Reelin demarcates a subset of pre‐Bötzinger complex neurons in adult rat

Identification of two markers of neurons in the pre‐Bötzinger complex (pre‐BötC), the neurokinin 1 receptor (NK1R) and somatostatin (Sst) peptide, has been of great utility in understanding the essential role of the pre‐BötC in breathing. Recently, the transcription factor dbx1 was identified as a critical, but transient, determinant of glutamatergic pre‐BötC neurons. Here, to identify additional markers, we constructed and screened a single‐cell subtractive cDNA library from pre‐BötC inspiratory neurons. We identified the glycoprotein reelin as a potentially useful marker, because it is expressed in distinct populations of pre‐BötC and inspiratory bulbospinal ventral respiratory group (ibsVRG) neurons. Reelin ibsVRG neurons were larger (27.1 ± 3.8 μm in diameter) and located more caudally (>12.8 mm caudal to Bregma) than reelin pre‐BötC neurons (15.5 ± 2.4 μm in diameter, <12.8 mm rostral to Bregma). Pre‐BötC reelin neurons coexpress NK1R and Sst. Reelin neurons were also found in the parahypoglossal and dorsal parafacial regions, pontine respiratory group, and ventromedial medulla. Reelin‐deficient (Reeler) mice exhibited impaired respones to hypoxia compared with littermate controls. We suggest that reelin is a useful molecular marker for pre‐BötC neurons in adult rodents and may play a functional role in pre‐BötC microcircuits. J. Comp. Neurol. 520:606–619, 2012. © 2011 Wiley Periodicals, Inc.

[1]  J. C. Smith,et al.  Pre-Bötzinger complex: a brainstem region that may generate respiratory rhythm in mammals. , 1991, Science.

[2]  T. Curran,et al.  Role of the reelin signaling pathway in central nervous system development. , 2001, Annual review of neuroscience.

[3]  P. Guyenet.  Neural structures that mediate sympathoexcitation during hypoxia. , 2000, Respiration physiology.

[4]  Yogesh K. Dwivedi,et al.  A decrease of reelin expression as a putative vulnerability factor in schizophrenia. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Joshua G Corbin,et al.  Developmental Origin of PreBötzinger Complex Respiratory Neurons , 2010, The Journal of Neuroscience.

[6]  L. Enquist,et al.  Innervation of the heart and its central medullary origin defined by viral tracing. , 1994, Science.

[7]  G. D’Arcangelo,et al.  The Reelin Signaling Pathway Promotes Dendritic Spine Development in Hippocampal Neurons , 2008, The Journal of Neuroscience.

[8]  E. Soriano,et al.  The Cells of Cajal-Retzius: Still a Mystery One Century After , 2005, Neuron.

[9]  Jun Ren,et al.  Fluorescent Tagging of Rhythmically Active Respiratory Neurons within the Pre-Bötzinger Complex of Rat Medullary Slice Preparations , 2005, The Journal of Neuroscience.

[10]  P. Fuller,et al.  Medullary Circuitry Regulating Rapid Eye Movement Sleep and Motor Atonia , 2009, The Journal of Neuroscience.

[11]  C. Walsh,et al.  Abnormal cerebellar development and axonal decussation due to mutations in AHI1 in Joubert syndrome , 2004, Nature Genetics.

[12]  N. Bunnett,et al.  Characterization of antibodies to the rat substance P (NK-1) receptor and to a chimeric substance P receptor expressed in mammalian cells , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[13]  A. Guidotti,et al.  Reelin is preferentially expressed in neurons synthesizing gamma-aminobutyric acid in cortex and hippocampus of adult rats. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Guyenet,et al.  Pre-Bötzinger neurons with preinspiratory discharges "in vivo" express NK1 receptors in the rat. , 2001, Journal of neurophysiology.

[15]  E. Costa,et al.  Epigenetic mechanisms expressed in basal ganglia GABAergic neurons differentiate schizophrenia from bipolar disorder , 2007, Schizophrenia Research.

[16]  I. Kohane,et al.  Gene regulation and DNA damage in the ageing human brain , 2004, Nature.

[17]  N. Mellen,et al.  Normal breathing requires preBötzinger complex neurokinin-1 receptor-expressing neurons , 2001, Nature Neuroscience.

[18]  A. Yamashita,et al.  Somatostatin and brain-derived neurotrophic factor mRNA expression in the primate brain: decreased levels of mRNAs during aging , 1997, Brain Research.

[19]  F. Panza,et al.  The RELN locus in Alzheimer's disease. , 2008, Journal of Alzheimer's disease : JAD.

[20]  E. Soriano,et al.  BDNF-modulated spatial organization of Cajal-Retzius and GABAergic neurons in the marginal zone plays a role in the development of cortical organization. , 2006, Cerebral cortex.

[21]  J. Siegel,et al.  Medullary regions mediating atonia , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[22]  M. Webster,et al.  Multivariate analysis of prefrontal cortical data from the Stanley Foundation Neuropathology Consortium , 2001, Brain Research Bulletin.

[23]  C. Sotelo,et al.  Regional and Cellular Patterns of reelin mRNA Expression in the Forebrain of the Developing and Adult Mouse , 1998, The Journal of Neuroscience.

[24]  J C Smith,et al.  Pre-Bötzinger complex in the cat. , 1995, Journal of neurophysiology.

[25]  Daniel Choquet,et al.  NMDA Receptor Surface Trafficking and Synaptic Subunit Composition Are Developmentally Regulated by the Extracellular Matrix Protein Reelin , 2007, The Journal of Neuroscience.

[26]  Yogesh K. Dwivedi,et al.  Decrease in reelin and glutamic acid decarboxylase67 (GAD67) expression in schizophrenia and bipolar disorder: a postmortem brain study. , 2000, Archives of general psychiatry.

[27]  E. Callaway,et al.  Silencing preBötzinger Complex somatostatin-expressing neurons induces persistent apnea in awake rat , 2008, Nature Neuroscience.

[28]  Jonathan A. Cooper.  A mechanism for inside-out lamination in the neocortex , 2008, Trends in Neurosciences.

[29]  P. Sawchenko,et al.  Organization and Transmitter Specificity of Medullary Neurons Activated by Sustained Hypertension: Implications for Understanding Baroreceptor Reflex Circuitry , 1998, The Journal of Neuroscience.

[30]  H. Kayserili,et al.  Mutations in the AHI1 gene, encoding jouberin, cause Joubert syndrome with cortical polymicrogyria. , 2004, American journal of human genetics.

[31]  M. Hamburgh.  ANALYSIS OF THE POSTNATAL DEVELOPMENTAL EFFECTS OF "REELER," A NEUROLOGICAL MUTATION IN MICE. A STUDY IN DEVELOPMENTAL GENETICS. , 1963, Developmental biology.

[32]  J. Feldman,et al.  Projections of preBötzinger Complex neurons in adult rats , 2010, The Journal of comparative neurology.

[33]  J. Feldman,et al.  Modulation of respiratory frequency by peptidergic input to rhythmogenic neurons in the preBötzinger complex. , 1999, Science.

[34]  J. Rihel,et al.  Single-Cell Transcriptional Analysis of Neuronal Progenitors , 2003, Neuron.

[35]  Johan Ericson,et al.  Hindbrain interneurons and axon guidance signaling critical for breathing , 2010, Nature Neuroscience.

[36]  J A Neubauer,et al.  Pre-Bötzinger complex functions as a central hypoxia chemosensor for respiration in vivo. , 2000, Journal of neurophysiology.

[37]  D. Falconer.  Two new mutants, ‘trembler’ and ‘reeler’, with neurological actions in the house mouse (Mus musculus L.) , 2008, Journal of Genetics.

[38]  T. Curran,et al.  A protein related to extracellular matrix proteins deleted in the mouse mutant reeler , 1995, Nature.

[39]  J. Champagnat,et al.  Central control of breathing in mammals: neuronal circuitry, membrane properties, and neurotransmitters. , 1995, Physiological reviews.

[40]  G. D’Arcangelo.  The role of the Reelin pathway in cortical development. , 2000, Symposia of the Society for Experimental Biology.

[41]  J. Herz,et al.  Reelin, lipoprotein receptors and synaptic plasticity , 2006, Nature Reviews Neuroscience.

[42]  K. Deisseroth,et al.  Active Expiration Induced by Excitation of Ventral Medulla in Adult Anesthetized Rats , 2011, The Journal of Neuroscience.

[43]  Isolation of an allele of reeler by insertional mutagenesis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Hong Wang,et al.  A group of glutamatergic interneurons expressing high levels of both neurokinin‐1 receptors and somatostatin identifies the region of the pre‐Bötzinger complex , 2003, The Journal of comparative neurology.

[45]  M. Frotscher,et al.  Modulation of Synaptic Plasticity and Memory by Reelin Involves Differential Splicing of the Lipoprotein Receptor Apoer2 , 2005, Neuron.

[46]  Eduardo Soriano,et al.  Reelin Regulates Postnatal Neurogenesis and Enhances Spine Hypertrophy and Long-Term Potentiation , 2010, The Journal of Neuroscience.

[47]  M. Chesselet,et al.  Preprotachykinin messenger RNA detected by in situ hybridization in striatal neurons of the human brain , 1987, Brain Research.

[48]  C. Quattrocchi,et al.  Reelin Promotes Hippocampal Dendrite Development through the VLDLR/ApoER2-Dab1 Pathway , 2004, Neuron.

[49]  A. Guidotti,et al.  Cortical bitufted, horizontal, and Martinotti cells preferentially express and secrete reelin into perineuronal nets, nonsynaptically modulating gene expression. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[50]  J. D. del Río,et al.  Reelin expression and glycosylation patterns are altered in Alzheimer's disease. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[51]  N. Mehta,et al.  Ectopic expression of reelin alters migration of sympathetic preganglionic neurons in the spinal cord , 2009, The Journal of comparative neurology.

[52]  C. Blakemore,et al.  The Role of the First Postmitotic Cortical Cells in the Development of Thalamocortical Innervation in the ReelerMouse , 1998, The Journal of Neuroscience.

[53]  P. Mantyh,et al.  Inhibition of hyperalgesia by ablation of lamina I spinal neurons expressing the substance P receptor. , 1997, Science.