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Results for “Microinjection”

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At least 631 records · Page 35Linked to original sources

PubMed1990

Microinjection of cocaine into the nucleus accumbens elicits locomotor activation in the rat.

Cocaine is believed to exert its psychostimulant effects through activation of the mesocorticolimbic system. Although the nucleus accumbens, in particular, has been hypothesized as the site of action of cocaine's stimulating effects, there is no direct evidence that microinjection of cocaine into this region produces behavioral activation. The present experiments investigated the locomotor response to microinjection of cocaine (0, 10, 30, 100 micrograms/0.5 microliter) into the nucleus accumbens in rats. Cocaine elicited a pronounced, dose-dependent motor activation of approximately 60 min duration. This stimulant effect was blocked by prior administration of a dopamine (DA) receptor antagonist, cis-flupenthixol. The response to cocaine was differentiated from nucleus accumbens microinjections of procaine and lidocaine, compounds that have potent local anesthetic effects but little affinity for the dopamine-uptake site. Neither procaine nor lidocaine (0, 10, 30, 100 micrograms/0.5 microliter) had any overall effect, although activity was somewhat decreased in the initial part of the test session and increased at the end, relative to control activity. Cocaine injected into the anterior dorsal or ventrolateral striatum (100 micrograms) also increased motor activity; procaine and lidocaine had no effect. Cocaine injected into the ventrolateral striatum significantly increased stereotypy. The amplitude of motor activation following cocaine injection into nucleus accumbens was much greater than that elicited at the other striatal sites. Further, observation of the time course of motor activation following cocaine injection into the anterior dorsal and ventrolateral striatum suggested that the motor effect was due to diffusion, most likely to the nucleus accumbens.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1989

Cardiovascular responses of rats to electrical stimulation of the fastigial nucleus and local microinjection of amino acids.

The cardiovascular response to electrical stimulation of the fastigial nucleus (FN) and microinjection of L-glutamate, kainic acid, glycine and GABA into the FN of anesthetized rats were studied. Electrical stimulation of FN produced pressor or depressor responses with a decrease or no change in heart rate depending upon the intensity or site of the stimulation. Microinjection of L-glutamate or kainic acid into the FN elicited pressor or depressor response with bradycardia, while that of glycine or GABA did not affect blood pressure or heart rate. L-glutamate or kainic acid microinjected into the FN did not affect epinephrine-induced pressor response, but attenuated the bradycardic response. These results indicate that L-glutamate in the FN may play a role in modulation of the cardiovascular regulation.

Amino Acids↗

PubMedDate not supplied

[Biological effects accompanying the microinjection of adenovirions, adenoviral and plasmid DNA into mammalian cells].

Microinjection of either type 1 human adenovirus, type SA7 monkey adenovirus virions or circular adenovirus DNA, obtained by the treatment of DNA-terminal protein complexes with glutaraldehyde, into nuclei of permissive cells results in the complete cycle of virus reproduction. Microinjection of neither linear native, condensed adenovirus DNA nor the DNA-terminal protein complexes under the same conditions initiates the adenovirus reproduction thought the synthesis of early and some late viral antigens is observed in the injected cells. Integration of injected adenovirus DNA into the cellular DNA occurs as far as 30 min after injection. Microinjection of either adenovirus DNA or its oncogene containing fragments into nuclei of semipermissive cells induces the transformation of these cells. In this case the time of the first appearance of transformation foci is decreased.

Adenoviridae↗

PubMed1989

Mechanism of chromosomal integration of microinjected DNA.

Most transgenic animals have been produced by directly injecting DNA into one of the embryo pronuclei in the period immediately following fertilization. Transgenic animals are produced when DNA becomes integrated into the chromosomes, in most cases the transgenic genotype is transmitted to progeny through the germ line. The characteristics of the foreign DNA include tandem arrangement of multiple copies of the input DNA, predominantly in direct rather than inverted orientation, and illegitimate recombination with the chromosome at the site of integration. Foreign DNA integrated into chromosomes of cultured cells has identical characteristics. We argue that these and other similarities between the integration of foreign DNA into the chromosomes of microinjected embryos on the one hand and on the other of transfected and microinjected cells, strongly suggest that the processes have the same basis. By considering mainly the literature relating to cell transfection and microinjection, we conclude that tandemly arranged concatemers of input foreign DNA are built up by a process of homologous but non-conservative recombination. End-joining and illegitimate recombination events characterize the integration of the concatemers into the chromosome, and also contribute to the formation of the concatemers. We also suggest that these superficially different processes are based on the same opportunistic repair-ligation mechanism; the frequencies of the different types of event reflect both the frequencies with which different sorts of association occur between DNA molecules and their relative stability.

Animals↗

PubMed1985

Effect of butyrate on the expression of microinjected or transfected genes.

We have studied the effect of sodium n-butyrate on the expression of specific genes. For this purpose, tk-ts13 cells (a thymidine kinase-deficient mutant originating from Syrian hamster cells) were microinjected or transfected with pC2, a plasmid containing the entire SV40 genome and the herpes simplex virus thymidine kinase gene (HSV-TK), cloned in pBR322. As a measure of the expression of these two genes, one of which is spliced (SV40) and the other one (HSV-TK) which is not, we have taken the protein levels (amount of T antigen for SV40 and incorporation of [3H]thymidine for HSV-TK) and the levels of RNA (by dot blot hybridization). The expression of the microinjected genes was inhibited when tk-ts13 cells were exposed to butyrate, actinomycin D, cycloheximide, and mitomycin C, but not when the cells were treated with insulin or dexamethasone. Further studies showed that a decrease in the percentage of T-positive cells occurs at lower concentrations of butyrate than a decrease in the levels of specific mRNA. In tk-ts13 cells transfected with pC2 and treated with butyrate at a concentration of 3 mM, SV40 mRNA levels are not decreased but the percentage of T-positive cells is decreased 50%. At 5 mM, the amount of T antigen/cell is decreased a further 40%. These results indicate that butyrate may have at least two sites of action, one at the level of mRNA amount and a second at the level of protein amount. In addition, our studies show that the use of microinjected or transfected genes offers certain unique possibilities for studies on the effects of environmental manipulations on gene expression.

Antigens, Viral, Tumor↗

PubMed1988

Stimulation of exocytotic degranulation by microinjection of the ras oncogene protein into rat mast cells.

To investigate the possible role of ras proteins in the secretory process, we have microinjected the proto oncogenic and oncogenic forms of the human H-ras protein into rat peritoneal mast cells. Mast cells are secretory cells which, upon appropriate stimulus, liberate histamine and other mediators of the acute inflammatory reaction by exocytotic degranulation. We report here that microinjection of the ras oncogene protein into mast cells induces exocytotic degranulation. In contrast, microinjection of similar amounts of the proto-oncogenic protein has little apparent effect on mast cells. Degranulation induced by injection of the ras oncogene protein occurs in the absence of an external stimulus and requires the presence of external calcium. The ultrastructural features of exocytotic degranulation in mast cells injected with the ras oncogene protein are similar to those seen when mast cells are activated by soluble ligands. Our results suggest that ras proteins may be involved, possibly as regulatory elements, in cellular functions that control exocytosis.

Animals↗

PubMed1978

Red cell-mediated microinjection.

I have reviewed the current status of microinjection based on fusion of red blood cells and tissue culture cells. Macromolecules are introduced into red blood cells during hypotonic hemolysis, and the resealed red cells are then fused to tissue culture cells with Sendai virus. The procedure has been used to inject ferritin, thymidine kinase, bovine serum albumin, and transfer RNA molecules into large numbers of tissue culture cells. Physiologically significant amounts of various macromolecules can be transferred, and preliminary studies show that [125I]bovine serum albumin and transfer RNA are stable within recipient culture cells. Tissue culture cells remain viable following microinjection. Red cell-mediated microinjection should facilitate the study of various processes, such as macromolecular turnover and genetic regulation, that are not easily studied with conventional biochemical techniques.

Animals↗

PubMed1984

[Effect of bulbar microinjections of noradrenaline, isoproterenol and phentolamine on the spino-bulbo-spinal reflex in the cat].

In chloralose-anesthetized cats a long lasting depression of spino-bulbo-spinal (SBS) reflex discharges following microinjections of noradrenaline and its agonist (isoproterenol) in the medial bulbar reticular formation was revealed. Microinjections of phentolamine (alpha-adrenoceptor antagonist) were followed by prominent enhancement of SBS-reflexes. Depressive action of the locus coeruleus stimulation on SBS-responses essentially decreased after these microinjections. The data evidence for a specific noradrenergic nature of inhibitory locus coeruleus influences on reticular centre of SBS-reflexes.

Animals↗

PubMed1995

Fast inducible repair of microinjected UV-irradiated SV40 DNA in monkey kidney cells.

In monkey kidney cells (TC-7), microinjected with UV-irradiated (103-362 J/m2) SV40 DNA, the expression of viral antigens decreases in a UV-dose-dependent manner and the viral genes are not repaired constitutively. When the viral DNA is microinjected 4 h after UV-irradiation (40 J/m2) of host cells, the expression of viral antigens is restored in all cells. The time course of restoration of viral gene expression function shows that in UV-irradiated cells the repair is induced rapidly and fully within 2 h and the induced state is maintained for 24 h. Intact viral DNA molecules, microinjected during the period of induction of cellular UV repair, are expressed less efficiently than UV-irradiated viral genomes.

Animals↗

PubMed1994

[Influence of thoracic spinal subarachnoid microinjection of phentolamine on the effect of electroacupuncture].

The influence of thoracic spinal subarachnoid microinjection of Phentolamine (Phen, 200ug/20ul) on the effect of electroacupuncture (EA) at Neiguan (PC 6) was observed in 42 urethan-chloralose anesthetized rabbits. Results showed that: 1) EA of Neiguan (PC 6) could significantly promote recovery of elevated ST-segments and T-waves of electrocardiogram induced by acute myocardial ischemia (AMI). 2) subarachnoid microinjection of phentolamine had no marked influence on recovery of ST-segments and T-waves, but impeded recovery of blood pressure of post-AMI; and 3) subarachnoid microinjection of phentolamine could significantly weaken or eliminate the effect of EA of Neiguan (PC 6) in accelerating recovery of ST-segments and T-waves of post-AMI. It denotes that alpha-receptors of intra-thoracic spinal cord participate in the action of EA of Neiguan (PC 6) in improving AMI, and the upper thoracic segment of the spinal cord is one of the links for connecting Neiguan and the heart.

Animals↗

PubMed1993

Serotonin microinjected into the nucleus raphe obscurus increases intragastric pressure in the rat via a vagally mediated pathway.

The purpose of the present study was to investigate the effect of microinjection of serotonin (5-HT) and selected 5-HT receptor subtype agonists and antagonists into the caudal nucleus raphe obscurus on gastrointestinal motor activity in urethane-chloralose anesthetized rats. Serotonin (0.6-18.0 nmol) dose-dependently increased intragastric pressure, and this effect was abolished by peripherally administered atropine (0.5-1.0 mg/kg, i.v.). Microinjection of a 5-HT1A receptor agonist, 8-hydroxy-N,N-dipropyl-2-amino-tetralin hydrobromide (0.06-12.0 nmol), a 5-HT1C/2 receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl (4.5 and 18.0 nmol), as well as a 5-HT3 receptor agonist, 1-(m-chlorophenyl)-biguanide hydrochloride (0.6-18.0 nmol), also resulted in increases in intragastric pressure. The gastric excitatory effect of 5-HT (6.0 nmol) was markedly reduced by prior microinjection of a 5-HT1/2 receptor antagonist, methiothepin (200 nmol), into the same site, as well as by i.v. administration of a 5-HT2/1C antagonist, ketanserin (2.5 mg/kg). The effect of 5-HT (6.0 nmol) on intragastric pressure was completely blocked by i.v. administration of a mixture of the 5-HT1A receptor antagonist 1-(2-methoxyphenyl)-4-[-(2-phthalimido)butyl]piperazinehydrobromide++ + (3.5 mg/kg), ketanserin (2.5 mg/kg) and the 5-HT3 receptor antagonist 3-tropanyl-3,5-dichlorobenzoate (2.5 mg/kg). These results indicate that 5-HT activates gastric motor function in the caudal nucleus raphe obscurus via a vagally mediated pathway and that the activation of multiple 5-HT receptor subtypes is required for the gastric excitatory effect of 5-HT.

Animals↗

PubMed1993

Changes in sleep patterns by intralaminar thalamic microinjection of fluoxetine in rats.

Sleep patterns were investigated in rats by analysis of EEG recorded for 6 hours following fluoxetine or saline (control) microinjection into the centromedial intralaminar thalamus (CIT) at either 9:30 AM or 1:30 PM, 4 or 8 hours after lights on, respectively. The treatment of fluoxetine at 9:30 AM did not affect any sleep patterns. In contrast, fluoxetine significantly suppressed total paradoxical sleep (PS) time and the number of PS episodes during the second and third hour after administration at 1:30 PM. In addition, microinjection of fluoxetine produced an apparent enhancement of the duration of slow wave sleep (SWS) episodes and a small non-significant increase in total SWS time. On the other hand, no obvious changes in the basal sleep-wakefulness pattern were found between the two saline-treated control groups. These results indicate that CIT microinjection of fluoxetine at different time points acts differentially in sleep patterns and suggest that CIT may be one of the action sites for peripherally administered fluoxetine to alter sleep patterns.

Animals↗

PubMed1993

In vivo experimental induction of interdigital tissue chondrogenesis in the avian limb bud results in the formation of extradigits. Effects of local microinjection of staurosporine, zinc chloride and growth factors.

In previous studies we had found that at late stages of development, when the early patterning control mechanism have ceased to act, the chick limb bud is able to form fully differentiated extradigits by subjecting the interdigital spaces to ectoderm removal. In this study we attempted to mimic this phenomenon by using local microinjections of substances which presumably have a biological action on the interdigital mesenchyme. Microinjection of staurosporine results in the formation of fully differentiated extradigits. The action of this drug appears to be due to the induction of chondrogenesis after the inhibition of the protein kinase C. Zinc chloride administration also causes ectopic chondrogenesis but it seems to act by arresting the interdigital cell death program through endonuclease inhibition. A clear differentiation of the zinc-induced cartilages into extradigits was no detected. This can be explained by the accompanying damage caused by zinc in the growing limb mesenchyme as deduced by the high incidence of hypophalangy in the normal digits. Both TGF beta 1 and TGF beta 2 have a weak effect as inducers of interdigital chondrogenesis; presumably they act by inducing chondrogenetic differentiation. Neither FGF nor EGF has any effect when administered by local microinjection. These results show that ectopic interdigital chondrogenesis induced by drug administration results in the differentiation of extradigits. This suggests that once a cartilage is formed in the autopodium it triggers a new signalling stage which leads to the morphogenesis of a digit. This morphogenetic process involves the patterning of skeleton, joints and tendons. In accordance with these observations, it can be proposed that early patterning of the limb results in the establishment of an autopodium with a defined but still plastic skeletal distribution pattern, while morphogenesis of each autopodial element would take place at a second stage by the activation of new signalling processes.

Alkaloids↗

PubMed1993

Attenuation of behavioral consequences of immobilization stress by intra-hippocampal microinjection of zimelidine.

To investigate the role of hippocampal 5-HT neurotransmission on adaptation to aversive events, individually housed male Wistar rats (200-250 g) were immobilized for 2 h and tested 24 h later in an elevated plus-maze. Immediately after the restraint period they received bilateral microinjections into the dorsal hippocampus of either saline (0.5 microliters) or the nonselective 5-HT1 antagonist dl-propranolol (20 nmol/0.5 microliters). In a second experiment the first microinjection of saline or dl-propranolol was followed by a second microinjection of saline (0.5 microliters) or the 5-HT reuptake blocker zimelidine (20 nmol/0.5 microliters). Although dl-propranolol alone did not change exploration of the elevated plus-maze, it antagonized the increase in the percentage of open arm entries induced by zimelidine (26.0 +/- 4.1 vs 5.64 +/- 3.7 in controls). These results are compatible with the view that post-synaptic 5-HT1a receptors in the hippocampus mediate adaptive or coping responses to aversive events.

Animals↗

PubMed1995

[Influence of thoracic spinal subarachnoid microinjection of noradrenaline on the effect of electroacupuncture].

Influence of thoracic spinal subarachnoid microinjection of noradrenaline (NA, 40 mg/20 microliters) on the effect of electroacupuncture (EA) at Neiguan (PC6) was observed in 40 rabbits anesthetized with urethan-chloralose. Results showed that: 1) EA of Neiguan (PC6) could significantly accelerate the recovery of ST-segment and T-wave of electrocardiogram induced by acute myocardial ischemia (AMI); 2). Subarachnoid microinjection of NA also could promote the recovery of ST, T and blood pressure of post-AMI; and 3) Subarachnoid microinjection of NA could strengthen the effect of EA of Neiguan (PC6) to promote the recovery of ST and T of Post-AMI. It suggests that alpha-receptors of intra-thoracic spinal cord participate in the action of EA of Neiguan (PC6) ameliorating AMI, and the upper thoracic segement is one of the links for connecting Neiguan and the heart.

Acupuncture Therapy↗

PubMed1995

[Characteristics of the effect of microinjections of scopolamine into the rat neostriatum on the realization of a conditioned food reflex with different degrees of reinforcement].

In chronic experiments in 32 male Sprague-Dawley rats on the model of food instrumental reflex (Skinner box) it was shown that the effect of Neostriatal scopolamine microinjections (0.3 mkg) depended on the stage of reflex consolidation. Before the complete reflex consolidation the bilateral microinjections of scopolamine into Nucleus Caudatus produce prolonged inhibition of instrumental reflex. Bilateral Neostriatal microinjections of scopolamine in the same doze had no effect in the case of full consolidation of instrumental reflex. Our results suggest that the cholinergic Neostriatal system is crucially involved in the forming of motor engram of instrumental reaction. In the same time the cholinergic neostriatal system either is not involved in the execution of full consolidate instrumental reactions, or another different forebrain structures patricipate in their realization compensating disturbances of striatal cholinergic function.

Animals↗

PubMed1997

Self-injurious behavior and dopaminergic neuron system in neonatal 6-hydroxydopamine-lesioned rat: 2. Intracerebral microinjection of dopamine agonists and antagonists.

Intracisternal 6-hydroxydopamine treatment to newborn rats caused massive and permanent damage of brain dopaminergic neurons, and many of these animals show self-injurious behavior (SIB) when loaded by systemic injection of L-dihydroxyphenuylalanine (L-DOPA) or D1 agonist, SKF-38393. SIB occurred at life-long time in neonatal 6-hydroxydopamine-lesioned rats, because SIB confirmed rats at 4 to 6 wk all showed SIB at 3 to 5 mo and at 12 to 13 mo after L-DOPA loading. To elucidate the brain locus important for the induction and cessation of SIB, in our study, we microinjected dopamine agonists and antagonists into various dopamine neuron innervating areas. L-DOPA-induced SIB was inhibited by the injection of a D1 antagonist, SCH-23390 (5 microg), into the bilateral substantia nigra, but not into the bilateral caudate-putamen or nucleus accumbens. The microinjection of YM-09151-2 (10 microg), a D2 antagonist, into these regions could not stop SIB. For examining the important area for the induction of SIB, we microinjected SKF-38393, D1 agonist, and/or LY-141865, D2 agonist (each 1 microg) into bilateral (or ipsilateral) caudate-putamen and substantia nigra. SIB was induced only in the case of D1 and D2 receptors in both the bilateral caudate putamen and bilateral substantia nigra being stimulated simultaneously by the mixed application of SKF-38393 and LY-141865. SIB was not induced by the sole injection of SKF-38393 into bilateral caudate-putamen or bilateral substantia nigra. These observations suggest that both caudate-putamen and nigral D1- and D2-like receptors are important for the induction of SIB, but, for cessation of SIB, up-regulated nigral D1 receptor is crucial.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗

PubMed1996

[Effect of microinjection of angiotensin II into area postrema on renal sodium excretion].

It has been known that area postrema (AP) possesses angiotensin II (A II) receptors and senses directly the change of blood A II level. The effect of microinjection of A II into the AP on renal sodium excretion was studied in the present investigation. Our experimental results show that microinjection of A II (2, 20 ng) increases GFR, RPF and UNaV (P < 0.05). These changes can be abolished by A II antagonist, saralatin and renal denervation (P < 0.05). However microinjection of A II (20 ng) into NTS produces no such changes. These results strongly suggest that the A II induced changes of renal hemodynamics and urine sodium excretion are mediated through AP.

Angiotensin II↗