Name: Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference (Video) | JoVE Journal
Uploaded: 2013-07-30T12:00:00.000+00:00
Duration: 8 min 22 s
Description: Learn stereotaxic microinjection of viral vectors expressing Cre recombinase to study gene function in cocaine conditioned place preference using mice. This method enables precise gene knockout during acquisition, extinction, and reinstatement phases.

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5 minutes

The authors would like to thank Anni Lee and Maureen Byrne for their help in establishing the extended conditioned place preference protocol.

All procedures are conducted in accordance with the Weill Cornell Medical College Institutional Animal Care and Use Committee rules.
1. Preparation and Setup for Stereotaxic Delivery of Viral Vectors
<ol>
	<li>If an instrument, sterile swab, or hand wearing sterile glove is contaminated by coming into contact with a non-sterile surface, discard or re-sterilize the instrument using a hot bead sterilizer, discard the swab or change to new sterile gloves.</li>
	<li>Place a mouse cage with bedding on a heat block to warm for post-operative recovery.</li>
	<li>Set up electric razor for shaving head and ethanol and iodine to sterilize the scalp.</li>
	<li>Wipe down ear bars and mouth hold of the stereotax with 70% ethanol.</li>
	<li>Using a 5 &mu;l Hamilton syringe, draw up 5 &mu;l of viral vector (1 x 106 viral particles/&mu;l) and set up in syringe holder on stereotax.</li>
	<li>Microwave heating pad or turn electric pad on and place on stereotax. Cover with a clean absorbent pad to reduce the risk of burns. Monitor mouse body temperature (between 97-99.5 &deg;F) frequently throughout procedure. </li>
	<li>Inject mouse (weighing between 22-35 g) with a mixture of ketamine (100 mg/kg) and xylazine (20 mg/kg). Once a mouse is anesthetized, pinch tail tip to check the mouse's reflexes and note any movement or response to ensure that the mouse is adequately anesthetized.</li>
	<li>Shave head between ears and eyes.</li>
	<li>Clean shaved head with alternating ethanol and iodine sterile cotton applicators.</li>
	<li>Transfer mouse to heated pad on stereotax.</li>
</ol>
2. Stereotaxic Microinjection of Viral Vectors
<ol>
	<li>Wear sterile gloves throughout the surgery. If gloves are contaminated at any point by touching non-sterile surfaces, change gloves.</li>
	<li>Zero tooth bar and ear bar scales.</li>
	<li>Set teeth in tooth bar.</li>
	<li>Screw in muzzle. If wiggling or whisker movement is noted at any time, administer an i.p. booster dose (~0.05 cc) of a mixture of ketamine (100 mg/kg) and xylazine (20 mg/kg).</li>
	<li>Screw in ear bars.</li>
	<li>Lubricate eyes with PuraLube.</li>
	<li>Cut open scalp to just behind ears with a scalpel.</li>
	<li>Place bulldog clips on the corners of the scalp to peel the skin away from skull. Bregma and lambda should be easily visible.</li>
	<li>Make sure suture from bregma to lambda is aligned straight. If the suture is crooked readjust positioning of the head.</li>
	<li>Check to make sure all settings on the stereotax are zeroed.</li>
	<li>Put needle tip in place at bregma.</li>
	<li>Measure all coordinates for both bregma and lambda. This includes dorsal/ventral, medial/lateral and anterior/posterior coordinates.</li>
	<li>Adjust the head until the ventral coordinates for bregma and lambda are within 0.02 mm of each other.</li>
	<li>Once aligned, from bregma go medially out to the medial/lateral (M/L) coordinates. Make sure the left and right sides are aligned within 0.02 mm of each other.</li>
	<li>Re-center needle tip on bregma and from there move in the anterior/posterior (A/P) direction to the desired anterior/posterior coordinate.</li>
	<li>If necessary, angle needle in one direction and make sure the stereotax is locked. Go out to the left M/L coordinate (+/- 1.52)</li>
	<li>Measure the ventral position of the top of the skull to use as a reference after the borehole is drilled.</li>
	<li>Drill a borehole using a sterile drill bit at the desired coordinate. Check that the needle can enter uninterrupted through the skull.</li>
	<li>Lower the needle to the desired ventral coordinate.</li>
	<li>Once the coordinate is reached, dip 0.015 mm below for ~10 sec to create a small pocket for the viral vector.</li>
	<li>Inject the desired volume of viral vector at a rate of ~0.1 &mu;l/min. Wait 3 min for viral vector to diffuse completely. Raise the needle tip 0.015 mm and wait an additional 2 min.</li>
	<li>Dip the needle (if doing angled injections) in the opposite direction. Again, record the ventral position as a reference for the top of the skull.</li>
	<li>Repeat the same steps as on the first side to inject viral vector on the opposite side (if doing bilateral injections).</li>
	<li>Apply bone wax to both holes to seal them using the wooden end of a sterile cotton applicator tip.</li>
	<li>Suture the scalp.</li>
	<li>Apply nerve block to the wounded area.</li>
	<li>Remove mouse from stereotax and place in warmed cage on heat block to recover for 45 min.</li>
	<li>Monitor for signs of pain including: decreased overall activity or restlessness, decreased food and water consumption or increased vocalization. Buprenorphine (0.05 - 0.2 mg/kg) can be administered twice daily for a week if necessary for pain. Acceptable alternatives include: meperidine (20-60 mg/kg), pentazocine (10 mg/kg), nalbuphine (4-8 mg/kg).</li>
</ol>
3. Conditioned Place Preference: Acquisition, Extinction, and Reinstatement
<ol>
	<li>Place mice into the central chamber of a three-chamber place preference apparatus (Med Associates Inc., St. Albans, VT, USA) for a 60 sec habituation period with guillotine doors closed.</li>
	<li>After habituation, open guillotine doors and allow mice free exploration of all 3 chambers for 1,200 sec, with time spent in each chamber being recorded using MedPC IV software (Med Associates, Inc.).</li>
	<li>Assign mice to conditioning chambers based on their performance during the pre-test. Using a biased design, pair drug administration during the subsequent 3 days with the compartment that was least-preferred during the baseline pre-test.</li>
	<li>During the next 3 conditioning days, inject mice with cocaine (10 mg/kg; i.p.) during the morning session and confine them in their assigned chamber for 1,200 sec immediately following the injection. Return mice to their home cages after the session. After 4 hr, inject mice with saline (0.01 ml/g body weight) and confine them to the opposite chamber for the 1,200 sec afternoon session.</li>
	<li>To test for acquisition, place mice in the central chamber with guillotine doors closed for a 60 sec habituation period. Open guillotine doors and allow free exploration for 1,200 sec, with time spent in all chambers recorded by MedPC IV software (Med Associates, Inc.). Calculate preference by subtracting the amount of time spent in the saline-paired chamber from the amount of time spent in the cocaine-paired chamber.</li>
	<li>To extinguish the place preference, expose mice to two 20 min sessions of free exploration daily in the same apparatus, separated by at least 2 hr, beginning 24 hr following the test for acquisition. Time spent in all chambers should again be recorded and the magnitude of the initial CPP should be compared with that of each extinction session.</li>
	<li>Determine whether mice have extinguished the drug-induced preference by assessing whether preference for a particular chamber across two consecutive extinction days is significantly lower compared to the acquisition preference score.</li>
	<li>Reinstate mice with a priming dose of cocaine (5 or 10 mg/kg; i.p.) and re-expose to the apparatus allowing free exploration and record the time spent in each compartment.</li>
	<li>Administer Euthasol euthanasia solution (150 mg/kg). Transcardially perfuse mice with 4% paraformaldehyde (PFA) and perform a histological examination to validate correct microinjection placement.</li>
</ol>

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The authors have nothing to disclose.

Regionally- and temporally-specific gene ablation via stereotaxic microinjection of viral vectors combined with CPP that includes extinction and reinstatement phases allows for investigation of the specific contributions of genes to three distinct phases of addictive-like behavior. While conditional knockout that is achieved utilizing the traditional Cre-LoxP system provides for spatio-temporally restricted gene ablation, stereotaxically microinjecting Cre-recombinase into discrete brain areas of floxed mice allows for e...

Cocaine is a highly reinforcing psychostimulant. Following repeated exposure, several molecular and cellular adaptations occur in reward-relevant brain circuitry that are believed to result in compulsive drug-seeking behavior, prompting high rates of relapse which pose a serious clinical problem 1. Cocaine exerts these long-lasting behavioral effects by regulating gene expression. To study the adaptations that arise from chronic cocaine use, preclinical rodent models have been used extensively. One such model is the conditioned place preference (CPP) paradigm. This model involves the development of a learned association between a previously neutral environment and the rewarding properties of cocaine. After several pairings of cocaine with a particular chamber, animals are allowed to freely explore the cocaine-paired and non-cocaine-paired environments and if they prefer the drug-paired compartment, they are said to have acquired a cocaine-induced place preference. Moreover, following an extinction training period, this paradigm can be used to study context-specific reinstatement of cocaine-seeking behavior.
Compared to other behavioral models of addictive-like behavior, such as psychomotor sensitization, which is used to study long-term cocaine-induced behavioral and molecular plasticity 2,3 and self-administration (SA), that is thought to more accurately mimic addictive-like behavior found in humans, the CPP paradigm is a simple procedure to study cocaine contextual learning 4. CPP protocols can be conveniently extended to include extinction and reinstatement phases, similarly to SA, which allow for investigation of the mechanisms underlying drug craving and relapse 5-7 and which are believed to recapitulate aspects of what occurs in human drug-seeking behavior and drug- and cue-induced relapse 8-10.
One mechanism that underlies the cocaine-induced behavioral plasticity that is characteristic of each of the distinct phases of CPP, including acquisition, extinction, and reinstatement, is activation of unique signatures of gene expression within different brain regions. To directly test which genes within reward-relevant brain regions mediate cocaine-induced behavioral changes, it is useful to be able to manipulate them selectively in a regionally-specific manner. One way of achieving this is to microinject recombinant adenoassociated viral (rAAV) vectors that express Cre recombinase using stereotaxic surgery, into distinct brain areas of mice that have target genes flanked by LoxP sites (floxed mice). This method allows for very precise temporal and regional control over when and where genes are ablated, in both dividing and non-dividing neurons, without inducing immune responses 11-13. This level of control represents an important advantage over traditional Cre-LoxP technology of breeding floxed mice with mice expressing Cre recombinase under the control of an endogenous gene promoter, in that the timing and distribution of gene deletion can be more tightly regulated. Additionally, viral vector-mediated gene knockout circumvents potential developmental compensatory effects that may occur using traditional knockout strategies.
In addition to cocaine CPP, which is described here, microinjection of rAAV-Cre into the brain of floxed mice to assess the role of specific genes can be applied ubiquitously to behavioral paradigms that involve separate phases, including self-administration and psychomotor sensitization. For example, our laboratory has utilized the rAAV-Cre technology to study the role of Cav1.2 L-type Ca2+ channels in cocaine psychomotor sensitization 14. Specifically, rAAV-Cre was microinjected into the nucleus accumbens (NAc) of mice with the gene encoding Cav1.2 floxed, to demonstrate that Cav1.2 acting in this region mediates the expression phase of psychomotor sensitization 14,15. However, the rAAV-Cre strategy cannot be used if mice with a floxed gene of interest do not exist, as was our experience when assessing the role of Cav1.3 L-type Ca2+ channels in psychomotor sensitization. Hence, a limitation of using rAAV-Cre presents itself if conditional mice do not exist for a particular gene of interest. However, rAAVs that express siRNA can be used to knockdown target genes, as we have done to examine the role of Cav1.3 channels 14,15.
Microinjecting rAAV-Cre into discrete brain regions of floxed mice and then testing them in the CPP paradigm allows for investigation into the specific genes that mediate the various phases of addictive-like behavior and where they are acting. Use of this paradigm has aided in our understanding of how repeated cocaine administration in essence hijacks the brains reward circuitry causing maladaptive changes in molecular signal transduction pathways and gene expression that lead to the addicted state 1,16,17.

<table><tbody><tr><td>Conditioned place preference activity chambers</td><td>Med Associates, Inc., St. Albans, VT, USA</td><td>MED-CPP-MS</td><td></td></tr><tr><td>Stereotaxic alignment system for mouse</td><td>David Kopf Instruments, Tujunga, CA, USA</td><td>model 900</td><td></td></tr><tr><td>Hamilton syringes</td><td>Hamilton Company, Reno, Nevada, USA</td><td>7634-01</td><td></td></tr><tr><td>rAAV2-Cre-GFP</td><td>Vector BioLabs, Philadelphia, PA, USA</td><td>7016</td><td></td></tr><tr><td>rAAV2-GFP</td><td>Vector BioLabs, Philadelphia, PA, USA</td><td>7004</td><td></td></tr></tbody></table>

stereotaxic microinjection of viral vectors expressing cre recombinase to study the role of target genes in cocaine conditioned place preference

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes of interest allows for enhanced temporally- and regionally-specific control of gene deletion, compared to existing methods. While conditional deletion can also be achieved by mating mice that express Cre recombinase under the control of specific gene promoters with mice carrying a floxed gene, stereotaxic microinjection allows for targeting of discrete brain areas at experimenter-determined time points of interest. In the context of cocaine conditioned place preference, and other cocaine behavioral paradigms such as self-administration or psychomotor sensitization that can involve withdrawal, extinction and/or reinstatement phases, this technique is particularly useful in exploring the unique contribution of target genes to these distinct phases of behavioral models of cocaine-induced plasticity. Specifically, this technique allows for selective ablation of target genes during discrete phases of a behavior to test their contribution to the behavior across time. Ultimately, this understanding allows for more targeted therapeutics that are best able to address the most potent risk factors that present themselves during each phase of addictive behavior.

CPP
After performing CPP on microinjected mice, one should verify that the cohort of control-injected (rAAV-GFP) mice have normally acquired preference for the drug-paired chamber (Figure 1A, 1B). Mice are considered to have acquired preference for a particular chamber when cocaine preference (time spent in the cocaine-paired chamber minus time spent in the saline-paired chamber) is significantly higher compared to baseline preference score (Figure 1B, A vs. B). ...

This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes ...

The overall goal of this procedure is to selectively ablate target genes during discrete phases of the condition place preference paradigm to test their contribution to this behavior across time. This is accomplished by first stereotaxic micro injecting the recombinant eno associated viral vector expressing Cree recombinase. Next, the mice are allowed to recover in their home cage for 10 to 14 days for maximal C reactivation.The third step of the procedure is to test the mice in conditioned place preference, including the acquisition extinction and reinstatement phases. The final step of the procedure is to perform a histological examination of micro injected mice to validate correct micro injection placement. Ultimately, results can be obtained that show changes in acquisition, extinction and or reinstatement behavior in response to gene knockout through condition place preference behavioral testing.The main advantage of this technique over existing methods like the CRE lock P mating system, is that it allows for more specific timing and anatomical specificity of the gene knockout discretely, allowing for the investigation of the loss of gene in each phase of the behavior. The implications of this technique extend towards treatment for addictive behavior because it involves better understanding the genetic basis that may underlie how exposure to contextual cues can trigger cocaine related memories that may prompt craving and relapse in addicts. To begin place a mouse cage with bedding on a heat lock to warm for postoperative recovery.Set up an electric razor for shaving the head and ethanol and iodine to sterilize the scalp. Use 70%ethanol to wipe down ear bars and mouth hold of the stereo attacks using a five microliter Hamilton syringe. Draw up five microliters of viral vector and set it up in a syringe holder On the stereo attacks.Place a microwaved heating pad or an electric pad on the stereo attacks after injecting the mouse with an IP injection of a ketamine xylazine cocktail. According to the text protocol, perform a tail pinch to check the level of sedation. Next, shave the head between the ears and eyes and use sterile cotton applicators to clean the area with alternating washes of ethanol and iodine.Then transfer the mouse to the heated pad on the stereo attacks zero. The tooth bar and ear bar scales and set the mouse's teeth in the tooth bar, screw in the muzzle and the ear bars. If any wiggling or whisker movement is noted.Administer a booster dose of ketamine xylazine cocktail. Then lubricate the eyes to prevent them from drying. Next, using a scalpel cut, open the scalp to just behind the ears.Place bulldog clips on the corners of the scalp to peel away the skin from the skull. Making sure Bgma and Lambda are easily visible. Ensure that the suture from bgma to Lambda is aligned and reposition the head if necessary.After confirming that the stereo attack settings are zeroed, put the needle tip in place at bma. Measure the dorsal ventral medial lateral and anterior posterior coordinates for both BMA and lambda. Then adjust the head until the ventral coordinates for BMA and lambda are within 0.02 millimeters of each other.Recenter the needle tip on bgma, and from that point moving in the anterior posterior direction to the desired coordinate. Next drill a borehole at the desired coordinate. Checking that the needle is able to enter through uninterrupted by the skull, then lower the needle to the desired ventral coordinate.Once the coordinate is reached, dip 0.015 millimeters below it for around 10 seconds to create a small pocket for the viral vector at a rate of roughly 0.1 microliters per minute, inject the desired volume of viral vector after waiting three minutes for the viral vector to diffuse, raise the needle tip 0.015 millimeters and wait an additional two minutes. When the injections are complete, apply bone wax to seal both holes and then suture the scalp. Finally, apply nerve block to the wounded area.Remove the mouse from the stereo attacks and place it in a warm cage on a heat block to recover for 45 minutes to an hour. Let the mouse recover for at least 10 days to allow for maximal reactivation. Give the animal water and soften food and buprenorphine twice daily if necessary for pain after recovery, place mice into the central chamber of a three chamber place preference apparatus for a 62nd habituation period with guillotine doors closed.After habituation, open the guillotine doors and allow the mice to freely explore all three chambers for 1200 seconds. Use Med PC four software to record the animal's time spent in each chamber using guidelines from the text protocol. Assign the mice to conditioning chambers based on their performance during the pretest.During the next three days, inject 10 milligrams per kilogram of cocaine during a morning session and immediately combine them in their assigned chamber for 1200 seconds. Return the mice to their home cages for four hours, then inject them with saline and immediately confine the mice to the opposite chamber for 1200 seconds. To test for acquisition, place the mice in the central chamber with guillotine doors closed for a 62nd habituation period.Open guillotine doors and allow free exploration for 1200 seconds or recording. Time spent in all chambers calculate preference by subtracting the amount of time spent in the saline pair chamber from the amount of time spent in the cocaine pair chamber. After performing CPP on micro injected mice, they're considered to have acquired preference for a particular chamber.When cocaine preference defined as time spent in the cocaine pair chamber minus time spent in the saline pair chamber is significantly higher compared to the baseline preference score. If as a cohort control injected mice do not show a preference for the cocaine pair chamber or show, an aversion placement is verified immediately via histology, and those mice with incorrect placement are eliminated from the study. If preference is normally acquired by the control injected cohort, the paradigm is extended by beginning extinction.Mice are considered to have extinguished the drug induced preference for a particular chamber. When cocaine preference across two consecutive extinction days is significantly lower compared to the acquisition preference score at this time, one can reinstate with a drug prime. If the preference score following administration of the drug prime is statistically indistinguishable from that at acquisition, then that cohort is said to have reinstated the drug-induced preference for the drug paired chamber using quantitative PCR.This figure demonstrates that the gene of interest has in fact been knocked out in micro injected mice following micro injection of RAAV Cree GFP into the nucleus accumbens that have the L type calcium channel isoform, CAV 1.2 flocks of RAAV. Cree injected punches show a significant decrease in gene expression compared to mice injected with A-A-V-G-F-P shown here. Immunohistochemical analysis of the nucleus accumbens for GFP reveals proper bilateral placement of micro injected RAAV Cree Following this procedure.Other techniques such as quantitative real-time PCR and Western bodying can be performed in order to answer questions regarding the effect of knockdown of a gene on the expression levels of other genes of interest. After Watching this video, you should have a better understanding of how to selectively ablate target genes in the mouse brain using stereotaxic microinjection viral vectors in order to selectively test the role of these genes in the acquisition extinction and reinstatement of the condition place preference paradigm.

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Source: Chen, S., et al., Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains. J. Vis. Exp. (2018)This video demonstrates the stereotaxic injection of a viral vector encoding Cre recombinase and green fluorescence protein or GFP into the striatum of a genetically modified mouse pup to achieve gene manipulation by excising a loxP-flanked STOP cassette, activating red fluorescence protein or RFP as a reporter.

Stereotactic Microinjection of a Viral Vector for Gene Manipulation in a Mouse Pup Striatum

Source: Pereira, M. et. al., In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors. J. Vis. Exp. (2019)This video demonstrates stereotaxic injection of an adeno-associated virus into the cerebral cortex of a transgenic mouse to deliver Cre-dependent neuronal reprogramming and fluorescent reporter genes into glial cells.

Stereotaxic Injection of Viral Vectors for Glial Cell Reprogramming in a Mouse Model

Source: Fricano-Kugler, C. J., et al. Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection. J. Vis. Exp. (2016)This video demonstrates stereotaxic viral injection for precise neuronal gene editing in the mouse brain. The protocol ensures accurate virus delivery to the dentate gyrus, enabling fluorescent labeling and CRISPR-mediated gene knockout to study neural circuits, gene function, and disease mechanisms in vivo.

Stereotaxic Viral Injection for Targeted Neuronal Gene Editing in the Mouse Brain

MicroRNAs have significant roles in brain structure and function. Here we describe a method to enforce hippocampal miRNA over-expression using stereotactic injection of an engineered miRNA-expressing lentivirus. This approach can serve as a relatively rapid way to assess the in vivo effects of over-expressed miRNAs in specific brain regions.

Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

Viral vectors allow for targeted gene manipulation. We demonstrate a method for conditional gene expression or ablation in the mouse spinal cord, using stereotaxic injection of a viral vector into the dorsal horn, a prominent site of synaptic contact between primary somatosensory afferents and neurons of the central nervous system.

Stereotaxic Injection of a Viral Vector for Conditional Gene Manipulation in the Mouse Spinal Cord

Stereotactic injection of lentiviruses expressing cDNAs or shRNAs can modulate gene expression in specific brain areas of mice. Here, we present a protocol to combine stereotactic injections with behavioral tasks, such as the Open Field Test (OFT) and the Forced Swim Test (FST).

Applying Stereotactic Injection Technique to Study Genetic Effects on Animal Behaviors

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Optogenetic approaches are widely used to manipulate neural activity and assess the consequences for brain function. Here, a technique is outlined that upon in vivo expression of the optical activator Channelrhodopsin, allows for ex vivo analysis of synaptic properties of specific long range and local neural connections in fear-related circuits.

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

The CRISPR/Cas9 system offers the potential to make targeted genome editing accessible and affordable to the scientific community. This protocol is intended to demonstrate how to create viruses that will knockout a gene of interest using the CRISPR/Cas9 system, and then inject them stereotaxically into the adult mouse brain.

Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection

We describe a protocol of stereotaxic surgery with a homemade head-fixed device for microinjecting reagents into the striatum of neonatal mouse brains. This technique allows genetic manipulation in neuronal cells of specific regions of neonatal mouse brains.

Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains

The methods described here outline a procedure used to optogenetically reverse cocaine-induced plasticity in a behaviorally-relevant circuit in rats. Sustained low-frequency optical stimulation of thalamo-amygdala synapses induces long-term depression (LTD). In vivo optogenetically-induced LTD in cocaine-experienced rats resulted in the subsequent attenuation of cue-motivated drug seeking.

Using Optogenetics to Reverse Neuroplasticity and Inhibit Cocaine Seeking in Rats

Learn stereotaxic microinjection of viral vectors expressing Cre recombinase to study gene function in cocaine conditioned place preference using mice. This method enables precise gene knockout during acquisition, extinction, and reinstatement phases.

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference (Video) | JoVE Journal

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference

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