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At least 775 records · Page 43Linked to original sources

PubMed1990

Sensitivity of the sodium efflux in single barnacle muscle fibers to the microinjection of aluminum.

A study has been made of the behavior of the Na efflux in single muscle fibers from the barnacle, Balanus nubilus, toward the microinjection of AlCl3. The effect of microinjecting AlCl3 is either biphasic with inhibition following transitory stimulation, or monophasic with inhibition occurring promptly and taking about 1 hr to reach a maximum. The magnitude of the inhibition is dose dependent and the minimally effective concentration is 10 mM. This is diluted by the myoplasm by a factor of roughly 100. Fibers injected with 1 M AlCl3 are found to be insensitive to 10(-4) M ouabain, whereas fibers injected with 0.5 M AlCl3 show a further fall in Na efflux, which is smaller than that seen in control fibers. The response of the ouabain-insensitive Na efflux to the injection of 0.5 M AlCl3 is biphasic: stimulation is followed by inhibition. The stimulatory phase is largely dependent on the presence of external Ca2+ and is reversed by the sudden omission of Ca2+ from the bathing medium. Fibers also injected with 0.5 M AlCl3 show a marked reduction in the response of the Na efflux to high external K. Injection of ATPMg before or after the injection of AlCl3 fails in most instances to abolish or reverse the inhibitory effect. By contrast, fibers preinjected with deferoxamine show little or no effect following the injection of AlCl3. However, external application of deferoxamine is ineffective.

Aluminum↗

PubMed1990

2,5-Hexanedione-treated tubulin microinjected into sea urchin zygotes induces mitotic abnormalities.

Zygotes of Lytechinus pictus and Lytechinus variegatus were microinjected with 2,5-hexanedione (2,5-HD)-treated tubulin prior to the first mitotic cycle. Mitotic spindles were small with a well-defined metaphase plate, but poor birefringence and poor astral development. Abnormalities were observed in chromosome movement at anaphase and cytokinesis. Neither microinjections of untreated tubulin or 3-acetyl-2,5-hexanedione-treated tubulin, nor incubation of zygotes in 2,5-HD-containing sea water produced abnormalities. The results can be explained in terms of the nondissociating properties of 2,5-HD-treated tubulin. 2,5-HD-treated tubulin dissociates slowly from microtubules, a property which, besides favoring the formation of stable microtubules, allows this tubulin to induce microtubule assembly when present in substoichiometric amounts. These characteristics have been implicated as a cause of 2,5-HD-induced Sertoli cell dysfunction. The effect of 2,5-HD-treated tubulin on microtubule dynamics in sea urchin zygotes may bear similarities to the effects of 2,5-HD treatment in vivo on Sertoli cell microtubules.

Animals↗

PubMed1995

A microinjection technique using Drosophila melanogaster for bioassay-guided isolation of neurotoxins in arthropod venoms.

Modern analytical techniques permit isolation and structural determination of neurotoxins at the picomole level. However, bioassay-guided fractionation of the sample often relies on simple injection assays using insects, vertebrates or crustaceans of a fairly large size, thus consuming quite a large amount of the samples being investigated. In order to investigate samples of very small size, we have devised an insect microinjection method using glass micropipettes and Drosophila melanogaster adults as test insects. The validity of the method was tested with a series of six buthoid scorpion venoms (Androctonus australis, Buthotus judaicus, Buthus tamulus, Centruroides sculpturatus, Leiurus quinquestriatus hebraeus, Tityus serrulatus) and one chactoid scorpion (Scorpio maurus palmatus) as standards. The LD50S of the venoms were determined using both the microinjection method and a classical injection assay with crickets (Gryllus bimaculatus) as test insects. Results demonstrated that the new method can successfully be applied to the study of insect neurotoxic activity in arthropod venoms. The Gryllus:Drosophila ratio in amount of sample utilized is 100. However, for all Buthoid venoms tested, except L. quinquestriatus, Drosophila showed less sensitivity, thus reducing the gain by a factor of 2-10. Drosophila were several times more sensitive to the only chactoid venom tested. These results clearly demonstrate the advantage of using this microtechnique, when limited amounts of material are available for both chemical and biological work.

Animals↗

PubMed1991

Microinjected Coxsackie B1 virus does not replicate in HEp-2 cells.

Coxsackie B1 virus did not replicate when microinjected into HEp-2 cells. Replication was assayed by production of infectious virus particles, synthesis of viral RNA, and lysis of cells. The same virus preparation initiated replication when it was inoculated into HEp-2 cells similarly to that by microinjected purified coxsackie B1 virus RNA.

Cell Death↗

PubMed1988

ATP-gamma-S (adenosine 5'-O-(3-thiotriphosphate)) microinjection increases progesterone-stimulated histone kinase activity in Xenopus oocytes.

Progesterone stimulates the activity of a potent Ca2+- and cyclic nucleotide-independent histone kinase in Xenopus oocytes. Microinjection of ATP-gamma-S during the course of meiotic maturation or after the oocytes had arrested at second meiotic metaphase resulted in a further increase of this histone kinase activity in progesterone-stimulated oocytes. ATP-gamma-S microinjection also increased and stabilized the activity of the maturation-promoting factor. The possible mechanism of ATP-gamma-S action is discussed.

Adenosine Triphosphate↗

PubMed1987

Red cell-mediated microinjection.

Red cell-mediated microinjection facilitates the rapid introduction of macromolecules into large numbers of culture cells. Efficient injection of cells in monolayer using PEG as fusogen can be accomplished using any of three alternative agglutinating agents (PHA, Con A, or polylysine) to attach loaded red cells to recipient cells. These three method yield similar efficiencies of microinjection and postfusion viability, as shown in Table I. Either of the methods, after being tailored to fit particular needs, can be expected to produce results similar to those in Table I with some variability due to differences in cell types. One drawback when PHA is used as agglutinin is the inability to remove adherent, nonfused red cells from cultures after fusion. Reversible attachment of red cells is possible using Con A or polylysine. For each of the latter agglutinins, reversal of binding detaches the great majority of the red cells; virtually all nonfused red cells may be eliminated by performing an additional simple step in either case.

Agglutinins↗

PubMed1977

A simple chronic microinjection system for use with chemitrodes.

An inexpensive, electrically operated microinjection system is described which is small enough to be mounted on the animal side of a standard Mercury commutator. This eliminates the troublesome swivel joint needed for chronic microinjection in freely moving animals. This system allows simultaneous electrical stimulation or recording and chemical stimulation via a chemitrode using a standard 4-channel Mercury commutator.

Animals↗

PubMed1981

Modification of motor activity, passive avoidance conditioning and evoked potentials by microinjections of strychnine in both caudate nuclei in cats.

Passive avoidance conditioning (PAC) seems to depend on inhibitory actions within the caudate nucleus (CN). Thus topical application of strychnine could block these influences. In cats, with permanently implanted cannulae in the head of the CN, bilateral microinjections of 100 micrograms of strychnine produced a stereotyped behavior of grooming and running with the extremities flexed to dark places. When placed into a two compartment chamber for a PAC acquisition they would not move from the dark compartment to the illuminated one. After decreasing the illumination some cats crossed and the acquisition test was performed. Twenty four hours later they showed the learned response. A second retention test was not modified by another strychnine application. These results contrast with those obtained by picrotoxin microinjections which disrupt the retention of a PAC. However, strychnine affects the evoked potentials (EP) recorded in CN by n. centralis medialis (NCM) stimulation by decreasing a positive peak, which probably represents post-synaptic inhibition and increasing the late positive component. In the lateral geniculate body strychnine affected a late positive wave and in the occipital cortex it increased the initial negative peaks and decreased the late positive ones of the potentials evoked by flash stimulation. These findings suggest a more suitable role of the CN in the regulation of visual information, which is probably related with the attention processes.

Animals↗

PubMed1981

Analgesia following microinjection of phosphodiesterase inhibitors at brainstem sites.

Microinjection of RO 20-1724 (0.60 micrograms) at sites in the caudal reticular formation of the rat brainstem resulted in analgesia as measured in the tail flick, paw pinch and hot plate tests. Microinjection of theophylline (0.45 micrograms) at sites in the caudal reticular formation also produced analgesia in the tail flick and hot plate tests, but not in the paw pinch test. Since these agents inhibit phosphodiesterase and elevate cyclic nucleotides in rat brain homogenates, these data suggest that cyclic nucleotides have a functional role in brainstem structures involved in modulation of nociceptive threshold.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗

PubMed1983

Interaction between noradrenergic and serotonergic brain systems as evidenced by behavioral and biochemical effects of microinjections of adrenergic agonists and antagonists into the median raphe nucleus.

The effects of microinjections of adrenergic receptors agonists and antagonists into the median raphe nucleus (MR) on behavior and serotonin (5HT) metabolism was examined in rats. Administration of adrenergic alpha 1 and alpha 2 receptor agonists (noradrenaline, phenylephrine, clonidine) produced behavioral excitation in the open field test and a tendency to decrease the forebrain 5-hydroxyindolo-acetic acid (5HIAA) concentration. Opposite effects were seen after microinjection of adrenergic alpha receptor antagonists (phenoxybenzamine, phentolamine but not yohimbine). A significant negative correlation was found between the effects on locomotor activity and 5HIAA levels in these rats. No effect was present after injection of beta receptor agonist salbutamol or antagonist propranolol. It is suggested that noradrenaline released from noradrenergic terminals in the MR tonically inhibits the activity of 5HT neurons thus producing symptoms of 5HT deficiency and that this action of noradrenaline is probably limited to the effects on alpha 1 but not alpha 2 nor beta adrenoceptors in this brain region.

Adrenergic Agonists↗

PubMed1983

Antinociception following microinjection of dibutyryl cyclic nucleotides into the caudal reticular formation and periaqueductal gray of the rat brain.

The tail flick, paw pinch, and hot plate tests were used to assess changes in nociceptive threshold following microinjection of dibutyryl derivatives of cyclic nucleotides into areas of the central nervous system previously shown to be involved in modulation of nociceptive threshold and mediation of morphine analgesia. An elevation in the nociceptive threshold was observed on all three tests following administration of 10 micrograms dibutyryl cyclic 3':5' adenosine monophosphate (db cAMP) into the caudal brainstem reticular formation (CRF) and periaqueductal gray (PAG). Two micrograms db cAMP produced the same magnitude of analgesia but had a shorter duration of action. Twenty micrograms dibutyryl cyclic 3':5' guanosine monophosphate (db cGMP) produced analgesia on all three tests following microinjection at CRF sites but not at PAG sites. These data indicate that morphine analgesia and the antinociception produced by cyclic nucleotides may involve, at least in part, common neuronal substrates. However, the observed capacity of db cAMP to elevate nociceptive threshold does not support the hypothesis that the mechanism of morphine's analgesic action involves inhibition of adenylate cyclase.

Analgesia↗

PubMed1984

Self-injurious behavior in rats produced by intranigral microinjection of GABA agonists.

Bilateral injection of the GABA agonist muscimol (10-300 ng) into the caudal substantia nigra (pars reticulata) of rats produced dose-dependent stereotyped gnawing and self-biting. Limiting the opportunity to gnaw on inanimate objects shifted the dose-response curve for muscimol-induced self-injurious behavior (SIB) to the left and increased the maximum incidence of SIB. Microinjection of muscimol (30 ng) into the rostral and caudal regions of the substantia nigra were equally effective in producing SIB, though the incidence of SIB decreased sharply when muscimol was injected 1 mm rostral or caudal to the substantia nigra. Bilateral intranigral injection of THIP (100-1000 ng) and (+/-)baclofen (100-1000 ng) induced a low incidence of SIB. However, neither IP administration of picrotoxin (5 mg/kg) or simultaneous microinjection of (+)bicuculline methiodide (BMI; 300 or 1000 ng) along with muscimol (30 ng) blocked muscimol-induced SIB. In fact, (+)BMI increased the occurrence of self-biting and reduced the latency to onset of SIB. The involvement of GABAergic mechanisms in muscimol-induced SIB is discussed.

Animals↗

PubMed1984

Comparison of the CNS effects induced by TRH and bicuculline after microinjection into medial septum, substantia nigra and inferior colliculus: absence of support for a GABA antagonist action for TRH.

Antagonism of ethanol-induced depression of locomotion was observed after intracisternal injection of thyrotropin releasing hormone (TRH) and bicuculline methiodide (BICM), as well as after microinjection of these drugs into the medial septum. The present investigation compared the behavioral and physiological consequence of administering TRH and BICM into the medial septum, inferior colliculus and substantia nigra to quantitate the similarities between these compounds. BICM produced a major increase in locomotor activity when injected into the medial septum and stereotypies when injected into the substantia nigra, suggesting that GABA-containing neurons have widespread influences on motor function. The wild running and seizure activity observed after BICM injection into the inferior colliculus was also consistent with this latter view. The marked increase in rectal temperature observed when BICM was injected into the medial septum may also implicate GABAergic mechanisms in temperature control at this brain site. TRH produced no such behavioral or physiological changes when administered into these three sites. Thus, this work strongly suggests that TRH does not exert a widespread action as a GABA antagonist because TRH did not produce the same changes induced by BICM. The actions of BICM and TRH to antagonize ethanol-induced depression when microinjected into the medial septum suggests that this brain area may be a critical site for the depressant action of ethanol.

Animals↗

PubMed1995

Behavioural effects induced by microinjection of L-BOAA into the ventrolateral PAG matter of the mouse.

L-BOAA (1 microgram/mouse), microinjected into the ventrolateral periaqueductal gray (PAG) matter, induced a strong reaction of forward avoidance (running) for 20-30 s in 18% of the mice and immobility for 7 +/- 2 min in 72% of the mice from a total of 68 treated animals. Effects were also observed for grooming and clonus in 6% and 4% of the mice, respectively. Duration of L-BOAA-induced immobility was significantly (p < 0.05) reduced by a pretreatment with CNQX (0.5 microgram/mouse), a selective antagonist of AMPA glutamergic subtype receptors, but not by a pretreatment with 2-APV (0.5 microgram/mouse), a selective antagonist of NMDA glutamergic subtype receptors, nor by 2-AP3 (0.5 microgram/mouse), a weak antagonist of metabotropic glutamergic subtype receptors. AMPA (0.05 microgram/mouse), also microinjected into ventrolateral PAG, induced the same pattern of behavioural effects as L-BOAA. Forward avoidance, grooming, and clonus induced by L-BOAA or AMPA were also significantly antagonised by a pretreatment with CNQX (data not shown).

Amino Acids, Diamino↗

PubMed1975

Microinjection of thymidine kinase and bovine serum albumin into mammalian cells by fusion with red blood cells.

A procedure is described by which proteins can be rapidly and efficiently microinjected into large numbers of culture cells. Proteins were first introduced into mammalian red blood cells during hypotonic hemolysis, and the resealed red cells were subsequently fused to culture cells using Sendai virus. In seven experiments, thymidine kinase or 125I-BSA were transferred to culture cells during fusion. Although proteins were used in the present experiments, the microinjection procedure should work equally well for other macromolecules.

Animals↗

PubMed1976

Microinjection studies of duck globin messenger RNA translation in human and avian cells.

Microinjection techniques were used for the study of the translation of heterospecific messenger RNAs in living cells. Duck globin messenger RNAs, in the form of polysomes, messenger ribonucleoprotein particles, or free mRNAs, were injected into cells of human origin (HeLa), and the subsequent appearance and accumulation of avian globin were measured by specific immunofluorescence techniques. Globin accumulated within a high proportion of HeLa cells for at least 25 hr following injection, indicating the intracellular stability of the heterospecific globin messenger RNA and the fact that mRNA initially associated with more complex structures could be translated in the HeLa cytoplasm. In similar experiments, duck reticulocyte polysomes were shown to be translated in duck embryo cells but not in embryonic chick liver cells. The latter, however, could be shown to withstand the microinjection procedure as judged by their capacity to respond normally to inducers and inhibitors of the synthesis of alpha-aminolevulinic acid synthetase. HeLa cells injected with duck hemoglobin divide at normal or nearly normal rates following the injection, but their globin content diminishes, in contrast with the increasing globin content of cells receiving duck reticulocyte polysomes, mRNP particles, or free mRNA.

5-Aminolevulinate Synthetase↗

PubMed1980

High efficiency transformation by direct microinjection of DNA into cultured mammalian cells.

Direct microinjection of DNA by glass micropipettes was used to introduce the Herpes simplex virus thymidine kinase gene into cultured mammalian cells. When DNA was delivered directly into the nuclei of LMTK-, a mouse cell line deficient in thymidine kinase activity, 50--100% of the cells expressed TK enzymatic activity. In contrast, no TK activity could be detected when the DNA was injected into the cytoplasm. The number of injected LMTK- cells capable of indefinite growth in a TK+ selective medium (that is, transformants) depended on the nature of the plasmid DNA into which the HSV-TK gene was inserted. One cell in 500-1000 cells which received nuclear injections with pBR322/TK DNA gave rise to a viable colony when grown in HAT medium (that is, a TK+ selective medium). The transformation frequency increased to one in five injected cells when specific SV40 DNA sequences were also introduced into the HSV-TK plasmid. With the microinjection procedure transformation frequency was relatively insensitive to DNA concentration and did not depend on co-injecting with a carrier DNA. Most of the transformants were stable in nonselective medium as soon as they could be tested.

Cell Nucleus↗

PubMed1981

Microinjection of anticlathrin antibodies into fibroblasts does not interfere with the receptor-mediated endocytosis of alpha2-macroglobulin.

Affinity-purified antibodies prepared against the major coat protein of brain coated vesicles, clathrin, were microinjected into cultured fibroblasts, and their intracellular distribution was followed by immunofluorescence microscopy and ultrastructural immunocytochemistry. Microinjected anticlathrin antibodies were concentrated on coated regions of the plasma membrane and the GERL apparatus. When an excess of anticlathrin antibodies was injected into the cytosol, coated pits on the plasma membrane were covered by anticlathrin antibody but still functioned to cluster an internalize alpha2-macroglobulin. These results are discussed in terms of the role of clathrin in the pathway of receptor-mediated endocytosis. Our data indicate that in cultured fibroblasts coated pits are stable elements permanently attached to the plasma membrane.

Animals↗