Repeated microinjections into the medial prefrontal cortex (mPFC) impair extinction of conditioned place preference in mice — TEI-tagged view

Groblewski, Cunningham · 2012 · neuroscience · doi:10.1016/j.bbr.2012.01.059 · full text at the source · TEI xml · tagged edition · index · concepts · authors
Where this record's text came from. faceted 2026-08 by reading the locally held text (xml); every evidence string verified as an exact substring of that text before merging. Claim type is 'null-result' for the intended comparison (bupivacaine did not differ from vehicle; neither group extinguished), but the paper is equally a methodological critique: its closing claim is that the microinjection procedure as performed is unsuitable for this paradigm, which is recorded under limitations rather than duplicated as the claim shape. Replication status is 'untested' in the sense of independent replication; the paper does report internal reproduction, stating that 'the results of Experiment 2 are similar to those in Experiment 1' and that an extended replicate of Experiment 1 with six days of extinction gave the same persistent CPP. Roughly a quarter of subjects were excluded a priori as non-learners or on histological criteria, which the paper states but does not frame as a limitation of its own conclusions. 2026-08-12 and 2026-08-15 schema-0.3 passes: every quote verified as an exact substring of the staged text and anchored at merge time; concept terms verified present; every edge endpoint an existing node.
6 quotations on this page contain a reference number where the paper printed a superscript. This journal style prints citations as raised numerals — parameters12,13 — and the extraction flattened them into the line, so the sentence reads parameters 12 , 13 . The digits are citation markers and not part of what the author wrote. The quotes keep them for the same reason they keep every other extraction fault: a quote is reproduced exactly or it proves nothing.
Tags on this paper every tag the record carries, whether or not the words occur in the text below — a number is how many papers in the corpus carry that tag; a tag with no number is unique to this paper
medial prefrontal cortex4 conditioned place preference extinction3 bupivacaine microinjection vehicle3 No Sx prelimbic3 infralimbic2 guide cannula lesion2 nucleus accumbens basolateral amygdala complex conditioned stimuli histological criteria conditioned fear behavioural-experiment117

Our tag layer for this paper

Cross-disciplinary analysis
content-tags & facets: phenomenon · claim-type · cross-domain-transfer · mechanism · claim-relation · replication-status · limitation — grounded in the full text
phenomenonextinction of ethanol-induced conditioned place preference— “The medial prefrontal cortex (mPFC) is important for extinction of many behaviors including conditioned place preference (CPP).”
claim-typenull-result— “Thus, neither the control (Vehicle) nor the experimental group (Bupivacaine) showed significant extinction of EtOH-CPP.”
method — evidence— “We examined the effects of intra-mPFC inactivation (with bupivacaine) on extinction of ethanol-induced CPP in mice.”
object of study — evidence— “We examined the effects of intra-mPFC inactivation (with bupivacaine) on extinction of ethanol-induced CPP in mice.”
mechanismrepeated cannula and injector insertion destroys tissue in Layers I-III of the mPFC on both sides of the midline, removing projection neurons that normally regulate the nucleus accumbens and basolateral amygdala, and the loss of that top-down regulation is what blocks extinction learning— “Therefore, it is conceivable that the damage caused by the microinjection procedure resulted in altered mPFC regulation of the NAc and BLC, thereby impairing extinction of EtOH-CPP.”mechanisms[0]
mechanismcumulative mechanical damage from implantation plus repeated infusions amounts to a partial lesion of a region already shown to be necessary for extinction, so the injection itself reproduces the lesion effect regardless of what is infused— “The most parsimonious explanation for these findings is that the microinjection procedure, regardless of the compound being injected, caused tissue damage to an area of the brain shown to be necessary for extinction of EtOH-CPP [ 1 ].”mechanisms[1]
mechanismThe vehicle group could not serve as the control, because whatever impaired extinction affected vehicle and drug alike— “Because of the incomplete extinction in the Vehicle group, it remained unknown if inactivation of the mPFC with bupivacaine was able to impair extinction.”mechanisms[2]
mechanismOnly a group that never had the procedure could reveal it, which is why the no-surgery control was added in the second experiment— “Finally, an additional control group that did not receive surgery was included (No Sx group) in order to assess the effects of the handling portion of the microinjection procedure on extinction learning.”mechanisms[3]
mechanismThe proposed cause: the repeated procedure damages the very region it is delivering into, amounting to a partial lesion— “The most parsimonious explanation for these findings is that the microinjection procedure, regardless of the compound being injected, caused tissue damage to an area of the brain shown to be necessary for extinction of EtOH-CPP [ 1 ].”mechanisms[4]
mechanismBecause the cannula sat on the midline, the damage was bilateral rather than confined to one hemisphere— “Importantly, because these cannulae were located on the sagital midline, tissue damage to these layers of the cortex affected both hemispheres.”mechanisms[5]
mechanismThe damaged layers contain projection neurons to accumbens and amygdala whose activity changes with drug cues, which is a route from local damage to the behavioural failure— “Retrograde tracing experiments have shown that Layers II/III of the mPFC contain dense populations of projection neurons that terminate in both the nucleus accumbens (NAc) and basolateral amygdala complex…mechanisms[6]
mechanismSurgery and anaesthesia were ruled out by comparison with their own earlier study matched on strain, frame, coordinates, anaesthesia and surgeonmechanisms[7]
extends Groblewski et al. mPFC lesion findings [1] and the rat bupivacaine microinjection procedure [2]
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claim_relations[0]
builds-on Groblewski et al. (2011) [1] electrode-insertion control result
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claim_relations[1]
extends Groblewski et al. on mPFC lesions impairing extinction of ethanol place preference
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claim_relations[2]
critiques the repeated intra-mPFC microinjection procedure as a tool for multi-day extinction studies
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claim_relations[3]
sensecontrol - VEHICLE: everything but the drug, which is the standard control and here shares the confound
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senses[0]
sensecontrol - NO PROCEDURE AT ALL: no surgery, no cannula, no injection, and the only group in which extinction occurred
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senses[1]
sensethe manipulation - as intended: reversible pharmacological inactivation of a region during extinction trials
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senses[2]
sensethe manipulation - as delivered: cumulative mechanical damage from repeating the delivery, which is a lesion and not an inactivation
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senses[3]
senseextinction procedure - single-session, as in conditioned fear, where one injection suffices and this problem never arises
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senses[4]
senseextinction procedure - multi-day, as required for ethanol place preference, where the injections must be repeated and the damage accumulates
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senses[5]
controlan additional group given NO SURGERY AT ALL (the No Sx group), alongside the vehicle-infused and drug-infused groups — held — rules out the handling involved in the microinjection procedure - being restrained and manipulated - as the cause of the change in extinction learning, which a vehicle control alone cannot separate because it is handled too
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controls[0]
differs inthe repeated intra-mPFC microinjection procedure as a tool for multi-day extinction studies — THE AXIS IS THE NUMBER OF INJECTIONS, AND MINIMISING IT DOES NOT HELP. 'the damage caused by intra-mPFC injections even when the number of injections was minimized, was sufficient enough to impair extinction - an effect that persisted despite extended extinction training.' The critique of the procedure is dosed: it survives the obvious remedy.
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claim_relations[3]
auditmicroinjection —causes→ lesion — Audit 2026-09-28: DOUBTFUL - read against its full span and kept, but the tissue damage 'MAY HAVE BEEN sufficient' to amount to a lesion; causes is hedged.
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concept_edges[0]
weightmicroinjection —causes→ lesion — 0.9
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concept_edges[0]
edgemicroinjection —causes→ extinction
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concept_edges[1]
edgemedial prefrontal cortex —causes→ extinction
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concept_edges[2]
retractedEVIDENCE CORRECTED (RE-GROUNDED) · vehicle —contrasted-with→ No Sx — RE-GROUNDED. The old span named only the No Sx group. This sentence names both and carries the contrast with its direction - 'the No Sx, BUT NOT Vehicle or Bupivacaine, group…'.
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concept_edges[3]
edgeprelimbic —part-of→ medial prefrontal cortex
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concept_edges[5]
edgeinfralimbic —part-of→ medial prefrontal cortex
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concept_edges[6]
auditguide cannula —part-of→ microinjection — Audit 2026-09-28: DOUBTFUL - read against its full span and kept, but a guide cannula was IMPLANTED for the microinjections; part-of is loose.
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concept_edges[7]
auditconditioned place preference —operationalised-as→ conditioned stimuli — Audit 2026-09-28: DOUBTFUL - read against its full span and kept, but the conditioned stimuli were the tactile cues of the place-preference task; operationalised-as is loose.
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concept_edges[9]
differs inconditioned fear —contrasted-with→ conditioned place preference — THE AXIS IS HOW MUCH TRAINING EXTINCTION TAKES. 'Unlike extinction of conditioned fear, however, extinction of EtOH-CPP requires multiple extinction trials over the course of several days.' Both are extinction; they differ on the schedule needed, which is why a procedure validated on one can fail on the other for reasons that have nothing to do with the mechanism.
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concept_edges[10]
outcome (stated by the tagger, no quotation)control — heldcontrols[0]
for which uselimitation — every within-subject comparison reported: THIS METHOD OF ANALYSIS, ALTHOUGH NOT STATISTICALLY JUSTIFIED, WAS IMPLEMENTED BECAUSE OF THE NARROW DETECTION WINDOW AND INCREASED VARIABILITY inherent to the paradigm
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limitations[0]
for which uselimitation — the projection-damage explanation: FURTHER EXAMINATION OF THE EFFECTS OF THE MICROINJECTION PROCEDURE ON THESE SPECIFIC PROJECTIONS ... IS REQUIRED - proposed, not tested
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limitations[1]
for which uselimitation — any pharmacological use of this procedure: IRRESPECTIVE OF HOW EACH COMPONENT ... CONTRIBUTES TO THE OBSERVED EFFECT, AS CURRENTLY PERFORMED, THIS PROCEDURE IS NOT ADEQUATELY SUITED TO EXAMINE THE EFFECTS OF INTRA-mPFC PHARMACOLOGICAL MANIPULATIONS ON EXTINCTION
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limitations[2]
duplicate oflimitation — limitations[0]
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limitations[3]
for which uselimitation — the same scope as limitations[0] - the within-subject comparisons are acknowledged as not statistically justified, with the reason given
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limitations[3]
for which uselimitation — the sample behind Experiment 1: A TOTAL OF 18 ANIMALS WERE REMOVED FROM ANALYSIS (9 SHOWED LESS THAN 50% PREFERENCE ON TEST 1 AND AN ADDITIONAL 9 WERE REMOVED FOR NOT MEETING HISTOLOGICAL CRITERIA)
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limitations[4]
for which uselimitation — the same scope as limitations[4], with the rule and its justification: WE DECIDED, A PRIORI, TO REMOVE ANIMALS THAT FAILED TO EXPRESS A PLACE PREFERENCE OF GREATER THAN 50% ON TEST 1
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limitations[5]
for which uselimitation — the strongest supporting evidence for the extinction claim, which is not shown: DESPITE DOUBLING THE NUMBER OF EXTINCTION TRIALS, WE FOUND THAT BOTH mPFC-INJECTED GROUPS CONTINUED TO EXHIBIT IMPAIRED EXTINCTION
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limitations[6]
for which uselimitation — the same scope as limitations[1] - the damage-to-projection account is offered as conceivable and explicitly not established
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limitations[7]
for which uselimitation — the central group contrast: THE INITIAL 2-WAY ANOVA REVEALED A MAIN EFFECT OF TEST [F(1,40) = 9.3, p < .005] BUT NOT GROUP, AND NO SIGNIFICANT GROUP x TEST INTERACTION (p's > .05), so the effect rests on the planned comparisons
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limitations[8]
replicationuntested
limitationThe planned within-subject comparisons were run regardless of the interaction and are acknowledged as not statistically justified.
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limitations[0]
limitationThe proposed explanation via damage to mPFC projection neurons is untested and needs confirmation.
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limitations[1]
limitationAs performed, the microinjection procedure cannot be used to test intra-mPFC pharmacological manipulations of EtOH-CPP extinction.
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limitations[2]
limitationThe authors state that their own analysis is not statistically justified, and say why they used it anyway
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limitations[3]
limitationAnimals were removed for failing to acquire a preference and for histology, about a third of each experiment
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limitations[4]
limitationNon-learners were excluded by an a priori rule, which is disclosed and defended by reference to prior practice
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limitations[5]
limitationThe strongest extra evidence - doubling extinction to six days - is reported as data not shown
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limitations[6]
limitationThe damage-to-projection account is offered as conceivable and explicitly not established
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limitations[7]
limitationNo group-by-test interaction reached significance in either experiment, so the key contrast rests on the planned within-group comparisons
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limitations[8]
Content tags
content-observations: finding · quantity · variable · definition · key-term — grounded in the full text
findingNeither the vehicle nor the bupivacaine group showed significant extinction of ethanol place preference.
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content_tags.findings[0]
findingIn the second experiment only the no-surgery group showed a significant drop in place preference.
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content_tags.findings[1]
findingThe experiments suggest some component of the microinjection procedure, not merely the handling, interfered with extinction.
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content_tags.findings[2]
findingDoubling the extinction trials did not rescue extinction: both injected groups kept showing place preference.
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content_tags.findings[3]
findingThe authors conclude the repeated intra-mPFC microinjection technique, as performed, is unsuitable for studying extinction of ethanol place preference in mice.
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content_tags.findings[4]
findingSurgery and anaesthesia alone are ruled out as the cause, since a single electrode insertion at the same coordinates in earlier work left extinction intact.
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content_tags.findings[5]
findingvehicle and bupivacaine groups both failed to extinguish
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content_tags.findings[6]
findingin the second experiment only the unoperated group extinguished
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content_tags.findings[7]
findinghandling alone does not explain it, since the no-surgery group was handled and still extinguished
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content_tags.findings[8]
findingthe tissue damage was substantial and limited mainly to layers I-III of the mPFC
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content_tags.findings[9]
findingdoubling the extinction training did not rescue it
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content_tags.findings[10]
findingsurgery and anaesthesia were ruled out by a matched earlier study in which a single electrode insertion did not impair extinction
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content_tags.findings[11]
quantityn=108 — total number of adult male DBA/2J mice used across the experiments
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content_tags.quantities[0]
quantityF (1,33) = 4.3, p < .05 — main effect of Test on percent time on the ethanol-paired floor in Experiment 1
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content_tags.quantities[1]
quantityF (1,40) = 9.3, p < .005 — main effect of Test on place preference in Experiment 2
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content_tags.quantities[2]
quantityt (11) = 2.5 p < .05 — the only significant within-group preference reduction, found in the No-Surgery group
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content_tags.quantities[3]
quantity18 — animals excluded from Experiment 1, split between failure to show preference and failed histology
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content_tags.quantities[4]
quantityless than 25% — share of subjects that failed to acquire a place preference after two conditioning trials
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content_tags.quantities[5]
quantity.05 — the alpha level adopted for all statistical tests
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content_tags.quantities[6]
quantity0.2 μL over 2 min (0.1 μL/min) — volume and rate of each intra-mPFC infusion of vehicle or bupivacaine
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content_tags.quantities[7]
quantityn=108 — total mice across both experiments before exclusions
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content_tags.quantities[8]
quantity18 animals removed — exclusions in Experiment 1 - nine non-learners and nine on histology
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content_tags.quantities[9]
quantity16 animals removed — the equivalent in Experiment 2
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content_tags.quantities[10]
quantity0.2 μL over 2 min — the injection volume and rate, chosen to inactivate most of the target subregions
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content_tags.quantities[11]
quantity>90% of neuronal activation — the intended coverage of the inactivation, from prior diffusion studies
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content_tags.quantities[12]
quantity2 g/kg — the ethanol dose paired with the floor cue
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content_tags.quantities[13]
variableNo Sx group (control)
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content_tags.variables[3]
variableinjector-tip location (control)
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content_tags.variables[9]
definitionbupivacaine
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content_tags.definitions[1]
definitionbupivacaine
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content_tags.definitions[2]
definitionextinction
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content_tags.definitions[3]
key-termsmedial prefrontal cortex, conditioned place preference, extinction, bupivacaine, microinjection, vehicle, prelimbic, infralimbic, guide cannula, lesion, nucleus accumbens, conditioned stimuli, histological criteria, conditioned fear
Statistics — cross-field join keys
field-invariant statistical terms, used in their statistical sense (gated on a number / symbol / test). A p-value or a standard deviation means the SAME thing in physics, biology, psychology and sociology — so these are the strongest cross-field join keys. Grounded verbatim.
ANOVA F as written: analysis of variance (ANOVA)
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content_tags.statistics[0]
two-way ANOVA with interaction term F as written: 2-way ANOVA (Group × Test)
Quotation withheld — this paper is not ours to reproduce, and the page has already quoted as much of it as the rights layer allows. The tag, its field and its confidence stand; the sentence is at the publisher.
content_tags.statistics[1]
paired t-test t as written: paired t-tests
Quotation withheld — this paper is not ours to reproduce, and the page has already quoted as much of it as the rights layer allows. The tag, its field and its confidence stand; the sentence is at the publisher.
content_tags.statistics[2]
Bonferroni correction for multiple comparisons as written: Bonferroni-adjusted within-subjects comparisons
Quotation withheld — this paper is not ours to reproduce, and the page has already quoted as much of it as the rights layer allows. The tag, its field and its confidence stand; the sentence is at the publisher.
content_tags.statistics[3]
alpha level / significance threshold α as written: Overall α was set at .05
Quotation withheld — this paper is not ours to reproduce, and the page has already quoted as much of it as the rights layer allows. The tag, its field and its confidence stand; the sentence is at the publisher.
content_tags.statistics[4]
standard error of the mean SEM as written: mean (± SEM)
Quotation withheld — this paper is not ours to reproduce, and the page has already quoted as much of it as the rights layer allows. The tag, its field and its confidence stand; the sentence is at the publisher.
content_tags.statistics[5]

When we have changed our mind. Some entries in the table begin with a word in capitals and the reason for it. EVIDENCE CORRECTED — the finding still stands, but we had quoted the wrong sentence from the paper, and the quotation has been replaced with the right one. RETRACTED — the entry itself was wrong; it stays on the page, marked, with the reason, rather than being deleted. WITHDRAWN AND REPLACED — the entry was wrong, and a corrected version of it appears in the entries below. Each mark carries its own reason, in brackets, because the reasons differ from one entry to the next. An entry we have WITHDRAWN is removed from this page and kept in the record, with its reason. A line reading Audit and a date is a later re-reading of the entry against its whole sentence: DOUBTFUL means it was kept, and the reason says what is loose about it. Quotation marks inside the tagger's sentence. Where a tag is the tagger's own sentence, a phrase in quotation marks inside it is borrowed from the paper — the tagger's wording around it is not. The paper's own sentence is the evidence quote for that row; on this page it is withheld with the article, and the article is at its publisher. Nothing inside quotation marks anywhere on this page is ours: if it is quoted, it is the paper's. Whose words a concept carries. A concept entry records a term the paper itself uses, so the term, the sense given for it, and its evidence are the paper's own words, quoted where the paper fixes them. This is the opposite of a sense entry in the tag table, which is the tagger's sentence about how the paper uses a term. Where a concept entry could not be grounded in the paper's words, the entry says so on its own row.

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