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At least 253 records · Page 14Linked to original sources

PubMed1990

Neurotoxicity induced by the D-1 agonist SKF 38393 following microinjection into rat brain.

Microinjection of the D-1 agonist R-SKF 38393 into rat striatum resulted in extensive neurotoxic damage as revealed by the presence of large lesions upon histological analysis. These lesions were observed following a single injection of the drug into the ventrolateral striatum (30 micrograms/0.5 microliters) or nucleus accumbens (3.0 or 30 micrograms/0.5 microliters). This neurotoxic damage was also observed following microinjection of the inactive isomer S-SKF 38393. The mechanisms underlying these effects are presently unknown. The use of this compound for intracerebral microinjection studies should be re-evaluated. In view of the usefulness of central microinjections in studying the role of D-1 and D-2 receptors in behavior, it is suggested that alternative D-1 agonists, or ways of minimizing the toxicity of SKF 38393, should be considered.

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

PubMed1991

Anticonvulsant effects of ipsilateral but not contralateral microinjections of the dopamine D2 agonist LY 171555 into the nucleus accumbens of amygdala-kindled rats.

Recent radioligand binding studies demonstrated an increase in the density of dopamine D2 receptors in the nucleus accumbens ipsilateral to the stimulating electrode in amygdala- or hippocampal-kindled rats. In the present study we examined the anticonvulsant effect of dopamine agonists by unilateral microinjections into the nucleus accumbens in rats kindled from the right basolateral amygdaloid nucleus. Microinjections of the D2 agonist LY 171555 into the ipsilateral nucleus accumbens 15 min prior to the kindling stimulation in fully kindled rats decreased significantly kindling parameters such as seizure severity, seizure duration and afterdischarge duration, whereas the D1 agonist SKF 38393 had no anticonvulsant effects. After ipsilateral microinjection of 40 pmol LY 171555 focal and generalized kindled seizures were totally blocked in almost 50% of the rats. The anticonvulsant effect of LY 171555 could be completely antagonised by systemic administration of the D2 antagonist sulpiride. Microinjection of the D1 or D2 agonist into the nucleus accumbens contralateral to the stimulating electrode had no anticonvulsant effects. In accordance with other reports our data indicate a possible topographic limitation of D2 receptor mediated anticonvulsant effects to specific regions of the basal ganglia.

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

PubMed1992

Microinjection of L-glutamate into the nucleus tractus solitarii increases arterial pressure in conscious rats.

Microinjection of L-glutamate into the nucleus tractus solitarii (NTS) of anesthetized rats produces a fall in mean arterial pressure (MAP) similar to that observed during activation of baroreceptor afferents. In the present study we examined the effect of bilateral microinjections of L-glutamate through chronically implanted cannulae in the NTS of conscious freely moving rats. Group I (n = 6) was studied under conscious conditions and 24 h later the rats were anesthetized with urethane and the effects of L-glutamate re-examined. In conscious rats, L-glutamate (30 pmol to 5 nmol/100 nl) produced dose-dependent increases in MAP (+37 +/- 7 mmHg, 5 nmol), whereas under urethane anesthesia falls in MAP were observed (-11 +/- 3 mmHg, 5 nmol). Group II (n = 7) was studied under conscious conditions and 1 h later the rats were anesthetized with chloralose and the effects of L-glutamate re-examined. In this group of conscious rats L-glutamate (300 pmol to 5 nmol/100 nl) also produced dose-dependent increases in MAP (+37 +/- 5 mmHg, 5 nmol), whereas under chloralose anesthesia a dose-dependent depressor response was observed (-33 +/- 6 mmHg, 5 nmol). Saline microinjections into the NTS of conscious and anesthetized rats produced negligible effects. These data demonstrate that microinjection of L-glutamate into the NTS of rats produces a pressor response in conscious animals in contrast to depressor responses in animals anesthetized with chloralose or urethane.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia, General↗

PubMed1993

Microinjections of glutamate within trigeminal subnucleus interpolaris alters adrenal and autonomic function in the cat.

The influence of rostral portions of the trigeminal sensory complex on adrenal and autonomic function was assessed by microinjections of L-glutamate (500 or 5 mM, 100 nl) directed at subnucleus interpolaris (Vi) or at the nucleus principalis/subnucleus oralis level (Vp/Vo) in chloralose-anesthetized cats. Microinjections of glutamate (500 mM) within Vi evoked prompt (by +1 min) dose-related increases in the adrenal secretion of epinephrine (+11.4 +/- 2.5 ng/min, P < 0.001), adrenal blood flow (+0.19 +/- 0.06 ml/min, P < 0.05), mean arterial pressure (+6.6 +/- 3.0 mmHg, P < 0.025) and heart rate (+8.0 +/- 2.7 beats/min, P < 0.01, n = 16). Microinjections of lower doses of L-glutamate (5 mM, n = 7) within Vi had no effect. Microinjections of 500 mM glutamate within VP/Vo (n = 15) or within the spinal trigeminal tract (n = 13) had no consistent effect on adrenal or autonomic function. Plasma concentrations of ACTH were not altered significantly by glutamate regardless of dose or of the site of injection. The results suggest that local release of glutamate within Vi, but not within Vp/Vo, influences adrenal and autonomic function. Together with previous results obtained after injections of glutamate within subnucleus caudalis, these data indicate that glutaminergic input to both Vi and to more caudal portions of the spinal trigeminal nucleus contribute to the control of autonomic function such as that which often accompanies trigeminal nociception.

Adrenal Glands↗

PubMed1994

Cardiovascular changes elicited by microinjection of glycine or GABA into the spinal intermediolateral nucleus in urethane-anesthetized rats.

Sympathetic preganglionic neurons (SPNs), located in the intermediolateral cell column (IML) of the thoracolumbar spinal cord, contribute to the maintenance of arterial pressure (AP) and heart rate (HR) within normal limits under different physiological conditions. The purpose of this study was to examine the effects of microinjecting the putative inhibitory transmitters glycine (GLY) or gamma-amino-butyric acid (GABA) into functionally identified cardioacceleratory and vasoconstrictor IML sites at T1-T3. Rats were anesthetized (1.4 g/kg urethane i.p.), paralysed with decamethonium bromide (3.3 mg/kg i.v.) and artificially ventilated. Glutamate (GLU) microinjection (10-20 nl, pH = 7.4, 0.15 M in phosphate-buffered saline (PBS)) was used to identify 29 vasoconstrictor sites, of which 23 were also cardioacceleratory, in the right side of the T2 segment. GLY microinjection (10-20 nl, pH = 7.4, 0.5 M in PBS) into these sites resulted in significant decreases in both AP (n = 18) and HR (n = 14). The AP and HR responses to GLY were brief in duration and were attenuated by the specific GLY antagonist strychnine (STR). Microinjection of GABA (10-20 nl, pH = 7.4, 0.15-0.84 M in PBS, n = 6) and its agonist muscimol (10-20 nl, pH = 7.4, 0.9 mM in PBS, n = 6) into GLU-identified sites in the IML caused no changes in AP or HR. However, after the application of either GABA or muscimol, the changes in AP or HR elicited by GLU were eliminated, suggesting that GABA and muscimol decrease the excitability of SPNs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Microinjection of thyrotropin-releasing hormone in the paraventricular nucleus of the hypothalamus stimulates gastric contractility.

Changes in gastric contractility following microinjection of thyrotropin-releasing hormone (TRH) into the paraventricular nucleus of the hypothalamus (PVN) were examined in fasted, urethane-anesthetized rats. Gastric contractility was measured with extraluminal force transducers and analysed by computer. Unilateral and bilateral PVN microinjections of TRH (0.5 and 1.0 microgram) significantly increased the force index of gastric contractions from 0 to 60 min postinjection, when compared with animals microinjected with 0.1 microgram TRH, 0.1% BSA or TRH (0.5 and 1.0 microgram TRH) in sites adjacent to the PVN. The gastric force index was also significantly elevated from 61 to 120 min postinjection in rats receiving bilateral PVN microinjections of TRH (0.5 and 1.0 microgram). Peak gastric responses occurred within 10-20 min postinjection and represented an approximately eight-fold increase over basal values. In the remaining groups, the force index was not significantly altered from preinjection values. The excitatory action of TRH (1.0 microgram) on gastric contractility was completely abolished by subdiaphragmatic vagotomy. These results suggest that TRH acts within the PVN to stimulate gastric contractility via vagal-dependent pathways.

Animals↗

PubMed1988

Mechanism of Ca2+ release in medaka eggs microinjected with inositol 1,4,5-trisphosphate and Ca2+.

The reaction time of Ca2+ release from cytoplasmic stores induced by microinjection of inositol 1,4,5-trisphosphate (IP3), calcium ionophore A23187, Ca2+, Sr2+, Ba2+, and cyclic guanosine 5'-monophosphate (cGMP) in Oryzias latipes eggs in Ca2+-free medium was measured by the luminescence of aequorin injected into the egg. Microinjection of IP3 or calcium ionophore induced rapid Ca2+ release without a time lag, while microinjection of either Ca2+ or cGMP required a time lag of 5-30 sec for Ca2+ release. Following microinjection of both IP3 and Ca2+, Ca2+ release commenced in a cytoplasmic region close to the egg surface. These results suggest that in the medaka egg, cytoplasmic Ca2+ induces Ca2+ release from cytoplasmic stores indirectly, probably via a membrane factor such as IP3.

Aequorin↗

PubMed1992

Microinjection of S-nitrosocysteine into the nucleus tractus solitarii of conscious rats decreases arterial pressure but L-glutamate does not.

Unilateral microinjection of L-glutamate into the nucleus tractus solitarii of conscious rats increased arterial pressure and caused bradycardia while microinjection of S-nitrosocysteine into the same site of these animals caused hypotension and bradycardia. The responses to S-nitrosocysteine were blocked by prior microinjection of methylene blue into the nucleus tractus solitarii. The bradycardia and fall in arterial pressure induced by S-nitrosocysteine resemble more the cardiovascular changes in response to activation of baroreceptor afferents than the bradycardia and increase in arterial pressure induced by microinjection of L-glutamate into the nucleus tractus solitarii of conscious rats.

Animals↗

PubMed1984

Fate and structure of DNA microinjected into mouse TK-L cells.

Co-microinjection of single linearized molecules of plasmids containing the human beta-globin gene (pRK1) and the herpes simplex virus (HSV) type I thymidine kinase gene (pX1) into the mouse TK-L cell nucleus results in covalent linkage between these (or derived) molecules within the nucleus as revealed by Southern blotting, plasmid rescue, and recovery of plasmid-derived DNA from a Charon 4A phage library of cellular DNA. The microinjected DNA is predominantly found as high molecular weight DNA as determined by Hirt fractionation. Southern blotting data and recombinants from the Charon 4A library suggest that the plasmid DNA is in the form of a head-to-tail linear concatamer of up to 80 copies. Passage of these microinjected cells in selective medium (HAT) results in coordinate amplification of both plasmids, which are maintained in an approx. 3:1 molar ratio of pRK1 to pX1-derived molecules. Hybridization in situ shows the DNA to be integrated on a translocation chromosome, t(4;4). Integration does not appear to be site-specific, since plasmid DNA from another microinjected cell line, C2B, appears on a different translocation chromosome, t(8?;14). Plasmid rescue experiments confirm a previous finding that passage of pBR322 DNA through eukaryotic cells may result in deletions of normally stable plasmid DNA upon subsequent transformation of E. coli. These deletions appear to occur in the bacteria, and originate in a 128 bp region between the Sal I and Hae II sites of pBR322.

Animals↗

PubMed1986

Lysosomal uptake of isolated cell organelles microinjected into HeLa cells.

Lysosomes and microsomes were isolated from rat liver and microinjected into the cytoplasm of HeLa cells. The fate of the transplanted organelles and their effects on the recipient cells were followed in the electron microscope at various time intervals after administration. Needle injection with buffer or sucrose did not seem to evoke any ultrastructural alterations, such as induced autophagy or other signs of sublethal cell injury. Recipients of microinjected cell organelles elicited a rapid and conspicuous increase in membrane-bounded cytoplasmic vacuoles, concomitant with the disappearance of the injected material. Golgi complexes became abundant with many small vesicles clustering around their cisternae. The volume density of the lysosomal compartment increased 2-3-fold after organelle injection as compared with control-injected (0.3 M sucrose) or noninjected cells. Our preliminary results show that isolated cell organelles can be microinjected into cells n culture and indicate that the microinjected organelles were segregated from the cytoplasm into membrane-bounded vacuoles probably through autophagolysosome formation. Thus, this technique offers an additional approach for studies on the segregation and degradation of cell organelles in somatic cells and may enable more detailed analyses on the mechanisms of autophagic sequestration of specific cell organelles.

Animals↗

PubMed1992

Expression of microinjected DNA and RNA in early rabbit embryos: changes in permissiveness for expression and transcriptional selectivity.

Gene expression in rabbit early development was investigated by microinjecting LacZ DNA and LacZ RNA in 1-cell and 2-cell embryos. Expression of LacZ DNA could not be obtained before 30-36 hpf, although synthetic LacZ RNA was translated from 12 hpf at the least. The onset of expression of microinjected DNA correlated with the 8- to 16-cell stage. This suggests that before this stage, there is a general negative control of gene expression. The arrest of in vitro development at the 2- to 8-cell stages did not inhibit LacZ expression, which still occurred at 33 hpf. In addition the inhibition of the first cleavage by nocodazole resulted in LacZ expression in 1-cell embryos. Expression of microinjected DNA thus occurs at a fixed time after fertilization and is independent of cleavages and of the second and subsequent DNA replications. Therefore, the changes in permissiveness for the expression of microinjected DNA in rabbit embryos are reminiscent of those in mouse embryos. Transcriptional selectivity in rabbit embryos was compared to that in early mouse embryos. In both species, Sp1-sensitive promoters were active and the promoter of simian virus 40 did not require far upstream enhancers before late cleavage stages; genes driven by the -447, +563 region of murine leukemia virus were repressed. In rabbit, however, the H-2Kb promoter active in mouse was silent. Altogether, the results illustrate a remarkable conservation of the characteristics of the transcription in early rabbit and mouse embryos and the independence of its resumption from the pattern of cleavage.

Animals↗

PubMed1989

Microinjection of endogenous and exogenous proteins into primary cultures of rat hepatocytes and the degradation of the injected proteins.

1. A method to microinject proteins into cells through packaging proteins to erythrocyte ghosts (erythrocyte-mediated microinjection) was modified partially in order to apply the method to primary cultures of rat hepatocytes. 2. Degradation of the microinjected proteins was examined employing the improved method. The mean half-life of the injected endogenous liver protein was 20 hr. The data suggested that the injected proteins are degraded through both lysosomal and non-lysosomal proteolytic pathways probably depending on their structure. 3. The present method to microinject exogenous proteins into primary cultures of rat hepatocytes can be employed usefully for the investigations of protein metabolism in liver.

Animals↗

PubMed1996

Effects of microinjections of the neuropeptide substance P in the dorsal periaqueductal gray on the behaviour of rats in the plus-maze test.

Currently, the participation of neuropeptides in the generation of aversive states in the dorsal periaqueductal gray matter (DPAG) is poorly understood. The elevated plus maze (EPM) is widely used for studying the neurobiological mechanisms of anxiety in the laboratory. One difficulty with this test has been to evaluate the involvement of GABA mechanisms in the DPAG substrates of aversion, because microinjections of GABA receptor blockers in this region cause an intense behavioral activation. In this study, we examined in the EPM the effects of semicarbazide, a drug that acts indirectly on GABA neurotransmission through inhibition of the glutamic acid decarboxylase, and substance P (SP) following microinjections into the dorsal periaqueductal gray. Semicarbazide caused a clear decrease in the number of entries and time spent in the open arms. These results confirm previous data showing that GABA has a modulatory role in the DPAG, probably through reduction of tonic inhibitory mechanisms on neural substrates of aversion. A similar pattern of behavioral responses was observed with SP. However, these effects were more pronounced with intermediate doses of SP (25 ng), confirming the characteristic bell-shaped dose-effect function of this neuropeptide. The proaversive effects observed with DPAG microinjections of SP in the present study gain further relevance when combined with previous reports that have shown unconditioned and conditioned aversive effects following DPAG microinjections of SP in other animal models of aversion.

Animals↗

PubMed1983

Inhibition of SV40-induced cellular DNA synthesis by microinjection of monoclonal antibodies.

The region of the SV40 large T-antigen molecule recognized by a panel of monoclonal antibodies has been determined using hybrid Adeno-SV40 viruses, and manual microinjection of cloned deletion mutants. In addition, an investigation was made of how monoclonal antibodies microinjected into the nucleus can affect the ability of the T-antigen coding gene to stimulate cell DNA synthesis. The monoclonal antibody Pab 14, that recognized the -COOH terminal half of large T, was comicroinjected into quiescent cells together with plasmid pCl-1. This plasmid contains only that part of the T-antigen coding gene that extends from nucleotide residue 120, counterclockwise to nucleotide residue 4002, and makes a truncated T antigen 33,000 in molecular weight and missing the last 435 amino acids on the -COOH terminal side. Monoclonal antibody Pab 14 did not inhibit the stimulation of cellular DNA synthesis caused by microinjection of pCl-1, although it did inhibit cell DNA synthesis induced by microinjection of pSV2G, a recombinant plasmid that contains the entire T-antigen coding gene of SV40.

Adenoviruses, Human↗

PubMed1980

Reinforcing effects of morphine microinjection into the ventral tegmental area.

A neural substrate for the reinforcing property of an opiate drug was identified in the ventral tegmental area (VTA) by establishing conditioned reinforcement to salient environmental stimuli paired with intracerebral microinjections of morphine. Bilateral microinjections of morphine into the VTA in doses of 0.2 microgram and 1.0 microgram produced a subsequent change in place preference to a distinctive compartment previously associated with the stimulant effects of morphine. Microinjection of 1.0 microgram morphine at sites 2.5 mm dorsal to the VTA had no effect. Pretreatment with naloxone (2 mg/kg) antagonized the reinforcing effects of 1.0 microgram morphine as this group showed no significant change in place preference. Nor did control groups receiving microinjections of sterile physiological saline. Taken together, these data suggest that opiate receptors, located in the ventral tegmental area, play an important role in mediating the reinforcing effects of morphine. The possible involvement of dopaminergic neurons in these effects is discussed.

Animals↗

PubMed1986

Microinjections of methyl-beta-carboline-3-carboxylate into the dorsal raphe nucleus: behavioural consequences.

Very small quantities (0.01-10 ng) of the inverse benzodiazepine receptor agonist, methyl-beta-carboline-3-carboxylate (beta-CCM) microinjected into the dorsal raphe nucleus (DRN) of the rat selectively reduced social interaction, an effect consistent with an increase in anxiety. Similarly, intraperitoneally injected beta-CCM, within a limited dose range, reduced social interaction without affecting locomotor activity. The benzodiazepine receptor antagonist, RO15-1788 (1 ng) microinjected into the DRN, reversed the suppression of social interaction induced by either intra-raphe or intraperitoneal beta-CCM. Histological examination of the beta-CCM microinjection sites showed that locations within the DRN were almost invariably associated with decreases in social interaction; microinjections failing to decrease social interaction were located primarily outside the DRN. We conclude that the DRN has a major role in expressing the anxiogenic effect of beta-CCM and it may therefore be an important area in the neuronal system controlling anxiety.

Animals↗

PubMed1978

Phenotypic complementation of the SV40 tsA mutant defect in viral DNA synthesis following microinjection of SV40 T antigen.

African green monkey cells (CV-1P) were microinjected with highly purified SV40 T antigen using protein-loaded red cell ghosts and polyethylene glycol as fusagen. The microinjected cells were infected with a temperature-sensitive mutant of SV40 (tsA209) which is defective in the initiation of viral DNA synthesis. Using in situ hybridization as an assay method, we found that PEG-microinjection of both partially and highly purified T antigen resulted in an increase in the amount of viral DNA sequences in the monolayer. Moreover, 3H-thymidine-labeled and unlabeled Hirt supernatant from microinjected, tsA209-injected cells contained significantly more SV40 DNA than comparable extracts from sham-injected, tsA209-infected or uninfected cells, which were tested in parallel. Thus the introduction of highly purified, "large" SV40 T antigen led to phenotypic complementation of the tsA defect in viral DNA synthesis.

Antigens, Viral↗

PubMed1994

Cardiovascular effects of microinjection of atrial natriuretic factor (ANF) in the nucleus tractus solitarii of spontaneously hypertensive rats.

The effects on mean arterial pressure (MAP) and heart rate (HR) of unilateral microinjections of atrial natriuretic factor (ANF) into discrete sites of the nucleus tractus solitarii (NTS) of spontaneously hypertensive rats (SHR) were compared with those observed in normotensive Wistar-Kyoto rats (WKY). NTS sites were identified to be involved in cardiovascular control on the basis of the bradycardia and hypotension elicited by microinjections of 20 nl of 0.1 M L-glutamate. Microinjection of 20 nl of 10(-7) M ANF into 38 NTS 'cardiovascular sites' in rats of the SHR strain decreased MAP (-8.7 +/- 1.8 mmHg) and HR (-7.8 +/- 1.9 bpm) in 9 sites (24%), but caused no changes in the remaining 29 sites (76%). In WKY rats 35 cardiovascular sites within the NTS were studied. In 18 sites (51%) ANF microinjections induced a decrease in MAP (-15.1 +/- 1.9 mmHg) and in HR (-18.1 +/- 3.9 bpm), whereas the remaining 17 sites (49%) were unaffected. The decreased responsiveness of the NTS to ANF in the SHR animals could play a role in the development and/or maintenance of the elevated arterial blood pressure in genetically hypertensive rats.

Animals↗