Escalated Oxycodone Self-Administration Causes Differential Striatal mRNA Expression of FGFs and IEGs Following Abstinence-Associated Incubation of Oxycodone Craving

Blackwood, Christopher A.; Leary, Michael; Salisbury, Aaron J.; McCoy, Michael T.; Cadet, Jean Lud

doi:10.1016/j.neuroscience.2019.07.030

Cited by 33 publications

(36 citation statements)

References 66 publications

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“…This idea is supported by our recent demonstration that decreases in the expression of Mu opioid receptor protein levels in the dorsal striatum correlated with escalated drug intake and drug seeking behavior in oxycodone self-administering rats (Blackwood et al, 2018). This reasoning is also consonant with our findings of dose-dependent changes in the mRNA expression of immediate early genes (IEGs) following oxycodone seeking after a month of withdrawal from escalated oxycodone SA (Blackwood et al, 2019). We reasoned further that shock-resistant and shock-sensitive rats may show differential expression of opioid receptors in the dorsal striatum and of IEGs in both the dorsal striatum and PFC.…”

Section: Introductionsupporting

confidence: 80%

“…Therefore, we had predicted that the SR rats might show acute increased expression of some IEGs because they were taking oxycodone just 2-h before they were euthanized. We had, indeed, previously found increased expression of c-fos in the dorsal striatum of rats that were euthanized after 1 month of withdrawal from escalated oxycodone intake (Blackwood et al, 2019). We also thought that the levels of expression of IEGs might be normal or might be reduced in the SS group in comparison to the SR rats.…”

Section: Discussionmentioning

confidence: 83%

“…Total RNA (0.5 µg) was reverse-transcribed (RT) with oligo dT primers using Advantage RT-for-PCR kit (Clontech, Mountain View, CA, United States). RT-qPCR was performed as previously described (Cadet et al, 2017;Blackwood et al, 2019) with Roche LightCycler 480 II (Roche Diagnostics, Indianapolis, IN, United States) using iQ SYBR Green Supermix (Bio-Rad, Hercules, CA, United States). Custom-designed primers were made and HPLC-purified by the Synthesis and Sequence Facility of the Johns Hopkins University (Baltimore, MD, United States).…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

“…As noted in the introduction, because the expression of several IEGs is known to be impacted by administration of opioid drugs (Bisagno and Cadet, 2019;Blackwood et al, 2019), we reasoned that differential expression of some IEGs of the Fos family might occur in the SR and SS oxycodone SA phenotypes, with the SR showing higher mRNA expression. Figure 3 shows the effects of footshocks and oxycodone SA on c-fos, fosB, N), and fra2 (O,P) mRNA levels were detected in the PFC.…”

Section: Dorsal Striatummentioning

confidence: 99%

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Escalated Oxycodone Self-Administration and Punishment: Differential Expression of Opioid Receptors and Immediate Early Genes in the Rat Dorsal Striatum and Prefrontal Cortex

Blackwood¹,

McCoy²,

Ladenheim³

et al. 2020

Front. Neurosci.

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Opioid use disorder (OUD) is characterized by compulsive drug taking despite adverse life consequences. Here, we sought to identify neurobiological consequences associated with the behavioral effects of contingent footshocks administered after escalation of oxycodone self-administration. To reach these goals, Sprague-Dawley rats were trained to self-administer oxycodone for 4 weeks and were then exposed to contingent electric footshocks. This paradigm helped to dichotomize rats into two distinct behavioral phenotypes: (1) those that reduce lever pressing (shock-sensitive) and (2) others that continue lever pressing (shock-resistant) for oxycodone during contingent punishment. The rats were euthanized at 2-h after the last oxycodone plus footshock session. The dorsal striata and prefrontal cortices were dissected for use in western blot and RT-qPCR analyses. All oxycodone self-administration rats showed significant decreased expression of Mu and Kappa opioid receptor proteins only in the dorsal striatum. However, expression of Delta opioid receptor protein was decreased in both brain regions. RT-qPCR analyses documented significant decreases in the expression of c-fos, fosB, fra2, junB, egr1, and egr2 mRNAs in the dorsal striatum (but not in PFC) of the shock-sensitive rats. In the PFC, junD expression was reduced in both phenotypes. However, egr3 mRNA expression was increased in the PFC of only shock-resistant rats. These results reveal that, similar to psychostimulants and alcohol, footshocks can dichotomize rats that escalated their intake of oxycodone into two distinct behavioral phenotypes. These animals also show significant differences in the mRNA expression of immediate early genes, mainly, in the dorsal striatum. The increases in PFC egr3 expression in the shock-resistant rats suggest that Egr3 might be involved in the persistence of oxycodone-associated memory under aversive conditions. This punishment-driven model may help to identify neurobiological substrates of persistent oxycodone taking and abstinence in the presence of adverse consequences.

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Section: Introductionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 83%

Section: Quantitative Rt-pcrmentioning

confidence: 99%

Section: Dorsal Striatummentioning

confidence: 99%

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Escalated Oxycodone Self-Administration and Punishment: Differential Expression of Opioid Receptors and Immediate Early Genes in the Rat Dorsal Striatum and Prefrontal Cortex

Blackwood¹,

McCoy²,

Ladenheim³

et al. 2020

Front. Neurosci.

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“…Size measurements and neurosphere counts were analyzed in Adobe Photoshop (version 10.0.1) CS3 Extended (Adobe; San Jose, CA) using the measurement function (ImageJ is an open-access alternative that can be used to perform this function). Statistical analyses were carried out as previously described 12,13…”

Section: Data Acquisition and Statisticsmentioning

confidence: 99%

Quantitative approach to numbers and sizes: Generation of primary neurospheres from the dorsal lateral ganglionic eminence of late embryonic mice

Blackwood

2020

F1000Res

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Background: The neurosphere assay is a powerful in vitro tool to investigate neural stem cells in the dorsal lateral ventricle (dLGE). In the dLGE, metrics of sizes and numbers of neurospheres generated using this assay has not been completely characterized. The objective of this protocol is to provide a stepwise method from a single isolation that predicts the average number of neurospheres generated and to estimate an approximation of its sizes after several days in vitro. The advantage of this protocol is that no expensive and specialized equipment is needed for tissue isolation. Estimates about the numbers and sizes of neurospheres will provide investigators with quantitative data to advise on how much starting dLGE tissue is required to generate the appropriate number of spheres for the implementation of downstream applications, including immunocytochemistry, self-renewal and differentiation assays. Methods: Our method is based on a simple dissection technique, where tissue surrounding the dorsal lateral ventricle from a single mouse embryo is trimmed away to enrich for neural stem cell and progenitor populations. Following this dissection, tissue is mechanically dissociated by trituration. Cells are then cultured in media containing epidermal growth factor and other supplements to generate healthy primary neurospheres. Results: Using this approach, we found reproducible number of primary neurospheres after 7 days in vitro (DIV). Furthermore, we observed that this method yields an average range of neurospheres sizes greater than 50 μm, but less than 100 μm after 7 DIV. Lastly, using an anti-GFAP antibody, we show that these neurospheres can be stained, confirming their use in future immunocytochemistry studies. Conclusions: Future use of this protocol provides metrics on the generation of primary neurospheres that will be useful for further advances in the area of stem cell biology.

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Electrochemical sensor based on glassy carbon electrode modified by polymelamine formaldehyde/graphene oxide nanocomposite for ultrasensitive detection of oxycodone

et al. 2021

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Escalated Oxycodone Self-Administration Causes Differential Striatal mRNA Expression of FGFs and IEGs Following Abstinence-Associated Incubation of Oxycodone Craving

Cited by 33 publications

References 66 publications

Escalated Oxycodone Self-Administration and Punishment: Differential Expression of Opioid Receptors and Immediate Early Genes in the Rat Dorsal Striatum and Prefrontal Cortex

Escalated Oxycodone Self-Administration and Punishment: Differential Expression of Opioid Receptors and Immediate Early Genes in the Rat Dorsal Striatum and Prefrontal Cortex

Quantitative approach to numbers and sizes: Generation of primary neurospheres from the dorsal lateral ganglionic eminence of late embryonic mice

Electrochemical sensor based on glassy carbon electrode modified by polymelamine formaldehyde/graphene oxide nanocomposite for ultrasensitive detection of oxycodone

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