Sex Differences in the Cerebellum and Frontal Cortex: Roles of Estrogen Receptor Alpha and Sex Chromosome Genes

Most neurobehavioral diseases are sexually dimorphic in their incidence, and sex differences in the brain may be key for understanding and treating these diseases. Calbindin (Calb) D28K is used as a biomarker for the well-studied sexually dimorphic nucleus, a hypothalamic structure that is larger in males than in females. In the current study weanling C56BL/6J mice were used to examine sex differences in the Calb protein and message focusing on regions outside of the hypothalamus. A robust sex difference was found in Calb in the frontal cortex (FC) and cerebellum (CB; specifically in Purkinje cells); mRNA and protein were higher in females than in males. Using 2 mouse lines, i.e. one with a complete deletion of estrogen receptor alpha (ERα) and the other with uncoupled gonads and sex chromosomes, we probed the mechanisms that underlie sexual dimorphisms. In the FC, deletion of ERα reduced Calb1 mRNA in females compared to males. In addition, females with XY sex chromosomes had levels of Calb1 equal to those of males. Thus, both ERα and the sex chromosome complement regulate Calb1 in the FC. In the CB, ERα knockout mice of both sexes had reduced Calb1 mRNA, yet sex differences were retained. However, the sex chromosome complement, regardless of gonadal sex, dictated Calb1 mRNA levels. Mice with XX chromosomes had significantly greater Calb1 than did XY mice. This is the first study demonstrating that sex chromosome genes are a driving force producing sex differences in the CB and FC, which are neuoranatomical regions involved in many normal functions and in neurobehavioral diseases.

[1]  D. Geschwind,et al.  Connecting genes to brain in the autism spectrum disorders. , 2010, Archives of neurology.

[2]  M. Niculescu,et al.  Choline deficiency alters global histone methylation and epigenetic marking at the Rel site of the calbindin 1 gene , 2010, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[3]  Henning Tiemeier,et al.  Cerebellum development during childhood and adolescence: A longitudinal morphometric MRI study , 2010, NeuroImage.

[4]  K. Merikangas,et al.  Prevalence and Treatment of Mental Disorders Among US Children in the 2001–2004 NHANES , 2010, Pediatrics.

[5]  J. Gray,et al.  Variation in orbitofrontal cortex volume: relation to sex, emotion regulation and affect. , 2009, Social cognitive and affective neuroscience.

[6]  P. Strata,et al.  Interactions between neuroactive steroids and reelin haploinsufficiency in Purkinje cell survival , 2009, Neurobiology of Disease.

[7]  S. Nakanishi,et al.  Genetic manipulation study of information processing in the cerebellum , 2009, Neuroscience.

[8]  T. Shors,et al.  Sex differences in learning processes of classical and operant conditioning , 2009, Physiology & Behavior.

[9]  M. McCarthy,et al.  New tricks by an old dogma: Mechanisms of the Organizational / Activational Hypothesis of steroid-mediated sexual differentiation of brain and behavior , 2009, Hormones and Behavior.

[10]  D. Hamson,et al.  Analysis of Foxp2 expression in the cerebellum reveals a possible sex difference , 2009, Neuroreport.

[11]  A. Arnold,et al.  Mouse Models for Evaluating Sex Chromosome Effects that Cause Sex Differences in Non‐Gonadal Tissues , 2009, Journal of neuroendocrinology.

[12]  Joseph M. Andreano,et al.  Sex influences on the neurobiology of learning and memory. , 2009, Learning & memory.

[13]  A. Arnold,et al.  X chromosome number causes sex differences in gene expression in adult mouse striatum , 2009, The European journal of neuroscience.

[14]  M. Duyme,et al.  Sex and performance level effects on brain activation during a verbal fluency task: A functional magnetic resonance imaging study , 2009, Cortex.

[15]  J. McGrath,et al.  Schizophrenia: a concise overview of incidence, prevalence, and mortality. , 2008, Epidemiologic reviews.

[16]  T. Kemper,et al.  The neuropathology of the autism spectrum disorders: what have we learned? , 2008, Novartis Foundation symposium.

[17]  K. Tsutsui.  Neurosteroids in the Purkinje Cell: Biosynthesis, Mode of Action and Functional Significance , 2008, Molecular Neurobiology.

[18]  H. Ackermann.  Cerebellar contributions to speech production and speech perception: psycholinguistic and neurobiological perspectives , 2008, Trends in Neurosciences.

[19]  S. Tobet,et al.  Sex differences in brain developing in the presence or absence of gonads , 2008, Developmental neurobiology.

[20]  P. Witkovsky,et al.  Targeted mutation of the calbindin D28K gene disrupts circadian rhythmicity and entrainment , 2008, The European journal of neuroscience.

[21]  C. Zahn-Waxler,et al.  Disorders of childhood and adolescence: gender and psychopathology. , 2008, Annual review of clinical psychology.

[22]  Yi Zhang,et al.  New Nomenclature for Chromatin-Modifying Enzymes , 2007, Cell.

[23]  S. Tobet,et al.  Neuronal nitric oxide synthase and calbindin delineate sex differences in the developing hypothalamus and preoptic area , 2007, Developmental neurobiology.

[24]  I. Tetko,et al.  Differences in locomotor behavior revealed in mice deficient for the calcium-binding proteins parvalbumin, calbindin D-28k or both , 2007, Behavioural Brain Research.

[25]  Maite Huarte,et al.  The X-Linked Mental Retardation Gene SMCX/JARID1C Defines a Family of Histone H3 Lysine 4 Demethylases , 2007, Cell.

[26]  C. Disteche,et al.  Sex differences in brain expression of X- and Y-linked genes , 2006, Brain Research.

[27]  F. Licastro,et al.  Age‐Related Inflammatory Diseases , 2006 .

[28]  R. Kumar,et al.  Structure, binding interface and hydrophobic transitions of Ca2+-loaded calbindin-D28K , 2006, Nature Structural &Molecular Biology.

[29]  A. Arnold,et al.  Sex Chromosome Complement and Gonadal Sex Influence Aggressive and Parental Behaviors in Mice , 2006, The Journal of Neuroscience.

[30]  L. Garcia-Segura,et al.  Sex differences, developmental changes, response to injury and cAMP regulation of the mRNA levels of steroidogenic acute regulatory protein, cytochrome p450scc, and aromatase in the olivocerebellar system. , 2006, Journal of neurobiology.

[31]  G. Cheron,et al.  Mono‐ and dual‐frequency fast cerebellar oscillation in mice lacking parvalbumin and/or calbindin D‐28k , 2005, The European journal of neuroscience.

[32]  S. Berenbaum,et al.  Prenatal sex hormone effects on child and adult sex-typed behavior: methods and findings , 2005, Neuroscience & Biobehavioral Reviews.

[33]  N. Vesselkin,et al.  Distribution of calcium-binding proteins in the central and peripheral regions of the turtle mesencephalic center torus semicircularis , 2004, Doklady Biological Sciences.

[34]  T. Kosaka,et al.  Expression and possible role of neuronal calcium sensor-1 in the cerebellum , 2004, The Cerebellum.

[35]  M. McCarthy,et al.  Brain estradiol content in newborn rats: sex differences, regional heterogeneity, and possible de novo synthesis by the female telencephalon. , 2004, Endocrinology.

[36]  I. Merchenthaler,et al.  Distribution of estrogen receptor α and β in the mouse central nervous system: In vivo autoradiographic and immunocytochemical analyses , 2004 .

[37]  G. Wooten,et al.  Are men at greater risk for Parkinson’s disease than women? , 2004, Journal of Neurology, Neurosurgery & Psychiatry.

[38]  E. M. Scordalakes,et al.  Aggression and arginine vasopressin immunoreactivity regulation by androgen receptor and estrogen receptor α , 2004, Genes, brain, and behavior.

[39]  D. Pfaff,et al.  Immunolocalization of estrogen receptor β in the mouse brain: Comparison with estrogen receptor α , 2003 .

[40]  A. Konnerth,et al.  Calbindin in Cerebellar Purkinje Cells Is a Critical Determinant of the Precision of Motor Coordination , 2003, The Journal of Neuroscience.

[41]  G. J. de Vries,et al.  Organizational effects of testosterone, estradiol, and dihydrotestosterone on vasopressin mRNA expression in the bed nucleus of the stria terminalis. , 2003, Journal of neurobiology.

[42]  A. Arnold,et al.  Sex differences in mouse cortical thickness are independent of the complement of sex chromosomes , 2003, Neuroscience.

[43]  Robin Lovell-Badge,et al.  A Model System for Study of Sex Chromosome Effects on Sexually Dimorphic Neural and Behavioral Traits , 2002, The Journal of Neuroscience.

[44]  Laura L. Carruth,et al.  Sex chromosome genes directly affect brain sexual differentiation , 2002, Nature Neuroscience.

[45]  P. Thuras,et al.  Purkinje Cell Size Is Reduced in Cerebellum of Patients with Autism , 2002, Cellular and Molecular Neurobiology.

[46]  R. Stornetta,et al.  Vesicular glutamate transporter DNPI/VGLUT2 is expressed by both C1 adrenergic and nonaminergic presympathetic vasomotor neurons of the rat medulla , 2002, The Journal of comparative neurology.

[47]  P. Dutar,et al.  Decrease in calbindin content significantly alters LTP but not NMDA receptor and calcium channel properties , 2002, Neuropharmacology.

[48]  P. Chambon,et al.  Effect of single and compound knockouts of estrogen receptors alpha (ERalpha) and beta (ERbeta) on mouse reproductive phenotypes. , 2000, Development.

[49]  Sickel,et al.  Calbindin‐D28k Immunoreactivity is a Marker for a Subdivision of the Sexually Dimorphic Nucleus of the Preoptic Area of the Rat: Developmental Profile and Gonadal Steroid Modulation , 2000, Journal of neuroendocrinology.

[50]  M. Meyer,et al.  Alterations in Purkinje cell spines of calbindin D‐28 k and parvalbumin knock‐out mice , 2000, The European journal of neuroscience.

[51]  E. Lephart,et al.  Dimorphic expression of medial basal hypothalamic-preoptic area calbindin-D(28K) mRNA during perinatal development and adult distribution of calbindin-D(28K) mRNA in Sprague-Dawley rats. , 1999, Brain research. Molecular brain research.

[52]  S. M. Breedlove,et al.  Sexual Differentiation of the Vertebrate Brain: Principles and Mechanisms , 1998, Frontiers in Neuroendocrinology.

[53]  K. Schilling,et al.  Subcellular localization of specific mRNAs and their protein products in Purkinje cells by combined fluorescence in situ hybridization and immunocytochemistry , 1997, Histochemistry and Cell Biology.

[54]  O. Garaschuk,et al.  Ataxia and altered dendritic calcium signaling in mice carrying a targeted null mutation of the calbindin D28k gene. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[55]  S. Christakos,et al.  Regulation by estrogen through the 5'-flanking region of the mouse calbindin-D28k gene. , 1995, Molecular endocrinology.

[56]  F. Amenta,et al.  Calbindin D-28k immunoreactivity in the rat cerebellar cortex: age-related changes , 1994, Neuroscience Letters.

[57]  G. J. de Vries,et al.  Sex differences in the effects of testosterone and its metabolites on vasopressin messenger RNA levels in the bed nucleus of the stria terminalis of rats , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[58]  M. Somerville,et al.  Reduction of calbindin-28k mRNA levels in Alzheimer as compared to Huntington hippocampus. , 1993, Brain research. Molecular brain research.

[59]  Masahiko Watanabe,et al.  Localization of gene expression of calbindin in the brain of adult rats , 1992, Neuroscience Letters.

[60]  I. Black,et al.  Sexually dimorphic expression of the NGF receptor gene in the developing rat brain , 1991, Brain Research.

[61]  M. Celio,et al.  Calbindin D-28k and parvalbumin in the rat nervous system , 1990, Neuroscience.

[62]  C. Andressen,et al.  Calbindin D‐28k Protein and mRNA Localization in the Rat Brain , 1990, The European journal of neuroscience.

[63]  S. Christakos,et al.  Calcium binding protein (calbindin-D28k) gene expression in the developing and aging mouse cerebellum. , 1990, Brain research. Molecular brain research.

[64]  P. Emson,et al.  Calbindin D28K as a marker for the degeneration of the striatonigral pathway in Huntington's disease , 1990, Brain Research.

[65]  S. Christakos,et al.  Specific reduction of calcium-binding protein (28-kilodalton calbindin-D) gene expression in aging and neurodegenerative diseases. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[66]  L. Cahill,et al.  Sex differences in molecular neuroscience: from fruit flies to humans , 2010, Nature Reviews Neuroscience.

[67]  M. McCarthy,et al.  Steroids, sex and the cerebellar cortex: implications for human disease , 2008, The Cerebellum.

[68]  Enrico Bastianelli,et al.  Distribution of calcium-binding proteins in the cerebellum , 2008, The Cerebellum.

[69]  F. Licastro,et al.  Age-related inflammatory diseases: role of genetics and gender in the pathophysiology of Alzheimer's disease. , 2006, Annals of the New York Academy of Sciences.

[70]  A. Arnold,et al.  Sexually dimorphic expression of the X-linked gene Eif2s3x mRNA but not protein in mouse brain. , 2006, Gene expression patterns : GEP.

[71]  Y. Shimizu,et al.  Cumulative incidence of childhood autism: a total population study of better accuracy and precision. , 2005, Developmental medicine and child neurology.

[72]  I. Merchenthaler,et al.  Distribution of estrogen receptor alpha and beta in the mouse central nervous system: in vivo autoradiographic and immunocytochemical analyses. , 2004, The Journal of comparative neurology.

[73]  D. Pfaff,et al.  Immunolocalization of estrogen receptor beta in the mouse brain: comparison with estrogen receptor alpha. , 2003, Endocrinology.

[74]  M. Mazzocco,et al.  Advances in research on the fragile X syndrome. , 2000, Mental retardation and developmental disabilities research reviews.