared to the microinjection into the right amygdala.

Amygdala↗

PubMed1996

Galanin microinjected into the medial preoptic nucleus facilitates female- and male-typical sexual behaviors in the female rat.

Galanin (GAL) microinjected within the sexually dimorphic medial preoptic nucleus (MPN) facilitates male-typical sexual behaviors in the male rat, a response that requires the presence of testosterone. As in the male, GAL-immunoreactive cells located within the MPN of the female also concentrate gonadal steroids and become less immunoreactive after gonadectomy. Thus, to investigate sexual behaviors in the female and to determine whether effects are comparable to those obtained in the male, GAL was microinjected unilaterally within the MPN of female rats. We report that GAL stimulated female-typical lordosis behavior after estrogen priming, and that the effect was not due to general arousal as measured by nonspecific locomotor activities. In a separate experiment, GAL microinjected within the MPN dose-responsively increased mount frequencies and decreased mount latencies in testosterone-primed females. A higher dose of testosterone was required in females for this stimulation of male-typical sexual behavior than required in a previous experiment in males.

Animals↗

PubMed1987

Effect of microinjected amine and diamine oxidases on the ultrastructure of eukaryotic cultured cells.

Diamine oxide and serum amine oxidase, which catalyse the oxidation of diamines and polyamines, respectively, were trapped within reconstituted Sendai virus envelopes. These loaded envelopes were incubated with cultured normal chick fibroblasts or with fibroblasts transformed by Rous sarcoma viruses. The binding of the reconstituted envelopes to the cultured cells was confirmed by scanning electron microscopy. It has been shown that the reconstituted envelopes (1-3 microns diameter) were attached to the eukaryotic cells. No significant changes in the morphology of the normal chick embryo fibroblasts were noted upon treatment with enzyme-loaded envelopes. On the other hand, chick embryo fibroblasts transformed by Rous sarcoma virus were affected by the microinjected amine oxidases. Scanning electron microscopy demonstrated the formation of holes in the microinjected cells. Similar morphological changes were also observed when diamine oxidase was microinjected into cultured glioma cells. These holes may be the result of the ejection of the nucleus. These findings are in line with the observed effect of the injected amine oxidases on macromolecular synthesis in normal and transformed chick embryo fibroblasts.

Amine Oxidase (Copper-Containing)↗

PubMed1987

Fate of cadmium in rat renal tubules: a microinjection study.

109Cd was injected into the lumen of superficial proximal or distal tubules of rat kidneys, and recovery in the pelvic urine from the ipsilateral kidney was measured. Fractional recovery of labeled inulin always exceeded 90%. About 70% of injected inorganic Cd (CdCl2) was taken up by the epithelium of proximal tubules, while more than 90% of the injected amount was recovered after distal microinjection. The proximal fractional Cd uptake of a 1:1 (molar) Cd-L-cysteine complex was 82%, but was below 60% for a 5-10:1 molar ratio of cysteine:Cd. The chelate Cd-pentetic acid was recovered in final urine nearly quantitatively after proximal or distal microinjection. Fractional uptake of 109Cd from a Cd-metallothionein (Mt) complex, following proximal microinjection, ranged between 17 (Cd-Mt 0.19 mM) and 8% (Cd-Mt 1.5 mM). It is concluded that luminal Cd uptake by the tubular epithelium depends markedly on the chemical form of Cd and, when present, occurs mostly or exclusively in proximal tubules.

Animals↗

PubMed1990

Effects of perifornical hypothalamic microinjections of phenylpropanolamine and amphetamine on latency to feed and mash intake in rats.

To determine whether phenylpropanolamine (PPA) and amphetamine act on a common satiety mechanism, the present experiment compared the effect of unilateral microinjections (40, 80, and 160 nmol) of phenylpropanolamine hydrochloride (PPA: d,1-norephedrine) and of d-amphetamine sulfate within the perifornical hypothalamus (PFH) on consumption of a palatable sweetened-mash diet in adult male rats. Microinjection of d-amphetamine (40-160 nmol) within the PFH induced dose-dependent anorexia, whereas PPA microinjections were without effect on feeding. These results document that amphetamine and PPA do not act at a common CNS site, such as the PFH, to induce anorexia.

Animals↗

PubMed1992

Microinjections of dopamine agonists in the nucleus accumbens increase ethanol-reinforced responding.

Long-Evans rats (N = 3) were trained to lever press on a fixed-ratio 4 (FR 4) schedule with ethanol (10% v/v) presented as the reinforcer. Each rat received a total of six bilateral nucleus accumbens microinjections, one per week. They were tested with one physiological saline control, three 20.0-microgram/brain d-amphetamine, and two 6.0-microgram/brain quinpirole injections given 10 min prior to operant sessions. Ethanol-reinforced responding terminated after approximately 10 min during control sessions. Microinjections of the D2 agonist quinpirole and the nonspecific dopamine (DA) agonist d-amphetamine increased total responding but produced slowed response rates that continued for 45-60 min. The slowed response rate produced by d-amphetamine resulted in a peak increase in interresponse times (IRTs) between 8-10 s, whereas quinpirole increased IRTs in the 14- to 16-s range, indicating that nonspecific DA activation resulted in higher rates of ethanol-reinforced responding than specific D2 activation although both drugs decreased local response rates. These data indicate that the amount and temporal extent of ethanol-reinforced responding are increased by microinjections of DA agonists in the nucleus accumbens and support the hypothesis that DA activity in this region is involved in the regulation of ethanol-reinforced responding.

Animals↗

PubMed1981

Disruption of the in vivo distribution of the intermediate filaments in fibroblasts through the microinjection of a specific monoclonal antibody.

Monoclonal antibodies (JLB1 and JLB7) that recognize minor components of the intermediate filament system of cultured cells were introduced into living fibroblasts by microinjection. Several minutes after injection of the JLB7 antibody virtually all of the intermediate filaments of the cells were found to be aggregated into tight bundles near or around the nucleus. In contrast, injection of the JLB1 antibody caused little or no aggregation of the intermediate filaments. Electron microscopy showed that the perinuclear bundles that formed after injection of the JLB7 antibody each consisted of ten or more filaments apparently crosslinked together. Double-label immunofluorescence microscopy showed that virtually all of the vimentin-containing intermediate filaments in the JLB7 antibody-injected cells were redistributed to the perinuclear region and remained there for at least 24 hr. The distributions of actin microfilaments and microtubules were seemingly undisturbed following microinjection. No obvious changes in cell morphology or behavior were apparent in the cells injected with JLB7 antibody; the cells displayed a flat appearance, showed a polarity, were able to ruffle and bleb and even appeared to show the normal saltatory movements of intracellular vesicles, granules and mitochondria, suggesting that intermediate filaments are not involved in these activities. The microinjection of highly specific monoclonal antibodies that recognize and alter components of the cell provides an additional approach to determine the in vivo functions of intracellular elements.

Animals↗

PubMed1985

Microinjection of the ras oncogene protein into PC12 cells induces morphological differentiation.

To investigate the possible role of ras proteins in the differentiation process signaled by nerve growth factor, we have microinjected the proto-oncogenic and oncogenic (T24) forms of the human H-ras protein into living rat pheochromocytoma cells (PC12). PC12 cells, which have the phenotype of replicating chromaffin-like cells under normal growth conditions, respond to nerve growth factor by differentiating into nonreplicating sympathetic neuron-like cells. Microinjection of the ras oncogene protein promoted the morphological differentiation of PC12 cells into neuron-like cells. In contrast, microinjection of similar amounts of the proto-oncogene form of the ras protein had no apparent effect on PC12 cells. The induction of morphological differentiation by the ras oncogene protein occurred in the absence of nerve growth factor, was dependent on protein synthesis, and was accompanied by cessation of cell division. Treatment of PC12 cells with nerve growth factor or cAMP analogue prior to injection did not alter the phenotypic changes induced by the ras oncogene protein.

Adrenal Gland Neoplasms↗