A reliable protocol for the manual segmentation of the human amygdala and its subregions using ultra-high resolution MRI

The measurement of the volume of the human amygdala in vivo has received increasing attention over the past decade, but existing methods face several challenges. First, due to the amorphous appearance of the amygdala and the difficulties in interpreting its boundaries, it is common for protocols to omit sizable sections of the rostral and dorsal regions of the amygdala comprising parts of the basolateral complex (BL) and central nucleus (Ce), respectively. Second, segmentation of the amgydaloid complex into separate subdivisions is challenging due to the resolution of routinely acquired images and the lack of standard protocols. Recent advances in technology have made ultra-high resolution MR images available, and in this study we provide a detailed segmentation protocol for manually tracing the whole amygdala that incorporates a greater portion of the rostral and dorsal sections with techniques illustrated in detail to maximize reproducibility. In addition, we propose a geometrically-based protocol for segmenting the amygdala into four component subregions of interest (sROI), which correspond largely to amygdala subnuclear divisions: the BL sROI, centromedial (CM) sROI, basomedial (BM) sROI, and the amygdaloid cortical (ACo) sROI. We performed an intra- and inter-rater reliability study of our methods in 10 adults (5 young adults and 5 older adults). The results indicate that both protocols can be implemented with a high degree of reliability (the majority of intra-rater and inter-rater correlations were > 0.81). This protocol should aid further research into the alterations in amygdala anatomy, connectivity, and function that accompany normal aging and pathology associated with neuropsychiatric disorders.

[1]  S. Rauch,et al.  Masked Presentations of Emotional Facial Expressions Modulate Amygdala Activity without Explicit Knowledge , 1998, The Journal of Neuroscience.

[2]  D. Amaral,et al.  The Amygdala, Social Behavior, and Danger Detection , 2003, Annals of the New York Academy of Sciences.

[3]  Koenraad Van Leemput,et al.  Model-Based Segmentation of Hippocampal Subfields in Ultra-High Resolution In Vivo MRI , 2008, MICCAI.

[4]  H. Albers,et al.  Role of the central amygdala in social communication in Syrian hamsters (Mesocricetus auratus) , 1997, Brain Research.

[5]  Leslie G. Ungerleider,et al.  Neural processing of emotional faces requires attention , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Daniel S. Margulies,et al.  Functional connectivity of the human amygdala using resting state fMRI , 2009, NeuroImage.

[7]  L. R. Dice.  Measures of the Amount of Ecologic Association Between Species , 1945 .

[8]  F Andermann,et al.  Anatomic basis of amygdaloid and hippocampal volume measurement by magnetic resonance imaging , 1992, Neurology.

[9]  Martin Styner,et al.  A comparison of automated segmentation and manual tracing for quantifying hippocampal and amygdala volumes , 2009, NeuroImage.

[10]  Lawrence L. Wald,et al.  A 7 Tesla Gradient Mode Birdcage Coil for Improved Temporal and Occipital Lobe SNR , 2006 .

[11]  R. Palmiter,et al.  Long-Term Memory for Pavlovian Fear Conditioning Requires Dopamine in the Nucleus Accumbens and Basolateral Amygdala , 2010, PloS one.

[12]  Angela D. Friederici,et al.  Diffusion tensor imaging segments the human amygdala in vivo , 2010, NeuroImage.

[13]  Mark D'Esposito,et al.  Behavioral / Systems / Cognitive Amygdala Response to Facial Expressions Reflects Emotional Learning , 2006 .

[14]  J. D. McGaugh,et al.  Amygdala modulation of hippocampal-dependent and caudate nucleus-dependent memory processes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Anders M. Dale,et al.  Automated manifold surgery: constructing geometrically accurate and topologically correct models of the human cerebral cortex , 2001, IEEE Transactions on Medical Imaging.

[16]  Yutaka Matsuoka,et al.  [Manual tracing guideline for volumetry of hippocampus and amygdala with high-resolution MRI]. , 2003, No to shinkei = Brain and nerve.

[17]  L. Bourgeais,et al.  Projections from the nociceptive area of the central nucleus of the amygdala to the forebrain: a PHA‐L study in the rat , 2001, The European journal of neuroscience.

[18]  R. Adolphs,et al.  Impaired recognition of emotion in facial expressions following bilateral damage to the human amygdala , 1994, Nature.

[19]  J. Fleiss,et al.  Intraclass correlations: uses in assessing rater reliability. , 1979, Psychological bulletin.

[20]  Joseph E LeDoux,et al.  Different projections of the central amygdaloid nucleus mediate autonomic and behavioral correlates of conditioned fear , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[21]  Gabriele Lohmann,et al.  Parcellation of human amygdala in vivo using ultra high field structural MRI , 2011, NeuroImage.

[22]  M. Zarrindast,et al.  Morphine-induced place preference: Involvement of the central amygdala NMDA receptors , 2007, Brain Research.

[23]  Martin Styner,et al.  Rebuttal to Hasan and Pedraza in comments and controversies: “Improving the reliability of manual and automated methods for hippocampal and amygdala volume measurements” , 2009, NeuroImage.

[24]  P. Holland,et al.  Effects of reward timing information on cue associability are mediated by amygdala central nucleus. , 2011, Behavioral neuroscience.

[25]  S. Rauch,et al.  A functional MRI study of human amygdala responses to facial expressions of fear versus anger. , 2001, Emotion.

[26]  Polina Golland,et al.  Automated segmentation of hippocampal subfields from ultra‐high resolution in vivo MRI , 2009, Hippocampus.

[27]  Anders M. Dale,et al.  On-line automatic slice positioning for brain MR imaging , 2005, NeuroImage.

[28]  Amit Anand,et al.  Brain Imaging Studies in Mood and Anxiety Disorders , 2003, Annals of the New York Academy of Sciences.

[29]  Michael W. Miller,et al.  Cortical connections between rat cingulate cortex and visual, motor, and postsubicular cortices , 1983, The Journal of comparative neurology.

[30]  E. Yeterian,et al.  MRI-Based Topographic Parcellation of Human Cerebral White Matter and Nuclei II. Rationale and Applications with Systematics of Cerebral Connectivity , 1999, NeuroImage.

[31]  Michael A Yassa,et al.  A quantitative evaluation of cross-participant registration techniques for MRI studies of the medial temporal lobe , 2009, NeuroImage.

[32]  J M Besson,et al.  Nucleus centralis of the amygdala and the globus pallidus ventralis: electrophysiological evidence for an involvement in pain processes. , 1992, Journal of neurophysiology.

[33]  Anders M. Dale,et al.  Cortical Surface-Based Analysis I. Segmentation and Surface Reconstruction , 1999, NeuroImage.

[34]  André J. W. van der Kouwe,et al.  Brain morphometry with multiecho MPRAGE , 2008, NeuroImage.

[35]  A. Dale,et al.  Whole Brain Segmentation Automated Labeling of Neuroanatomical Structures in the Human Brain , 2002, Neuron.

[36]  Anders M. Dale,et al.  An automated labeling system for subdividing the human cerebral cortex on MRI scans into gyral based regions of interest , 2006, NeuroImage.

[37]  A M Dale,et al.  Measuring the thickness of the human cerebral cortex from magnetic resonance images. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[38]  K. Amunts,et al.  Cytoarchitectonic mapping of the human amygdala, hippocampal region and entorhinal cortex: intersubject variability and probability maps , 2005, Anatomy and Embryology.

[39]  Alan C. Evans,et al.  Volumetry of hippocampus and amygdala with high-resolution MRI and three-dimensional analysis software: minimizing the discrepancies between laboratories. , 2000, Cerebral cortex.

[40]  Robert Zivadinov,et al.  Abnormal subcortical deep-gray matter susceptibility-weighted imaging filtered phase measurements in patients with multiple sclerosis A case-control study , 2012, NeuroImage.

[41]  P. Holland,et al.  Dissociable effects of disconnecting amygdala central nucleus from the ventral tegmental area or substantia nigra on learned orienting and incentive motivation , 2007, The European journal of neuroscience.

[42]  A. Marcilhac,et al.  Identification of projections from the central nucleus of the amygdala to the paraventricular nucleus of the hypothalamus which are immunoreactive for corticotrophin‐releasing hormone in the rat , 1997, Experimental physiology.

[43]  L. Heimer,et al.  The Concepts of the Ventral Striatopallidal System and Extended Amygdala , 1999, Annals of the New York Academy of Sciences.

[44]  Alexander Hammers,et al.  Fully Automatic Segmentation of the Hippocampus and the Amygdala from MRI Using Hybrid Prior Knowledge , 2007, MICCAI.

[45]  Michael Davis,et al.  The amygdala , 2000, Current Biology.

[46]  J. Price.  Prefrontal Cortical Networks Related to Visceral Function and Mood , 1999, Annals of the New York Academy of Sciences.

[47]  R. Dolan,et al.  Conscious and unconscious emotional learning in the human amygdala , 1998, Nature.

[48]  G. Paxinos,et al.  THE HUMAN NERVOUS SYSTEM , 1975 .

[49]  D. Gaffan,et al.  Visual-visual associative learning and reward-association learning in monkeys: the role of the amygdala , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[50]  L. Medina,et al.  Distribution of nitric oxide-producing neurons in the developing and adult mouse amygdalar basolateral complex , 2005, Brain Research Bulletin.

[51]  H. Ursin,et al.  KAADA BR: Subcortical structures mediating the attention response induced by amygdala stimulation. , 1960, Experimental neurology.

[52]  Anders M. Dale,et al.  A hybrid approach to the Skull Stripping problem in MRI , 2001, NeuroImage.

[53]  J. Polimeni,et al.  96‐Channel receive‐only head coil for 3 Tesla: Design optimization and evaluation , 2009, Magnetic resonance in medicine.

[54]  M. Haine,et al.  Van Damme A. , 1986 .

[55]  A. Dale,et al.  Cortical Surface-Based Analysis II: Inflation, Flattening, and a Surface-Based Coordinate System , 1999, NeuroImage.

[56]  Joseph E LeDoux,et al.  Contributions of the Amygdala to Emotion Processing: From Animal Models to Human Behavior , 2005, Neuron.

[57]  H. Barbas.  Connections underlying the synthesis of cognition, memory, and emotion in primate prefrontal cortices , 2000, Brain Research Bulletin.

[58]  E. Phelps,et al.  The human amygdala. , 2009 .

[59]  Leslie G. Ungerleider,et al.  Attentional control of the processing of neural and emotional stimuli. , 2002, Brain research. Cognitive brain research.

[60]  T. Kishi,et al.  Topographical projection from the hippocampal formation to the amygdala: A combined anterograde and retrograde tracing study in the rat , 2006, The Journal of comparative neurology.

[61]  Leonardo Bonilha,et al.  Protocol for volumetric segmentation of medial temporal structures using high‐resolution 3‐D magnetic resonance imaging , 2004, Human brain mapping.

[62]  H E ROSVOLD,et al.  Influence of amygdalectomy on social behavior in monkeys. , 1954, Journal of comparative and physiological psychology.

[63]  Balaji Krishnan,et al.  Long-term potentiation (LTP) in the central amygdala (CeA) is enhanced after prolonged withdrawal from chronic cocaine and requires CRF1 receptors. , 2007, Journal of neurophysiology.

[64]  Fabrice Bartolomei,et al.  Emotion induction after direct intracerebral stimulations of human amygdala. , 2007, Cerebral cortex.

[65]  Lisa Feldman Barrett,et al.  The amygdala and the experience of affect. , 2007, Social cognitive and affective neuroscience.

[66]  Nathan J Emery,et al.  Bilateral neurotoxic amygdala lesions in rhesus monkeys (Macaca mulatta): consistent pattern of behavior across different social contexts. , 2008, Behavioral neuroscience.

[67]  A. Dale,et al.  High‐resolution intersubject averaging and a coordinate system for the cortical surface , 1999, Human brain mapping.

[68]  P. Holland,et al.  Amygdala circuitry in attentional and representational processes , 1999, Trends in Cognitive Sciences.

[69]  K. Hsu,et al.  Differential roles of basolateral and central amygdala on the effects of uncontrollable stress on hippocampal synaptic plasticity , 2008, Hippocampus.

[70]  Karl H. Pribram,et al.  Influence of amygdalectomy on social behavior in monkeys. , 1954, Journal of comparative and physiological psychology.

[71]  Oliver T Wolf,et al.  MRI volume of the amygdala: a reliable method allowing separation from the hippocampal formation , 1999, Psychiatry Research: Neuroimaging.

[72]  Eric Achten,et al.  Intra- and interobserver variability of MRI-based volume measurements of the hippocampus and amygdala using the manual ray-tracing method , 1998, Neuroradiology.

[73]  C. Grillon,et al.  Phasic vs Sustained Fear in Rats and Humans: Role of the Extended Amygdala in Fear vs Anxiety , 2010, Neuropsychopharmacology.

[74]  D. Pandya,et al.  Cingulate cortex of the rhesus monkey: II. Cortical afferents , 1987, The Journal of comparative neurology.

[75]  E. Phelps,et al.  Neural mechanisms mediating optimism bias , 2007, Nature.

[76]  G. Paxinos,et al.  Atlas of the Human Brain , 2000 .

[77]  Bruce Fischl,et al.  Connectivity-based segmentation of human amygdala nuclei using probabilistic tractography , 2011, NeuroImage.

[78]  Dominique Hasboun,et al.  Anatomically constrained region deformation for the automated segmentation of the hippocampus and the amygdala: Method and validation on controls and patients with Alzheimer’s disease , 2007, NeuroImage.

[79]  J. R. Landis,et al.  The measurement of observer agreement for categorical data. , 1977, Biometrics.

[80]  D. Louis Collins,et al.  Appearance-based modeling for segmentation of hippocampus and amygdala using multi-contrast MR imaging , 2011, NeuroImage.

[81]  D. Coon.  The Human Nervous System 2nd ed , 1975 .

[82]  Timothy E. J. Behrens,et al.  Deep and Superficial Amygdala Nuclei Projections Revealed In Vivo by Probabilistic Tractography , 2011, The Journal of Neuroscience.

[83]  Khader M. Hasan,et al.  Improving the reliability of manual and automated methods for hippocampal and amygdala volume measurements , 2009, NeuroImage.

[84]  H Scheich,et al.  Auditory perception of laughing and crying activates human amygdala regardless of attentional state. , 2001, Brain research. Cognitive brain research.

[85]  D. Amaral,et al.  An autoradiographic study of the projections of the central nucleus of the monkey amygdala , 1981, The Journal of neuroscience : the official journal of the Society for Neuroscience.