Optogenetic Functional Magnetic Resonance Imaging (OfMRI): Genetically Targeted in Vivo Brain Circuit Mapping
Jin Hyung Lee¹, Remy Durand², Viviana Gradinaru², Feng Zhang², Dae-Shik Kim³, Karl Deisseroth^2
1Electrical Engineering, University of California, Los Angeles, Los Angeles, CA, United States; ²Bioengineering, Stanford University, Stanford, CA, United States; ³Boston University, Boston, MA, United States

Despite an enormous, rapidly-growing functional brain imaging literature based on blood oxygenation level dependent (BOLD) signals, it remains controversial which classes of local activity and cellular elements (e.g., glia, axonal tracts, or excitatory neurons) can trigger BOLD responses. Using a novel methodology integrating Optogenetics with high-field fMRI, we show here that robust BOLD signal can be triggered in primary motor cortex by specific recruitment of CaMKIIa-expressing excitatory neurons. We further show that this approach allows for highly specific in vivo circuit identification, in which the functional role of cell types defined by location and genetic identity, can be directly observed and globally mapped in the living mammal.

Light-Induced Activation of Light-Sensitive Pumps Modulates FMRI Responses - not available
John E. Downey^1,2, Piotr Walczak^3,4, Suresh E. Joel^1,2, Assaf A. Gilad^3,4, Michael T. McMahon^1,2, Heechul Kim^3,4, James J. Pekar^1,2, Galit Pelled^2,5
1F.M. Kirby Research Center for Functional Brain Imaging, Kennedy Krieger Institute, Baltimore, MD, United States; ²The Russell H. Morgan Department of Radiology and Radiological Sciences , Johns Hopkins University School of Medicine, Baltimore, MD, United States; ³The Russell H. Morgan Department of Radiology and Radiological Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, United States; ⁴Cellular Imaging Section, Vascular Biology Program, Institute for Cell Engineering , Johns Hopkins University School of Medicine, Baltimore, MD, United States; ⁵F.M. Kirby Research Center for Functional Brain Imaging, Kennedy Krieger Institute, Baltimore, MD, United States

Recent developments in optical-genetic (optogenetics) approaches enable immediate manipulations of neuronal firing rate by using light-induced activation of light sensitive pumps. We have engineered the excitatory neurons in rat somatosensory cortex to express halorhodopsin (light-sensitive chloride pump) using direct neuronal infection with lentivirus. Thus, in the presence of light, the chloride pumps open and trigger neuronal hyperpolarization i.e. decreases in neuronal firing rate. Consistent with electrophysiology results, light induced activation of halorhodopsin during forepaw stimulation, decreased the amplitude and the extent of fMRI responses. These results introduce an exciting and novel approach to study neuronal behavior in vivo.

In-Vivo Optogenetic Activation of Cortical Astrocytes with FMRI at 9.4T: OptoMRI
Jack A. Wells¹, Simon Walker-Samuel¹, Nephtali Marina², Melina Figueiredo³, Anja G. Teschemacher³, Michael Spyer², Alexander V. Gourine², Sergey Kasparov³, Mark F. Lythgoe^1
1Centre for Advanced Biomedical Imaging, University College London, London, United Kingdom; ²Neuroscience, Physiology & Pharmacology, University College London, London, United Kingdom; ³Physiology & Pharmacology, University of Bristol, Bristol, United Kingdom

The relative contribution of the neuronal and glial activation to the BOLD signals is not fully established. Optogenetic techniques, in which particular brain cells are engineered to express light-sensitive ion channels, offer minimally invasive and temporally precise control of the activities of distinct cellular populations.
In this study we performed simultaneous optogenetic activation of cortical astrocytes with high field fMRI . Astrocytes in the cortex of the anaesthetised rat brain were stimulated during continuous imaging using gradient echo EPI at 9.4T.  Here we present our preliminary data.

Mapping the Circuit of Fear with Pharmacogenetic Silencing and FMRI
Alessandro Gozzi¹, Apar Jain², Valerio Crestan¹, Adam J. Schwarz^1,3, Theodoros Tsetsenis², Graham Sheridan⁴, Cornelius T. Gross⁴, Angelo Bifone^1
1Neuroscience CEDD, GlaxoSmithKline, Verona, Verona, Italy; ²Mouse Biology Unit, EMBL, , Monterotondo, Italy; ³Translational Imaging, Eli Lilly, Indianapolis, IN, United States; ⁴Mouse Biology Unit, EMBL,, Monterotondo, Italy

Functional MRI methods have been widely applied to map regional changes in brain activity elicited by somatosensory stimuli, complex cognitive or emotional tasks, and pharmacological challenges. Here we describe and demonstrate the use of fMRI to map the functional effects of rapid and reversible pharmacogenetic silencing of selected neuronal populations focally expressed in specific regions of the mouse brain. In combination with behavioural observations, this novel approach provides a powerful means to assess the functional role of these neurons, to resolve the brain circuitry they are elements of, and to establish their implication in behavioural control.

Pharmacological MRI and Resting-State FMRI of Functional Brain Organization in the Serotonin Transporter Knock-Out Rat
Kajo van der Marel¹, Judith R. Homberg², Willem M. Otte¹, Rick M. Dijkhuizen^1
1Image Sciences Institute, University Medical Center Utrecht, Utrecht, Netherlands; ²Donders Centre for Neuroscience, UMC St. Radboud, Nijmegen, Netherlands

Genetic variation in the serotonin transporter gene (5-HTTLPR) has been linked to various neuropsychiatric disorders, including depression and drug addiction. In this study we combined resting-state fMRI (rs-fMRI) with pharmacological fMRI (phMRI) in the serotonin transporter knock-out rat, to study the effects of disrupted serotonin homeostasis on functional organization during baseline and psychoactive stimulation. With rs-fMRI we observed positive functional connectivity among ROIs within the limbic system, but no difference with controls. With phMRI we found stronger activation responses to cocaine in knock-outs in specific limbic areas, which is in agreement with previously reported cocaine supersensitivity.

Differential Effects of Chronic Fluoxetine Use in Young Vs. Adult Rats: A PhMRI Study
Anne Klomp¹, Jordi L. Tremoleda², Aart J. Nederveen¹, Marzena Wylezinska², Willy Gsell², Liesbeth Reneman^1
1Department of Radiology, Academical Medical Center, Amsterdam, Netherlands; ²Biological Imaging Centre, Imaging Science Department, MRC Clinical Sciences Centre, Hammersmith Hospital, Imperial College London, London, United Kingdom

The effects of chronic fluoxetine treatment (the only SSRI registered for use in children) on the developing brain are not well studied. Here we investigate the effect of chronic fluoxetine exposure on the serotonergic system in adult and peri-adolescent rats using phMRI. Chronic treatment with fluoxetine elicits a reduction of overall brain activation in adult rats but not in young rats. Previous data from our group showed an increase of serotonin transporters after chronic treatment in peri-adolescent rats but not in adult rats, suggesting a compensation mechanism occurring in the developing brain which could explain our phMRI findings.

Endogenous Opioid-Dopamine Neurotransmission Evokes Sustained Negative CBV-Weighted FMRI Responses
Yen-Yu Ian Shih^1,2, Yun-Chen Chiang^2,3, Yi-Hua Hsu², Fu-Shan Jaw³, Jin-Chung Chen⁴, Bai-Chuang Shyu², Timothy Q. Duong¹, Chen Chang^2
1Research Imaging Institute, University of Texas Health Science Center at San Antonio, San Antonio, TX, United States; ²Functional and Micro-Magnetic Resonance Imaging Center, Academia Sinica, Taipei, Taiwan; ³Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan; ⁴Department of Physiology and Pharmacology, Chang Gung University, Taoyuan, Taiwan

Dopamine and opioids have been implicated in various aspects of brain signaling. By employing CBV-weighted fMRI with pharmacological treatments, the present study reveals that endogenous stimulation of ?opioid receptors underlies negative CBV fMRI signals via the activation of dopamine D2/D3 receptors. The interpretation of fMRI data involving opioid–dopamine interactions requires careful consideration.

Differential Effect of Adrenoceptor on Functional Activation and Connectivity
Fatima Ali Nasrallah¹, Jolena Tan, Nora Hennies, Kai-Hsiang Chuang
¹Lab of Molecular Imaging, Singapore Bioimaging Consortium, Singapore, Singapore

In this work we clearly demonstrate the modulation of resting state functional connectivity by the á2-adrenergic receptor agonist, medetomidine. We determined the functional activation response induced by forepaw stimulation under 0.1, 0.2, and 0.3 mg/kg/hr infusion of medetomidine and the corresponding resting state functional connectivity as well. While BOLD signal change was unchanged across dosages, medetomidine had a profound effect on the synchronicity of interacting regions in the brain.

Anaesthetic Interactions in the PhMRI Response to Acute Ketamine Challenge
Duncan Jack Hodkinson¹, Carmen de Groote², Shane McKie³, John-Francis William Deakin³, Steve R. Williams^1
1Imaging Science and Biomedical Engineering, University of Manchester, Manchester, United Kingdom; ²Neuroscience and Biomedical Systems, University of Glasgow, Glasgow, United Kingdom; ³Neuroscience and Psychiatry Unit, University of Manchester, Manchester, United Kingdom

Pharmacological-challenge MRI (phMRI) is an exciting new tool enabling researchers to examine underlying circuitry of the brain in response to neuroactive drugs. To avoid head movements pre-clinical phMRI studies are often conducted under general anaesthesia. However, interactions between the drug of interest and the anaesthetic may be a confounding factor. Here we assessed the effect of α-chloralose and isoflurane anesthesia on the phMRI response to ketamine challenge. The positive BOLD signal changes observed with α-chloralose showed areas of activation similar to neuroimaging studies in humans. A drug-anaesthetic interaction between isoflurane and ketamine compromised the phMRI response.

Simultaneous FMRI and Local Field Potential Measurements of Epileptic Seizures in Medetomidine Sedated and Awake Rats
Antti Markku Airaksinen¹, Shahryar Khan Hekmatyar², Neil Jerome², Juha-Pekka Niskanen^1,3, Asla Pitkanen^4,5, Risto A. Kauppinen², Olli Grohn^1
1Department of Neurobiology, A.I.Virtanen institute for Molecular Sciences, University of Kuopio, Kuopio, Finland; ²Dartmouth Medical School, Biomedical NMR Research Center, Hanover, NH, United States; ³Department of Physics, University of Kuopio, Kuopio, Finland; ⁴Department of Neurobiology, Epilepsy Research Laboratory, A.I.Virtanen institute, University of Kuopio, Kuopio, Finland; ⁵Department of Neurology, Kuopio University Hospital, Kuopio, Finland

Simultaneous LFP and fMRI measurements were performed during kainic acid (KA) induced seizures in awake and medetomidine anesthetized rats. The recurrent epileptic seizures were detected in the LFP signal after KA injection and robust BOLD responses were observed in the hippocampus both in awake and sedated animals.  To determine basal CBF, ASL was performed showing the highest CBF values in isoflurane anesthetized rats and the lowest CBF under medetomidine sedation. We conclude that medetomidine sedation is suitable for studies of normal and abnormal brain activity, but lowered basal CBF level should be taken into account when interpreting the fMRI results.