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At least 1,405 records · Page 78Linked to original sources

PubMed2002

Gastrin-releasing peptide microinjected into the amygdala inhibits feeding.

Bombesin (BN)-like peptides including gastrin-releasing peptide (GRP) are known to inhibit feeding. In the amygdaloid body BN receptors have been found in moderate to high densities. The central part of the amygdala (ACE) is essentially involved in the regulation of feeding and body weight. In the present experiments GRP was injected into the ACE and liquid food intake, general behavioural activity, as well as core temperature, were examined in male CFY rats. Food intake was measured every 5 min for 30 min and at the 40th and the 60th min following GRP or vehicle microinjections. Bilateral application of 50, 100 or 150 ng GRP resulted in transient inhibition of food intake while bilateral injection of 25 or 300 ng GRP did not modify feeding. Effect of GRP was eliminated by prior application of BN receptor antagonist [Leu(13)-psi(CH(2)NH)-Leu(14)]BN. After GRP or vehicle treatments animals were video-monitored and food intake, the first meal latency (FML), intermeal intervals (IMI), the time spent feeding (FT), grooming, resting and exploration were analysed at 5-min intervals for 30 min. However, FML did not change after GRP, the first IMI increased and intake, FT and intake/FT significantly decreased during the first 5 min. Duration of resting gradually increased after GRP and animals spent less time with exploration after GRP treatment than after vehicle injection. These differences were significant during the 25-30-min period. In body temperature, no significant changes were observed. Our results show that GRP in the ACE inhibits feeding and that GRP may decrease the efficiency of eating and may act as a satiety signal.

Amygdala↗

PubMed2003

Microinjection of renin-angiotensin system peptides in discrete sites within the rat periaqueductal gray matter elicits antinociception.

The intracerebroventricular administration of renin substrate or angiotensin II evokes antinociception in rodents, but the brain sites where most of the renin-angiotensin system peptides act are not yet known. This study describes the antinociceptive effects of microinjecting porcine renin substrate tetradecapeptide (RS) or angiotensins I (AI), II (AII) or III (AIII) into different regions of the periaqueductal gray matter (PAG), using the rat tail flick test. All the above peptides were effective following administration into several PAG regions. Their antinociceptive effects were strongly evoked from the caudal ventrolateral and ventral PAG, including the dorsal raphe nucleus. A dose-dependent antinociception following administration into the ventrolateral PAG was demonstrated for all peptides studied. The effect of AII from the ventrolateral PAG was inhibited by the previous local administration of saralasin, a non-selective angiotensin receptor antagonist. Moreover, the peak effects of RS and AI occurred later than those of AII and AIII. The time-course of antinociception suggests that longer-chain peptides are locally processed to biologically active smaller-chain peptides. This study shows for the first time the antinociceptive effect of RS, AI, AII and III in well-defined PAG regions, an effect that is receptor mediated for AII.

Angiotensinogen↗

PubMed2003

Angiotensin II and III microinjections into the zona incerta influence drinking behavior.

Different doses of angiotensin II (AII) or angiotensin III (AIII) microinjections into the zona incerta have been studied on drinking of rats in separate experiments during the consequent 60-min-daily-drinking period. Also, the dipsogen power of only the effective dose of AII and AIII was compared to vehicle treated rats. After, angiotensin receptor (AT(1), AT(2)) antagonists on AII or AIII induced drinking were tested. In the first and second experiments only the 100 ng AII and the 200 ng AIII increased water intake significantly. In the third experiment the AII started its dipsogen effect earlier, at the 5 min measuring time, compared to the AIII. Both effects kept on lasting parallel from the 10 min on. Considering the antagonist pre-treatments in the fourth experiment, animals were injected with 90 ng losartan, an AT(1) antagonist, 180 ng PD 123319 or 200 ng CGP 42112, both AT(2) antagonists, respectively. Both AII and AIII increased water consumption. The effect of AII could be blocked by losartan, but not by PD 123319 or CGP 42112. On the other hand, the effect of AIII could not be blocked by losartan, but by both the PD 123319 and CGP 42112. Since, the effects of AII, AIII and angiotensin antagonists have not been tested in the zona incerta, the finding that water intake increased after AII or AIII injections and it could be blocked only by either of the antagonists suggests that AT(1) and AT(2) receptors play partially different roles in the regulation of water intake.

Angiotensin II↗

PubMed1997

Microinjection of NMDA into the SCN region mimics the phase shifting effect of light in hamsters.

Although there is considerable data that glutamate is the primary transducer of photic information to the circadian clock in the suprachiasmatic nucleus (SCN), the ability of glutamate to mimic the phase-shifting effects of light has yet to be demonstrated in vivo. In the present study, microinjections of the glutamate agonist NMDA directly into the SCN of Syrian hamsters induced significant phase delays at circadian time (CT) 13.5 and phase advances at CT 19. These results support the hypothesis that glutamate is the primary neurotransmitter responsible for the transduction of photic information to the SCN.

Animals↗

PubMed1997

PAG-microinjected dipyrone (metamizol) inhibits responses of spinal dorsal horn neurons to natural noxious stimulation in rats.

In addition to their well-known peripheral and spinal effects, non-steroidal antiinflammatory drugs (NSAIDs) are believed to diminish nociceptive responses by acting supraspinally and activating descending modulatory systems. We have herein investigated whether this descending action involves a depression of spinal sensory neurons. In rats under barbiturate anesthesia, responses of lumbar wide-dynamic-range neurons to a noxious clamp in their receptive fields were depressed to 46% of baseline value by the microinjection of 100 microg dipyrone (metamizol) into the periaqueductal gray matter (PAG). These results show that PAG application of NSAIDs activates descending systems which depress the excitation of spinal sensory neurons by natural noxious stimuli.

Animals↗

PubMed1997

Mapping of locomotor behavioral arousal induced by microinjections of dopamine within nucleus accumbens septi of rat forebrain.

Dopamine (DA) at ca. ED50 (16 microg) or saline was stereotaxically microinjected unilaterally 2 h after pretreatment with an MAO inhibitor into left or right nucleus accumbens septi of 697 freely moving rats (1394 injections) to define subregions involved in DA-induced behavioral arousal throughout the anatomical extent of the accumbens. Locomotion was quantified electronically and behavioral responses were assigned to histologically verified injection sites; postural or stereotyped behaviors characteristic of DA injections in caudate-putamen did not occur. Screening with 60 injections across mid-accumbens (2.2-3.2 mm rostral to bregma) indicated that locomotion was elicited non-homogeneously, and was particularly intense dorsomedially. Sites yielding intense arousal and their inactive surround were mapped along the rostrocaudal axis (1.4-4.2 mm anterior to bregma) in coronal sections. Responses to DA showed lateral symmetry and were similar across rostrocaudal levels, with intense responses in dorsomedial accumbens along its border with the caudate-putamen. This functional localization does not coincide closely with reported distributions of DA or its receptors, nor with histologically or histochemically defined core-shell regions of this limbic structure. Nucleus accumbens in rat brain thus appears to be organized functionally into distinct subregions differing markedly in ability to produce locomotor hyperactivity in response to exogenous DA.

Animals↗

PubMed1998

Ipsilateral turning behavior induced by unilateral microinjections of a cannabinoid into the rat subthalamic nucleus.

The subthalamic nucleus contains cannabinoid receptors and cannabinoid receptor mRNA. However, the role of cannabinoid receptors in this nucleus has not been examined. In order to investigate the functional role of cannabinoid receptors in the rat subthalamic nucleus, turning activity was observed following unilateral microinjection of the synthetic cannabinoid CP 55,940. CP 55,940 (10 microg) induced ipsilateral turning. This effect was blocked by coadministration of the cannabinoid receptor antagonist SR141716A (5 microg). These results suggest that cannabinoid receptors in the subthalamic nucleus mediate an inhibition of motor activity.

Animals↗

PubMed1998

Microinjections of anisomycin into the intermediate cerebellum during learning affect the acquisition of classically conditioned responses in the rabbit.

The purpose of this study was to examine the effects of protein synthesis inhibition in the intermediate cerebellum on the acquisition and expression of classically conditioned nictitating membrane responses in the rabbit. Animals were conditioned for three days in a standard delay paradigm. Before each training session, either a solution of anisomycin (a protein synthesis inhibitor) or vehicle was bilaterally injected into the interposed cerebellar nuclear. Following these three training sessions, rabbits were tested to determine whether the previous training under the influence of anisomycin or vehicle resulted in the acquisition of conditioned responses. In this test, animals that were injected previously with the protein synthesis inhibitor exhibited significantly less retention of conditioned responses than rabbits injected with vehicle. Additional experiments demonstrated that anisomycin does not block the expression of conditioned responses during conditioning or in well-trained animals. Microinjections of muscimol at the same sites of the previous drug infusions suppressed the expression of conditioned responses, indicating that the protein synthesis inhibitor was applied to the eyeblink-related parts of cerebellar circuits. The obtained data are the first to demonstrate that a manipulation of cerebellar circuits, which does not affect the performance of learned behavior, can affect the process of learning. These results suggest that the synthesis of new proteins in the intermediate cerebellum participates in the formation of plastic changes responsible for eyeblink conditioning.

Analysis of Variance↗

PubMed1998

Noxious heat-evoked fos-like immunoreactivity in the rat lumbar dorsal horn is inhibited by glutamate microinjections in the upper cervical spinal cord.

Microinjections of glutamate into the upper cervical spinal cord significantly reduced (to 57% of control) the total number of neurons demonstrating noxious heat-evoked fos-like immunoreactivity in the lumbar spinal cord. Neurons in the upper cervical spinal cord, with descending propriospinal projections to the lumbar spinal cord, therefore, produce inhibitory effects on dorsal horn neurons in the lumbar spinal cord that receive nociceptive input from cutaneous thermal nociceptors.

Animals↗

PubMed1998

Microinjection of morphine into the rostral ventromedial medulla produces greater antinociception in male compared to female rats.

The antinociceptive and locomotor effects of microinjecting morphine into the rostral ventromedial medulla (RVM) of male and female rats was assessed. Male rats showed greater antinociception than female rats at all doses and times following morphine administration. Male, but not female rats, also showed a dose dependent decrease in locomotion. These data demonstrate that sex differences in antinociception are mediated at least in part by the RVM.

Animals↗

PubMed1999

CGRP microinjection into the ventromedial or dorsomedial hypothalamic nucleus activates heat production.

Unilateral microinjection of calcitonin gene-related peptide (CGRP, 1.6 pmol; 0.2 microl) into the ventromedial hypothalamus (VMH) and dorsomedial hypothalamus (DMH) immediately increased oxygen consumption (VO2), heart rate (HR), colonic temperature (Tco), and temperature of interscapular brown adipose tissue (TIBAT) in urethane-anesthetized rats, whereas vehicle saline injection into the VMH and CGRP injection into other hypothalamic regions such as the preoptic area, lateral hypothalamic area, paraventricular nucleus, and bed nucleus of the stria terminalis had no effect. The effects of CGRP injection into the VMH were dose-dependent over the range of 0.016-1.6 pmol. CGRP administration to the lateral ventricle (LV) required 16-320 pmol to elicit similar degrees of responses that were observed after the injection into the VMH. The increase in TIBAT was always higher than that in Tco after CGRP injection. Injection of [Cys(ACM)2,7]hCGRPalpha, a selective CGRP2 receptor agonist, did not induce any thermogenic effects. Human CGRP8-37, a proposed CGRP1 receptor antagonist, by itself induced heat production responses with no signs of inhibition of CGRP-induced responses. Thus, the receptor subtype of the thermogenic effect of CGRP could not be determined by the available pharmacological tools. The present results show that centrally administrated CGRP induces heat production in the BAT specifically through the VMH or DMH.

Adipose Tissue, Brown↗

PubMed1999

Anxiolytic effects of the neuroactive steroid pregnanolone (3 alpha-OH-5 beta-pregnan-20-one) after microinjection in the dorsal hippocampus and lateral septum.

The anxiolytic effects of the neuroactive steroid, 3 alpha-OH-5 beta-pregnan-20-one (pregnanolone), were determined after injection into the dorsal hippocampus or lateral septum in adult male rats. An increase in the proportion of time spent on the open arms of the elevated plus-maze was found after 2.5 and 5 micrograms of pregnanolone in the hippocampus, but not in the lateral septum. Intrahippocampal injection of 2.5 micrograms of the 3 beta-epimer of pregnanolone did not affect behavior in the plus-maze; a higher dose of 5 micrograms produced an anxiogenic effect. In the shock-probe burying test latency to burying behavior was increased by intrahippocampal or intraseptal injection of 2.5 and 5 micrograms of pregnanolone; the duration of burying behavior was decreased by 0.5, 2.5 and 5 micrograms of pregnanolone injection in the dorsal hippocampus or lateral septum. The number of contacts with the shock probe was not affected by any dose of pregnanolone in either intracranial site of injection. The anxiolytic effects of intrahippocampal or intraseptal injection of pregnanolone were blocked by intracranial pretreatment with 20 ng of picrotoxin, but not by microinjection of 5 micrograms of flumazenil or 200 ng of PK 11195. Thus, inhibition of the hippocampus, mediated by the pregnanolone's action at the GABAA receptor, produces a general anxiolytic effect. However, similar inhibition in the lateral septum attenuates active avoidance of anxiogenic stimuli (i.e., decreased burying behavior), but not passive avoidance of aversive stimuli (i.e., exploration of open arms of the plus-maze and number of shocks in the probe burying test).

Animals↗

PubMed2000

Sleep-inducing effects of adenosine microinjections into the medial preoptic area are blocked by flumazenil.

Microinjection of a wide range of sedative agents, including triazolam, pentobarbital, ethanol and adenosine, into the medial preoptic area has been shown to increase sleep, suggesting that it is an important (though not necessarily the only) anatomic site mediating hypnotic effects of these compounds. The mechanism by which adenosine increases sleep at this site is not clear, but one possibility is that this is related to its effects on the GABA(A)-benzodiazepine receptor complex. In order to assess this possibility, this paper describes the administration of adenosine, alone and in combination with the benzodiazepine receptor blocker flumazenil, into the MPA. It was found that 12.5 and 25 nM of adenosine significantly reduced sleep latency and increased total sleep time. The sleep-inducing effect was blocked by flumazenil. Flumazenil caused a modest increase in total sleep, and prevented the increase in total sleep induced by the higher dose of adenosine. These data suggest that at least one aspect of the hypnotic properties of adenosine is mediated by a direct or indirect action on the GABA(A)-benzodiazepine receptor complex.

Adenosine↗

PubMed2003

Diffusion-limited compartmentalization of mammalian cell nuclei assessed by microinjected macromolecules.

In order to investigate the accessibility of the nucleoplasm for macromolecules with different physical properties, we microinjected FITC-conjugated dextrans of different sizes as well as anionic FITC-dextrans and FITC-poly-L-lysine into mammalian cell nuclei. Small dextrans displayed a homogeneous nuclear distribution. With increasing molecular mass (42 to 2500 kDa), FITC-dextrans were progressively excluded from chromatin regions, accumulating in and thereby outlining an apparently extended interchromatin space. Anionic FITC-dextrans (500 kDa) showed complete exclusion from labeled chromatin regions, while the positively charged FITC-poly-L-lysine was to some extent present within the chromatin regions. Moreover, the FITC-poly-L-lysine preferentially localized at the nuclear periphery. We also found a size-dependent exclusion of FITC-dextrans from nucleoli regions, while the FITC-poly-L-lysine accumulated in the nucleoli. Thus, the distinct and restricted nuclear accessibility for macromolecules is dependent on molecule size and electrical charge.

Animals↗

PubMed2000

Priming of PC12 cells for semiquantitative microinjection studies involving Ras.

Nerve growth factor and activated Ras can induce differentiation of rat pheochromocytoma cells (PC12 cells) [Greene and Tischler (1976) Proc. Natl. Acad. Sci. USA 73, 2424-2428] from a chromaffin cell-like morphology into one that resembles sympathetic neurones. We developed a special treatment of PC12 cells which apparently synchronizes these cells such that they are more useful for semi-quantitative microinjection studies for signal transduction pathways. This treatment leads to a faster and more reproducible differentiation which faithfully reproduces the involvement of Ras in the process and allows a comparison of the biological activity of different Ras mutants. It shows that G12V and Q61L oncogenic mutants are not equally potent in inducing differentiation. Partial loss-of-function mutations T35S, E37G and Y40C are inactive and even a triple combination of these does not restore full biological activity.

Amino Acid Substitution↗

PubMed2000

Recombinant C1b domain of PKCdelta triggers meiotic maturation upon microinjection in Xenopus laevis oocytes.

The C1 domains are 50 amino acid sequences present in protein kinase C (PKC) isozymes that are responsible for binding of phorbol esters and the lipid second messenger diacylglycerol (DAG). We found that bacterially expressed C1b domain of PKCdelta induces germinal vesicle breakdown (GVBD) when microinjected into Xenopus laevis oocytes. Injection of the C1b domain of PKCdelta significantly enhanced insulin- but not progesterone-induced maturation. Interestingly, the PKCdelta C1b domain markedly synergized with normal Ras protein to induce oocyte maturation when both proteins were co-injected in oocytes. Our results demonstrate that the purified C1b domain of PKCdelta is sufficient to promote meiotic maturation of X. laevis oocytes probably through activation of components of the insulin/Ras signaling pathway.

Amino Acid Substitution↗

PubMed1998

Recombinant protein kinase C-gamma phorbol binding domain upon microinjection blocked insulin-induced maturation of Xenopus laevis oocytes.

The second cysteine-rich (Cys-2) domain of rat brain PKC-gamma regulatory region C1 (92-173) was expressed in Escherichia coli cells and purified. NMR studies of Cys-2 protein identified the phorbol and other phospholipid binding sites within this molecule (Xu, R.X., Pawelczyk, T., Xia, T-H. and Brown, S.T. (1997) Biochemistry 37, 10709-10717). Here, we tested the ability of this domain to bind other proteins. Using an overlay assay we show that the Cys-2 domain binds other proteins in Xenopus oocyte soluble fraction. Unlike the kinase activity, binding of Cys-2 to other proteins was detected in the absence of added phospholipids. Microinjection of Cys-2 protein into Xenopus leavis oocytes inhibited insulin-induced but not progesterone-induced maturation. The smallest dose that enhanced insulin-induced maturation was 0.45 x 10(-12) mol injected Cys-2. These results demonstrate that the PKC-gamma Cys-2 domain beside being the binding site for phorbol ester/DAG and phosphatidylserine binds also other proteins. The proteins that interact with Cys-2 domain of PKC are essential for insulin-induced maturation program in oocytes.

Animals↗

PubMed1999

Microinjected glutathione reductase crystals as indicators of the redox status in living cells.

The flavoenzyme glutathione reductase catalyses electron transfer reactions between two major intracellular redox buffers, namely the NADPH/NADP+ couple and the 2 glutathione/glutathione disulfide couple. On this account, microcrystals of the enzyme were tested as redox probes of intracellular compartments. For introducing protein crystals into human fibroblasts, different methods (microinjection, particle bombardment and optical tweezers) were explored and compared. When glutathione reductase crystals are present in a cytosolic environment, the transition of the yellow Eox form to the orange-red 2-electron reduced charge transfer form, EH2, is observed. Taking into account the midpoint potential of the Eox/EH2 couple, the redox potential of the cytosol was found to be < -270 mV at pH 7.4 and 37 degrees C. As a general conclusion, competent proteins in crystalline--that is signal-amplifying--form are promising probes for studying intracellular events.

Cells, Cultured↗