The Cerebellum on Cocaine

The traditional cerebellum’s role has been linked to the high computational demands for sensorimotor control. However, several findings have pointed to its involvement in executive and emotional functions in the last decades. First in 2009 and then, in 2016, we raised why we should consider the cerebellum when thinking about drug addiction. A decade later, mounting evidence strongly suggests the cerebellar involvement in this disorder. Nevertheless, direct evidence is still partial and related mainly to drug-induced reward memory, but recent results about cerebellar functions may provide new insights into its role in addiction. The present review does not intend to be a compelling revision on available findings, as we did in the two previous reviews. This minireview focuses on specific findings of the cerebellum’s role in drug-related reward memories and the way ahead for future research. The results discussed here provide grounds for involving the cerebellar cortex’s apical region in regulating behavior driven by drug-cue associations. They also suggest that the cerebellar cortex dysfunction may facilitate drug-induced learning by increasing glutamatergic output from the deep cerebellar nucleus (DCN) to the ventral tegmental area (VTA) and neural activity in its projecting areas

The role of the cerebellum in drug-cue associative memory: functional interactions with the medial prefrontal cortex

Drug-induced Pavlovian memories are thought to be crucial for drug addiction because they guide behaviour towards environments with drug availability. Drug-related memory depends on persistent changes in dopamine-glutamate interactions in the medial pre- frontal cortex (mPFC), basolateral amygdala, nucleus accumbens core and hippocampus. Recent evidence from our laboratory indi- cated that the cerebellum is also a relevant node for drug-cue associations. In the present study, we tested the role that specific regions of the cerebellum and mPFC play in the acquisition of cocaine-induced preference conditioning. Quinolinic acid was used to manage a permanent deactivation of lobule VIII in the vermis prior to conditioning. Additionally, lidocaine was infused into the prelim- bic and infralimbic (IL) cortices for reversible deactivation before every training session. The present findings show, for the first time, that the cerebellum and mPFC might act together in order to acquire drug-cue Pavlovian associations. Either a dorsal lesion in lobule VIII or an IL deactivation encouraged cocaine-induced preference conditioning. Moreover, simultaneous IL-cerebellar deactivation prevented the effect of either of the separate deactivations. Therefore, similar to the IL cortex, neural activity in the cerebellum may be crucial for ensuring inhibitory control of the expression of cocaine-related memories

Mapping cFos expression after cerebellar and medial prefrontal deactivations in rats trained to acquire cocaine-induced preference conditioning

6th Mediterranean Conference of Neuroscience (MNS Malta 2017

Cocaine-Induced Preference Conditioning: a Machine Vision Perspective

Existing work on drug-induced synaptic changes has shown that the expression of perineuronal nets (PNNs) at the cerebellar cortex can be regulated by cocaine-related memory. However, these studies on animals have mostly relied on limited manually-driven procedures, and lack some more rigorous statistical approaches and more automated techniques. In this work, established methods from computer vision and machine learning are considered to build stronger evidence of those previous findings. To that end, an image descriptor is designed to characterize PNNs images; unsupervised learning (clustering) is used to automatically find distinctive patterns of PNNs; and supervised learning (classification) is adopted for predicting the experiment group of the mice from their PNN images. Experts in neurobiology, who were not aware of the underlying computational procedures, were asked to describe the patterns emerging from the automatically found clusters, and their descriptions were found to align surprisingly well with the two types of PNN images revealed from previous studies, namely strong and weak PNNs. Furthermore, when the set of PNN images corresponding to every mice in the saline (control) group and the conditioned (experimental) group were characterized using a bag-of-words representation, and subject to supervised learning (saline vs conditioned mice), the high classification results suggest the ability of the proposed representation and procedures in recognizing these groups. Therefore, despite the limited size of the dataset (1,032 PNN images of 6 saline and 6 conditioned mice), the results support existing evidence on the drug-related brain plasticity, while providing higher objectivit

A Working Hypothesis for the Role of the Cerebellum in Impulsivity and Compulsivity

Growing evidence associates cerebellar abnormalities with several neuropsychiatric disorders in which compulsive symptomatology and impulsivity are part of the disease pattern. Symptomatology of autism, addiction, obsessive-compulsive (OCD), and attention deficit/hyperactivity (ADHD) disorders transcends the sphere of motor dysfunction and essentially entails integrative processes under control of prefrontal-thalamic-cerebellar loops. Patients with brain lesions affecting the cortico-striatum thalamic circuitry and the cerebellum indeed exhibit compulsive symptoms. Specifically, lesions of the posterior cerebellar vermis cause affective dysregulation and deficits in executive function. These deficits may be due to impairment of one of the main functions of the cerebellum, implementation of forward internal models of the environment. Actions that are independent of internal models may not be guided by predictive relationships or a mental representation of the goal. In this review article, we explain how this deficit might affect executive functions. Additionally, regionalized cerebellar lesions have been demonstrated to impair other brain functions such as the emergence of habits and behavioral inhibition, which are also altered in compulsive disorders. Similar to the infralimbic cortex, clinical studies and research in animal models suggest that the cerebellum is not required for learning goal-directed behaviors, but it is critical for habit formation. Despite this accumulating data, the role of the cerebellum in compulsive symptomatology and impulsivity is still a matter of discussion. Overall, findings point to a modulatory function of the cerebellum in terminating or initiating actions through regulation of the prefrontal cortices. Specifically, the cerebellum may be crucial for restraining ongoing actions when environmental conditions change by adjusting prefrontal activity in response to the new external and internal stimuli, thereby promoting flexible behavioral control. We elaborate on this explanatory framework and propose a working hypothesis for the involvement of the cerebellum in compulsive and impulsive endophenotypes

Plasticity in the infralimbic cortex after a lesion of the dorsal posterior cerebellum: an inhibitory pathway for drug-dependent memories

Treball final de Màster Universitari en Investigació en Cervell i Conducta. Codi: SBM024. Curs acadèmic 2015-2016Addiction could be considered the result of pathological learning. Drug-associated cues become strong conditioned stimuli that create drug-related memories which promote drug seeking leading to relapse. Both, the striatumcortico-limbic circuitry and the cerebellum store information about reinforcing stimuli and their related cues, sharing widespread connections. Previous studies from the lab showed that both, the IL and the cerebellum exhibit hallmark signatures of drug-induced memories, changes in c- Fos expression and in PNNs. Moreover, the deactivation of the infralimbic cortex (IL) promote preference towards cocaine-associated cues, and expressed increased c-Fos+ neurons, and stronger PNNs in Golgi neurons in the apex of lobule VIII of the cerebellum. In the present study, we managed the effect of a permanent lesion of the lobule VIII vermis of the cerebellum on cocaine-induced odour preference conditioning and plasticity changes in the IL. We aimed to elucidate whether the plasticity changes of drug-induced memories observed in the cerebellum can also be found in the IL. Our results showed that a permanent lesion of the lobule VIII in the vermis promotes preference towards cocaine-associated cues, an increased number of c-Fos+ neurons, and a reduction in the number of infralimbic neurons that expressed a PNN, but no differences were found between groups related to average intensity of the matrix expression. It seems that the IL cortex and vermis are part of a functional and structural network for inhibitory effects on the acquisition of these drug-dependent preference memories

Functional interactions of Infralimbic-Cerebellum pathways in cocaine-induced preference memory

Doctorat internacionalThe present thesis investigates the cerebellum-IL interactions in drug-induced memories and their anatomical pathways. To do so, we 1) assessed activity and perineuronal nets (PNNs) expression in the posterior cerebellum of IL-inactivated rats, 2) digested PNNs in LVIII at different time points of the learning process, and 3) created an anatomical map of the ascending and descending cerebellum-IL reciprocal pathways using tracing agents. First, IL inactivation promoted cocaine-induced memory acquisition and upregulated cFos and vGluT2 activity around neurogranin+ Golgi interneurons and PNN expression in the posterior cerebellum. Second, LVIII PNNs enzymatic digestion disrupted short-term cocaine memory and facilitated reinstatement of cocaine-induced memory probably by preventing the consolidation of extinction. Third, our anatomical maps show that LVIII via the interposed DCN might contact VTA and then contact IL, and IL might send projections to both interposed DCN and inferior olive and these areas that reach LVIII.Programa de Doctorat en Ciències Biomèdiques i Salu

Putting forward a model for the role of the cerebellum in cocaine-induced pavlovian memory

Substance Use Disorder (SUD) involves emotional, cognitive, and motivational dysfunction. Long-lasting molecular and structural changes in brain regions functionally and anatomically linked to the cerebellum, such as the prefrontal cortex, amygdala, hippocampus, basal ganglia, and ventral tegmental area, are characteristic of SUD. Direct and indirect reciprocal connectivity between the cerebellum and these brain regions can explain cerebellar roles in Pavlovian and reinforcement learning, fear memory, and executive functions. It is increasingly clear that the cerebellum modulates brain functions altered in SUD and other neuropsychiatric disorders that exhibit comorbidity with SUD. In the present manuscript, we review and discuss this evidence and present new research exploring the role of the cerebellum in cocaine-induced conditioned memory using chemogenetic tools (designer receptor exclusively activated by designer drug, DREADDs). Our preliminary data showed that inactivation of a region that includes the interposed and lateral deep cerebellar nuclei reduces the facilitating effect of a posterior vermis lesion on cocaine-induced preference conditioning. These findings support our previous research and suggest that posterior vermis damage may increase drug impact on the addiction circuitry by regulating activity in the DCN. However, they raise further questions that will also be discussed

Time-dependent regulation of perineuronal nets in the cerebellar cortex during abstinence of cocaine-self administration

International audienceRationale: The probability of structural remodeling in brain circuits may be modulated by molecules of perineuronal nets (PNNs) that restrict neuronal plasticity to stabilize circuits. Animal research demonstrates that addictive drugs can remodel PNNs in different brain regions, including the cerebellum.Objective: This study aimed to investigate the effects of short versus extended access to cocaine self-administration on PNN expression around Golgi interneurons in the cerebellar cortex after different periods of abstinence.Methods: After 1 week of training (2 h/day), Sprague-Dawley rats self-administered cocaine daily for 20 days under short (ShA) or extended (LgA) access. PNN expression in the cerebellum was assessed after 1 day, 7 days, and 28 days of forced abstinence. PNNs were immunolabeled using Wisteria floribunda agglutinin (WFA) and captured by confocal microscopy.Results: WFA intensity increased in PNN-bearing Golgi neurons over the abstinence period and a higher proportion of more intense PNNs were formed throughout the first month of abstinence. After the first 24 h of cocaine abstinence, however, we found a reduction in WFA intensity in the cerebellar cortex of rats with ShA to cocaine as compared to naïve animals. When comparing with naïve rats, LgA rats showed consistent PNN upregulation at 28 days of cocaine abstinence.Conclusions: Our results suggest that cocaine self-administration produces modifications in PNN that enhance conditions for synaptic plasticity in the cerebellar cortex. These modifications are revealed shortly after the cessation of drug intake but PNNs become more intense during protracted abstinence in the LgA group, pointing to the stabilization of drug-induced synaptic changes. These findings indicate that extended access to cocaine self-administration dynamically regulates conditions for plasticity in the cerebellum during abstinence

Role of Perineuronal nets in the cerebellar cortex in cocaine-induced conditioned preference, extinction, and reinstatement

Perineuronal nets (PNNs) are cartilage-like structures of extracellular matrix molecules that enwrap in a net-like manner the cell-body and proximal dendrites of special subsets of neurons. PNNs stabilize their incoming connections and restrict plasticity. Consequently, they have been proposed as a candidate mechanism for drug-induced learning and memory. In the cerebellum, PNNs surround Golgi inhibitory interneurons and both inhibitory and excitatory neurons in the deep cerebellar nuclei (DCN). Previous studies from the lab showed that cocaine-induced conditioned memory increased PNN expression in the granule cell layer of the posterior vermis. The present research aimed to investigate the role of cerebellar PNNs in cocaine-induced conditioned preference. For this purpose, we use the enzyme chondroitinase ABC (ChABC) to digest PNNs at different time points of the learning process to ascertain whether their removal can affect drug-induced memory. Our results show that PNN digestion using ChABC in the posterior vermis (Lobule VIII) did not affect the acquisition of cocaine-induced conditioned preference. However, the removal of PNNs in Lobule VIII -but not in the DCN- disrupted short-term memory of conditioned preference. Moreover, although PNN digestion facilitated the formation of extinction, reinstatement of cocaine-induced conditioned preference was encouraged under PNN digestion. The present findings suggests that PNNs around Golgi interneurons are needed to maintain cocaine-induced Pavlovian memory but also to stabilize extinction memory. Conversely, PNN degradation within the DCN did not affect stability of cocaine-induced memories. Therefore, degradation of PNNs in the vermis might be used as a promising tool to manipulate drug-induced memory.Funding for open access charge: CRUE-Universitat Jaume