Prenatal heroin exposure alters cholinergic receptor stimulated activation of the PKCbetaII and PKCgamma isoforms
Authors: Zvi Naor
Joseph Yanai
Shiri P Yaniv
Journal: Brain Research Bulletin (2004)
Discipline: NEUROSCIENCE & BEHAVIOR
Keywords: Prenatal exposure; heroin; behavioral deficits; cholinergic innervation; PKC activity
Affiliations: The Ross Laboratory for Studies in Neural Birth Defects, Department of Anatomy and Cell Biology, The Hebrew University-Hadassah Medical School, Box 12272, Jerusalem 91120, Israel.

Abstract:  Prenatal exposure of mice to heroin (SC injection of 10mg/kg to the dams on gestational days 9-18) resulted at adulthood in behavioral deficits related to septohippocampal cholinergic innervation accompanied with both presynaptic and postsynaptic cholinergic hyperactivity; including an increase membrane PKC activity, and a desensitization of PKC to cholinergic input which were highly correlated with the behavioral performance and were reversed by cholinergic grafting. Therefore, we studied the receptor induced activation of the behaviorally relevant PKCgamma and PKCbetaII isoforms and the less behaviorally relevant PKCalpha isoform. Time course studies revealed peak translocation after 40 min incubation with carbachol for PKCgamma (110% increase from basal, i.e. no carbachol level, P &lt; 0.01), 30 min for phosphorylated PKCbetaII (130%, P &lt; 0.05) and 5 min for non-phosphorylated PKCbetaII (64%, P &lt; 0.05) with no peak for alpha. Prenatal heroin abolished the translocation of PKCgamma and PKCbetaII while PKCalpha remained unaffected. A decrease occurred in basal phosphorylated membrane (-45%, P &lt; 0.01) and cytosol-associated (-29%, P &lt; 0.01) PKCbetaII, in membrane-associated non-phosphorylated PKCbetaII (-32%, P &lt; 0.01) and PKCgamma (-25%, P &lt; 0.01) and in cytosolic PKCalpha (-27%, P &lt; 0.01), while membrane-associated PKCalpha was slightly increased (11%, P &lt; 0.05). The results suggest that prenatal heroin disrupts cholinergic receptor induced PKC translocation and activation with the underlying mechanism of neuroteratogenicity potentially lying in the PKCgamma and PKCbetaII, while PKCalpha remains unaffected. 

Retracted: No

PubPeer comments:
 2004 was the year when lane splicing was officially deprecated. https: pubmed.ncbi.nlm.nih.gov 15240566 It had become clear that splicing sundry lanes together meant that it was impossible to tell whether the experiment had been done as described - or if the splicing was a symptom of deeper issues. But when it comes to cutting and pasting bands into gellish backgrounds, well was that ever an acceptable practice? At first, I feared the opioids were obfuscating my visual deconstruction capabilities. But, seen in the cold light of day, in this 2004 Brain Research Bulletin there are a surprising number of bands pasted into what are, Condy understands, figure images purporting to be western blots. Let us begin with Fig. 1F, original greyscale and as transmogrified by FIJI LUT ![image] , 
 I had nothing to do with the Ms preparation. My only role was to provide ideas and some reagents. , 
 #2 An entirely plausible contribution for a PI as middle author. But was an opportunity missed to pass an eye over the figures in the draft manuscript before submission? Fig. 3C has band pasting superposed on background pasting in lanes 1,2. How very mysterious. ![image] , 
 I saw the draft and the figures and thought they were fine and ready for publication , 
 #4 On the face of it, to think these figures were ready for publication does not make any sense. These blottish images are, to put it in a clear and simple way, absolutely awful. As in: truly, madly, deeply awful. How could a coauthor inspect them and not notice that anything was awry? But yet - might I seem to remember that Wordly manuscripts being bandied around in 2004 could have astonishingly low resolution for their embedded figures? Perhaps the worthy goal of the given software supplier was to minimise file size and costly bandwidth on the somewhat rudimentary intarwebs? So I could imagine that supplied artifactually granular imagery might not have been as transparent to a coauthor reading the contemporary draft as it is to those of us who choose to eyeball the published figures with full on 2022 vision deployed in hindsight. Figure 2D shows both vertical and horizontal assembly edges, suggesting that this particular WB never existed ![image] , 
 As I said before; I saw the draft figures and thought they were fine (Z.N) , 
 #6 I like to think that, had I been a coauthor, I would have noticed the artefacts and not signed off on these images. But of course hindsight is a wonderful thing so I should not get too presumptuous. In the Figure 5B WB, it looks as though the rubber tool has been applied between the two lanes, presumably to smooth out the splice edge. Note the different grey scales of the two lanes and the loss of background granularity between them. ![image] , 
 When you enlarge a figure to such a degree you see what you want to see sorry ,