Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting
IntroductionIn both rats and humans, methamphetamine (Meth) seeking progressively increases during abstinence, a behavioral phenomenon termed “incubation of Meth craving”. We previously demonstrated a critical role of dorsal striatum (DS) in this incubation in rats. However, circuit-specific molecul...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-02-01
|
| Series: | Frontiers in Cellular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fncel.2025.1542508/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1823856600340758528 |
|---|---|
| author | Rachel D. Altshuler Megan A. M. Burke Kristine T. Garcia Kenneth Class Raffaello Cimbro Xuan Li Xuan Li |
| author_facet | Rachel D. Altshuler Megan A. M. Burke Kristine T. Garcia Kenneth Class Raffaello Cimbro Xuan Li Xuan Li |
| author_sort | Rachel D. Altshuler |
| collection | DOAJ |
| description | IntroductionIn both rats and humans, methamphetamine (Meth) seeking progressively increases during abstinence, a behavioral phenomenon termed “incubation of Meth craving”. We previously demonstrated a critical role of dorsal striatum (DS) in this incubation in rats. However, circuit-specific molecular mechanisms in DS underlying this incubation are largely unknown. Here we combined a newly developed fluorescence-activated sorting (FACS) protocol with fluorescence-conjugated cholera toxin subunit B-647 (CTb-647, a retrograde tracer) to examine gene alterations in the direct-pathway (striatonigral) medium spiny neurons (MSNs) associated with incubation of Meth craving.MethodsWe injected CTb-647 bilaterally into substantia nigra before or after training rats to self-administer Meth or saline (control condition) for 10 days (6 h/d). On abstinence day 1 or day 28, we collected the DS tissue from both groups for subsequent FACS and examined gene expressions in CTb-positive (striatonigral MSNs) and CTb-negative (primarily non-striatonigral MSNs). Finally, we examined gene expressions in DS homogenates, to demonstrate cell-type specificity of gene alterations observed on abstinence day 28.ResultsOn abstinence day 1, we found mRNA expression of Gabrb3 decreased only in CTb-positive (but not CTb-negative) neurons of Meth rats compared with saline rats, while mRNA expression of Usp7 decreased in all sorted DS neurons. On abstinence day 28, we found increased mRNA expression for Grm3, Opcml, and Usp9x in all sorted DS neurons, but not DS homogenate.DiscussionTogether, these data demonstrated that incubation of Meth craving was associated with time-dependent, circuit-specific, and cell type-specific gene alterations in DS involved in glutamatergic, GABAergic, opioidergic, and protein degradation signaling. |
| format | Article |
| id | doaj-art-8801e4088db5496cbffe084f19aad413 |
| institution | Kabale University |
| issn | 1662-5102 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj-art-8801e4088db5496cbffe084f19aad4132025-02-12T07:25:54ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022025-02-011910.3389/fncel.2025.15425081542508Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sortingRachel D. Altshuler0Megan A. M. Burke1Kristine T. Garcia2Kenneth Class3Raffaello Cimbro4Xuan Li5Xuan Li6Department of Psychology, University of Maryland College Park, College Park, MD, United StatesDepartment of Psychology, University of Maryland College Park, College Park, MD, United StatesDepartment of Psychology, University of Maryland College Park, College Park, MD, United StatesDepartment of Cell Biology and Molecular Genetics, University of Maryland College Park, College Park, MD, United StatesDivision of Rheumatology, Johns Hopkins University School of Medicine, Baltimore, MD, United StatesDepartment of Psychology, University of Maryland College Park, College Park, MD, United StatesProgram in Neuroscience and Cognitive Science, University of Maryland College Park, College Park, MD, United StatesIntroductionIn both rats and humans, methamphetamine (Meth) seeking progressively increases during abstinence, a behavioral phenomenon termed “incubation of Meth craving”. We previously demonstrated a critical role of dorsal striatum (DS) in this incubation in rats. However, circuit-specific molecular mechanisms in DS underlying this incubation are largely unknown. Here we combined a newly developed fluorescence-activated sorting (FACS) protocol with fluorescence-conjugated cholera toxin subunit B-647 (CTb-647, a retrograde tracer) to examine gene alterations in the direct-pathway (striatonigral) medium spiny neurons (MSNs) associated with incubation of Meth craving.MethodsWe injected CTb-647 bilaterally into substantia nigra before or after training rats to self-administer Meth or saline (control condition) for 10 days (6 h/d). On abstinence day 1 or day 28, we collected the DS tissue from both groups for subsequent FACS and examined gene expressions in CTb-positive (striatonigral MSNs) and CTb-negative (primarily non-striatonigral MSNs). Finally, we examined gene expressions in DS homogenates, to demonstrate cell-type specificity of gene alterations observed on abstinence day 28.ResultsOn abstinence day 1, we found mRNA expression of Gabrb3 decreased only in CTb-positive (but not CTb-negative) neurons of Meth rats compared with saline rats, while mRNA expression of Usp7 decreased in all sorted DS neurons. On abstinence day 28, we found increased mRNA expression for Grm3, Opcml, and Usp9x in all sorted DS neurons, but not DS homogenate.DiscussionTogether, these data demonstrated that incubation of Meth craving was associated with time-dependent, circuit-specific, and cell type-specific gene alterations in DS involved in glutamatergic, GABAergic, opioidergic, and protein degradation signaling.https://www.frontiersin.org/articles/10.3389/fncel.2025.1542508/fullmethamphetamine cravingcholera toxin subunit Bfluorescence-activated cell sortingstriatonigral projection neuronsgene expression |
| spellingShingle | Rachel D. Altshuler Megan A. M. Burke Kristine T. Garcia Kenneth Class Raffaello Cimbro Xuan Li Xuan Li Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting Frontiers in Cellular Neuroscience methamphetamine craving cholera toxin subunit B fluorescence-activated cell sorting striatonigral projection neurons gene expression |
| title | Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting |
| title_full | Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting |
| title_fullStr | Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting |
| title_full_unstemmed | Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting |
| title_short | Profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit B-based fluorescence-activated cell sorting |
| title_sort | profiling gene alterations in striatonigral neurons associated with incubation of methamphetamine craving by cholera toxin subunit b based fluorescence activated cell sorting |
| topic | methamphetamine craving cholera toxin subunit B fluorescence-activated cell sorting striatonigral projection neurons gene expression |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2025.1542508/full |
| work_keys_str_mv | AT racheldaltshuler profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting AT meganamburke profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting AT kristinetgarcia profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting AT kennethclass profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting AT raffaellocimbro profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting AT xuanli profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting AT xuanli profilinggenealterationsinstriatonigralneuronsassociatedwithincubationofmethamphetaminecravingbycholeratoxinsubunitbbasedfluorescenceactivatedcellsorting |