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At least 1,189 records · Page 66Linked to original sources
PubMed1995

Effects of localized histamine microinjections into the hippocampal formation on the retrieval of a one-way active avoidance response in rats.

The possible role of histamine (HA) locally applied into the hippocampus on memory mechanisms of the rats was studied. The acquisition of a one-way active avoidance response to an ultrasonic 40 kHz sinus-wave tone anticipating an electric shock was used as experimental model. Learning sessions consisted in placing animals into a two compartment cage were they learnt to escape to the safe compartment after an ultrasonic tone anticipating an electric feet shock. After acquiring the conditioned avoidance response, animals were implanted with microinjection cannulae and injected with 1 microliter of saline, or increasing doses of histamine (9, 22.5, 45, and 90 nmol) into the hippocampus. In the experimental sessions, 4 trials before (PRE) and 4 trials afterward treatment (POST), the percentage of conditioned avoidance responses (% CAR) and the latency time to escape (LT) were measured. Results showed that HA increased significantly the LT and this effect was grossly dose-dependent. % CAR was also affected and the score was significantly inhibited by the imidazolamine administration. Results suggest that HA may be involved in memory retrieval processes in the hippocampus.

Acoustic Stimulation↗
PubMed1985

Effect of cell cycle position on transformation by microinjection.

We have investigated the effect of cell cycle position on the efficiency of transformation by microinjection. Linear recombinant plasmids transform synchronized cells with similar frequencies following injections at all cell cycle stages, whereas supercoiled molecules show a decreased ability to generate transformants in early S phase. This inhibition is not due to an inability to transiently express a transferred gene, since cells at all stages of the cycle efficiently expressed a hamster adenine phosphoribosyltransferase gene introduced on a supercoiled plasmid. Southern transfer analyses of the cell cycle specific transformants revealed that tandem arrays of plasmids, integrated into the host chromosomes, could be generated at all cell cycle stages.

Adenine Phosphoribosyltransferase↗
PubMed1996

FLP-mediated site-specific recombination in microinjected murine zygotes.

The FLP recombinase of yeast catalyses site-specific recombination between repeated FLP recombinase target (FRT) elements in yeast and in heterologous systems (Escherichia coli, Drosophila, mosquito and cultured mammalian cells). In this report, it is shown that transient FLP recombinase expression can recombine and activate an extrachromosomal silent reporter gene following coinjection into fertilized one-cell mouse eggs. Furthermore, it is demonstrated that introduction of a FLP-recombinase expression vector into transgenic one-cell fertilized mouse eggs induces a recombination event at a chromosomal FRT target locus. The resulting event occurred at the one-cell stage and deleted a chromosomal tandem array of a FRT containing lacZ expression cassette down to one or two copies. These results demonstrate that the FLP recombinase can be utilized to manipulate the genome of transgenic animals and suggest that FLP recombinase-mediated plasmid-to-chromosome targeting is feasible in microinjected eggs.

Animals↗
PubMed1991

Anxiolytic effect in the elevated plus-maze of the NMDA receptor antagonist AP7 microinjected into the dorsal periaqueductal grey.

In order to localise the often reported anxiolytic action of N-methyl-D-aspartate (NMDA) receptor antagonists, 2-amino-7-phosphonoheptanoic acid (AP7) was injected into the dorsal periaqueductal grey (DPAG) of rats exposed to the elevated plus-maze model of anxiety. Doses of 0.2, 2 and 20 nmol AP7 caused a dose-dependent increase in the percentage of open arm entries, the effect of the last two doses being significantly different from control. A non-significant tendency to increase the percentage of time spent on the open arms of the maze was also noticed. In contrast, the total number of entries into either the open or enclosed arms was not affected. Injections of AP7 localized outside the DPAG were ineffective. Therefore, microinjection of AP7 into the DPAG caused a selective anxiolytic effect in the elevated plus-maze. It may be suggested that the DPAG is a site of the anxiolytic action of NMDA antagonists reported following systemic administration.

2-Amino-5-phosphonovalerate↗
PubMed1991

Dopamine and conditioned reinforcement. I. Differential effects of amphetamine microinjections into striatal subregions.

In the conditioned reinforcement paradigm, animals learn a new instrumental response reinforced solely by conditioned reward (a stimulus that has previously been associated with primary reward). It has been shown that psychostimulants potentiate responding for conditioned reward and there is evidence that the nucleus accumbens is involved in this effect. The present experiments extend this work and examine the roles of various striatal subregions in the enhancement of responding for conditioned reward. In the conditioning phase, hungry rats were trained to associate a light/click stimulus with food delivery, with no levers present in the operant chamber. In the test phase, two levers were present and responding on one provided conditioned reward (presentation of the compound stimulus but no food). During this phase, microinjections of d-amphetamine (0, 0.2, 2.0, 20.0 micrograms/0.5 microliters) were made into seven striatal subregions in separate groups of rats. Injection of amphetamine into the nucleus accumbens elicited a dose-dependent, selective increase in responding for CR. Injections into posterior regions of the striatum had no effect. Significant and selective increases in CR responding were noted after injections into two regions neighboring the nucleus accumbens, the anterior dorsal and the ventromedial striatum, although the magnitude of these effects was considerably less than that following accumbens injections. Injections into ventrolateral regions increased responding in some rats, but this effect was very variable and not selective for the CR lever. These results are interpreted as evidence for functional heterogeneity of the striatum with regard to enhancement of conditioned reinforcement. The findings are discussed in relation to the theory that increased dopaminergic activity in the nucleus accumbens results in amplification of the response to a previously learned reward-related signal.

Animals↗
PubMed1993

Conditioned place preference induced by microinjection of 8-OH-DPAT into the dorsal or median raphe nucleus.

Experiments were conducted to examine the ability of the selective 5-hydroxytryptamine (5-HT)1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) to induce a conditioned place preference following peripheral injection, and direct microinjection into the dorsal or median raphe nuclei. An unbiased place preference paradigm was used in which control animals showed no preference for either of two compartments differing in terms of colour (white versus black), floor texture (rough versus smooth) and olfactory cues (no odour versus acetic acid odour). Drug treatments were paired with access to either of the two compartments, and saline injections were paired with access to the other compartment. Rats experiencing a low dose of 8-OH-DPAT (125 micrograms/kg) with a specific compartment demonstrated a significant preference for that compartment over one paired with saline injections. The magnitude of this effect was similar to that observed in rats treated with 1.5 mg/kg d-amphetamine. A significant place preference was found in animals receiving injections of 8-OH-DPAT in the dorsal raphe at 0.1 microgram but not 1 microgram. Animals also displayed a preference for the compartment paired with 1 microgram 8-OH-DPAT injected into the median raphe; lower doses were not effective. These results indicate that the mechanism by which 8-OH-DPAT induces a conditioned place preference involves activation of raphe somatodendritic 5-HT1A autoreceptors, leading to a reduction in 5-HT neurotransmission. This demonstration of the rewarding properties of 8-OH-DPAT, together with previous results showing increased feeding and sexual behaviour following 8-OH-DPAT treatment, strongly suggests an important role for brain 5-HT systems in reward and reinforcement processes.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1991

Microinjection of propranolol into the dorsal periaqueductal gray causes an anxiolytic effect in the elevated plus-maze antagonized by ritanserin.

The 5-HT1A/1B receptor antagonist propranolol was injected into the dorsal periaqueductal gray (DPAG) of rats exposed to the elevated plus-maze in order to investigate the participation in anxiety of 5-HT mechanisms operating in this brain region. Microinjection of D,L- or L-propranolol into the DPAG increased the percentage of total arm entries without affecting the total number of entries into either open or enclosed arms of the maze, an effect characteristic of anxiolytic drugs injected systemically. The doses of 5 nmol L-propranolol and 10 nmol D,L-propranolol caused anxiolytic effects of comparable magnitude, while the doses of 2.5 nmol of the former and 5 nmol of the latter were ineffective. Therefore, the L-isomer is likely to be the main one responsible for the pharmacological activity observed. In addition, the anxiolytic effect of 10 nmol D,L-propranolol was antagonized by 10 nmol of the 5-HT2/1C receptor antagonist ritanserin, previously injected into the DPAG. The present as well as previously reported results suggest that the anxiolytic effect of propranolol injected into the DPAG is due to increased release of 5-HT acting on post-synaptic 5-HT2 receptors, resultant from blockade of 5-HT1B autoreceptors that inhibit amine release from serotonergic nerve endings.

Analysis of Variance↗
PubMed1994

Differential behavioral effects following microinjection of an NMDA antagonist into nucleus accumbens subregions.

Recent studies have demonstrated the existence of two distinct regions within the nucleus accumbens (N.Acc) known as "core" and "shell". In order to investigate whether the behavioral functions of excitatory amino acid receptors differed between these two subregions, rats were administered microinjections of 2-amino-5-phosphonovaleric acid (AP-5), a competitive NMDA antagonist (0, 0.05, 0.2, 0.5, 1.0 microgram/0.5 microliter) into selected central and medial regions of the accumbens. The central and medial sites were assumed to correspond approximately to core and shell subregions, respectively. The animals were tested in two exploratory tasks: the open field and a novel object test. In the open field test, AP-5 significantly decreased peripheral locomotion and center rearing frequency in the central but not the medial group. Locomotion and rearing were not affected by AP5 infusion into a control site, the anterior dorsal striatum (ADS). In the novel object test, animals were tested in the same open field, with prior habituation, and with several novel objects placed within it. In this test, infusions of AP-5 (0, 1.0 microgram/0.5 microliter) decreased the number and duration of contacts with the novel objects in the central but not the medial group. In addition, peripheral and center locomotion were decreased by AP-5 infusions into the central site, whether objects were present or not. In contrast, AP-5 infusions into the medial site elicited an increase in peripheral locomotion in both stimulus conditions. These findings provide behavioral-pharmacological evidence that the central and medial subregions of the nucleus accumbens can be differentiated.(ABSTRACT TRUNCATED AT 250 WORDS)

2-Amino-5-phosphonovalerate↗
PubMed1990

GABAergic mechanisms in the cat red nucleus: effects of intracerebral microinjections of muscimol or bicuculline on a conditioned motor task.

Interneurons in the Red Nucleus (RN) are known to be under cortical control and to exert an inhibitory action, mediated by GABAergic mechanisms, on the main output towards the spinal cord. The effects of discrete injections of a GABA receptor agonist (muscimol) or an antagonist (bicuculline) in the Red Nucleus were tested on a motor task performed by seven cats. The subjects were trained to release a lever with a flexion movement of the forelimb controlled by a reaction time (RT) paradigm. Muscimol as well as bicuculline increased RTs in a dose-dependent manner at doses below 100 ng. However the parameters of the force exerted on the lever were differentially altered by the two drugs. Muscimol increased RTs by slowing down the force change preceding movement as well as slightly delaying its latency. While bicuculline increased drastically the force change latency. It could also speed up the force change velocity for low doses. At higher doses (up to 500 ng) both drugs produced an arrest of the performance either associated with anxiety signs (bicuculline) or dystonic movements of the head followed by body rotations (muscimol). The strong motor impairments as well as the disruption of the conditioned performances following muscimol or bicuculline microinjection in the RN suggest an important functional role for GABAergic interneurons. Under the control of cortical afferences they can modulate rubrospinal activity and participate in the triggering of a conditioned movement.

Animals↗
PubMed1996

Cytotoxic effects by microinjection of ADP-ribosylated skeletal muscle G-actin in PtK2 cells in the absence of Clostridium perfringens iota toxin.

The ADP-ribosylating toxins Clostridium botulinum C2 toxin and C. perfringens iota toxin, which ADP-ribosylate monomeric G-actin at Arg-177 but not the polymeric F-actin, induce depolymerization of the actin cytoskeleton in cultured cells. Since ADP-ribosylated G-actin has properties of a barbed-end-capping protein, we studied whether the ADP-ribosylated actin affects the actin cytoskeleton of PtK2 cells even in the absence of ADP-ribosylating toxin. Skeletal muscle actin was ADP-ribosylated by C. perfringens iota toxin and the toxin was removed using an anti-iota toxin antibody. Microinjection of ADP-ribosylated actin caused retraction of the cell body, redistribution and depolymerization of the actin cytoskeleton in a concentration- and time-dependent manner. The finding that ADP-ribosylated actin affects per se the actin cytoskeleton explains the cytopathic effects of ADP-ribosylating toxins on microfilaments, although F-actin is not directly modified by the toxins.

ADP Ribose Transferases↗
PubMedDate not supplied

The effects of atropine microinjections into the motor cortex of rats on the development of a motor habit.

Acetylcholine modulates the responses of motor cortex neurons during learning. Studies were carried out on the effects of atropine microinjections into the motor cortex on the development of a habit consisting of procuring food with the forelimbs in freely mobile rats. Animals received unilateral intracortical injections of atropine (5 and 15 micrograms in 0.6 microliter over 40 sec) in the forelimb representation area of the motor cortex. Rats were trained to procure food from a horizontal tube, using the forelimb contralateral to the injection site. Data were obtained which suggested that atropine has a dose-dependent effect on the development of this movement habit. The general effect of atropine doses consisted of suppressing the inhibition of concurrent movements during learning. The higher atropine concentration had a depressive effect on the initiation of the learned movement.

Animals↗
PubMed2004

Rewarding effects elicited by cocaine microinjections into the ventral tegmental area of C57BL/6 mice: involvement of dopamine D1 and serotonin1B receptors.

RATIONALE: The role of ventral tegmental area (VTA) in mediating the rewarding effects of cocaine has not been extensively studied. OBJECTIVES: We used the intracranial self-administration (ICSA) procedure to assess the involvement of the VTA in the rewarding effects of cocaine, and the effect of dopamine (DA) D(1)- and serotonin (5-HT)(1B)-receptor antagonists on ICSA of cocaine. METHODS: Adult male C57BL/6 mice were stereotaxically implanted, unilaterally, with a guide cannula either 1.5 or 2.3 mm above the VTA. After 1 week, mice were trained to discriminate between the two arms of a Y-maze over seven daily sessions, one arm being reinforced by intracranial cocaine microinjections. Starting from session 8, the D(1) and 5-HT(1B)-receptor antagonists were injected IP pre-test each day over five consecutive sessions. RESULTS: Mice injected into the VTA rapidly exhibited a preference for the cocaine-reinforced arm, whatever the dose of cocaine available (30 pmol or 150 pmol per injection), reaching optimum ICSA performance within 5 days. In contrast, mice injected 0.8 mm above the VTA did not discriminate between the arms of the maze and performed at random, except for one subject. Once the ICSA response was acquired, systemic pre-injections of either the D(1) (SCH23390; 25 microg/kg IP) or 5-HT(1B) (GR127935; 0.5 mg/kg IP) antagonist disrupted this behavior. Replacement of each antagonist by vehicle led to the reinstatement of intra-VTA cocaine self-administration. CONCLUSIONS: The results of the present study suggest that VTA neurons play a critical role in mediating the rewarding effects of acute cocaine and that both D(1) and 5-HT(1B) receptors modulate these effects.

Analysis of Variance↗
PubMed2005

Systemic and intra-accumbens microinjections of naltrexone interfere with tolerance to ethanol in rats.

RATIONALE: Evidence suggests a role for the opioid system in the control of ethanol reinforcement and drinking. Previous findings have shown that naltrexone, an opioid antagonist that decreases ethanol consumption in humans and experimental animals, reduces the acquisition of acute ethanol tolerance in rats. However, there are few data regarding the role of the opioid system in the acquisition of ethanol tolerance, particularly in brain areas involved in the rewarding actions of ethanol. OBJECTIVES: This study investigates the effects of systemic and of intra-accumbens injections of naltrexone on the development of rapid tolerance to ethanol. METHODS: Wistar rats received intraperitoneal injections of naltrexone (0.1-3.0 mg/kg) or microinjections into the core or shell portions of the nucleus accumbens (5-20 microg) before ethanol (2.7 g/kg i.p.). The animals were tested for motor coordination on the tilting plane apparatus. Tolerance was assessed 24 h later by administering the same dose of ethanol to all animals and retesting them on the tilting plane. RESULTS: The second injection of ethanol resulted in less motor incoordination on Day 2, suggesting the development of rapid tolerance. Pretreatment with naltrexone, either i.p. (0.3 and 0.6 mg/kg) or intra-accumbens (5-20 microg), on Day 1, blocked the development of rapid tolerance to the motor-incoordinating effects of ethanol on Day 2 without affecting the motor performance of the animals on Day 1. CONCLUSIONS: The results suggest that the opioid system may be involved in the development of ethanol tolerance, and that the nucleus accumbens may play a role in this phenomenon.

Animals↗
PubMed1996

Microinjections of phencyclidine (PCP) and related drugs into nucleus accumbens shell potentiate medial forebrain bundle brain stimulation reward.

Microinjections of phencyclidine (PCP) into the ventro-medial portion of nucleus accumbens in rats potentiated the rewarding impact of lateral hypothalamic brain stimulation. Similar effects were found with nomifensine, which shares with PCP the ability to block dopamine uptake and thus elevate synaptic dopamine levels but does not share with PCP the ability to block NMDA receptors. Similar effects were also seen with dizocilpine (MK-801) and [3-((+/-)2-carboxypiperazin-4-yl)propyl-1-phosphonate] (CPP), which share with PCP the ability to block NMDA receptors but not to block dopamine uptake. Thus PCP's properties as a dopamine uptake inhibitor and as an NMDA receptor antagonist each appear capable of producing reward-related actions in this brain region. The common denominator of these two PCP actions is decreased output of medium spiny neurons; these neurons are tonically activated by a glutamate projection from prefrontal cortex (PCP blocks this source of activation) and are tonically inhibited by a dopaminergic projection from the ventral tegmental area (PCP augments this inhibition).

Animals↗
PubMed1998

The role of the pedunculopontine tegmental nucleus in relation to conditioned motor performance in the cat. II. Effects of reversible inactivation by intracerebral microinjections.

The effects of reversible pharmacological manipulation of the neuronal activity in the pedunculopontine tegmental nucleus (PPTg) on the performance of a conditioned movement was studied in two freely moving cats. The microinjections were given in regions where, in the same subjects, we had previously identified neurons with context-dependent early activity after a trigger stimulus and with reinforcement-related activity. The subjects were conditioned to perform a forelimb-flexion movement controlled by a simple reaction-time task. In addition, one subject was trained to execute the same flexion movement, but delayed after the trigger stimulus. Food pellets were used as the reinforcer. Lidocaine injections (1 microl of 2% solution, injected over a 6-min period) induced a transient arrest of performance within minutes. The cessation of performance could be preceded by behavioral signs such as meowing, attempt to escape from the experimental booth, licking, or stereotyped posture. No rotational behavior could be observed. The effects of lidocaine could be mimicked in one subject by an extinction procedure. Muscimol injections (two injections of 0.2 microg in 1 microl, tested in one subject) also induced arrest of performance, but the return to pre-injection level of performance could not be obtained within the time of the test session. The quantitative analysis of reaction times and of inter-trial intervals showed that altering PPTg activity affected inter-trial intervals, but only slightly affected the reaction times. It is speculated that the PPTg is involved in the reinforcement process related to selecting the appropriate motor program.

Animals↗
PubMed1998

Microinjection of antibodies to centromere protein CENP-A arrests cells in interphase but does not prevent mitosis.

Centromere protein CENP-A is a histone H3-like protein associated specifically with the centromere and represents one of the human autoantigens identified by sera taken from patients with the CREST variant of progressive systemic sclerosis. Injection of whole human autoimmune serum to the centromere into interphase cells disrupts some mitotic events. It has been assumed that this effect is due to CENP-E and CENP-C autoantigens, because of the effects of injecting monospecific sera to those proteins into culture cells. Here we have used an antibody raised against an N-terminal peptide of the human autoantigen CENP-A to determine its function in mitosis and during cell cycle progression. Affinity-purified anti-CENP-A antibodies injected into the nucleus during the early replication stages of the cell cycle caused cells to arrest in interphase before mitosis. These cells showed highly condensed small nuclei, a granular cytoplasm and loss of their division capability. On the other hand, microinjection of nocodazole-blocked HeLa cells in mitosis resulted in the typical punctate staining pattern of CENP-A for centromeres during different stages of mitosis and apparently normal cell division. This was corroborated by time-lapse imaging microscopy analysis of mid-interphase-injected cells, revealing that they undergo mitosis and divide properly. However, a significant delay throughout the progression of mitotic stages was observed. These results suggest that CENP-A is involved predominantly in an essential interphase event at the centromere before mitosis. This may include chromatin assembly at the kinetochore coordinate with late replication of satellite DNA to form an active centromere.

Animals↗
PubMed2001

Microinjection reveals cell-to-cell movement of green fluorescent protein in cells of maize coleoptiles.

During the evaluation of dual-purpose plant/fungal expression systems, we found that green fluorescent protein (GFP) has the ability to move from cell to cell in the epidermis of Zea mays L. cv. Mutator coleoptiles as well as into underlying cortical cells. Movement of GFP was observed both when DNA encoding GFP and bacterially expressed GFP were microinjected into epidermal cells. This suggests that GFP is capable of cell-to-cell movement. From experiments using dextrans of known molecular weight linked to fluorescein isothiocyanate and tetramethylrhodamine isothiocyanate, we estimate that the plasmodesmata of these cells have a size exclusion limit < 4.4 kDa. Cell-to-cell GFP movement did not occur when GFP was altered to include a nucleus- or endoplasmic reticulum-retention sequence. The fact that these transcripts differ from that of cytoplasmic GFP by a small number of nucleotides suggests that the transcripts are not capable of movement, but movement of nucleic acid cannot be excluded. Since GFP is widely used to study cell-to-cell movement and to localize the expression of transgenes, caution should be exercised when interpreting results where GFP expression is used for localization.

Cell Communication↗