Transcriptome-Wide Identification of Preferentially Expressed Genes in the Hypothalamus and Pituitary Gland

To identify preferentially expressed genes in the central endocrine organs of the hypothalamus and pituitary gland, we generated transcriptome-wide mRNA profiles of the hypothalamus, pituitary gland, and parietal cortex in male mice (12–15 weeks old) using serial analysis of gene expression (SAGE). Total counts of SAGE tags for the hypothalamus, pituitary gland, and parietal cortex were 165824, 126688, and 161045 tags, respectively. This represented 59244, 45151, and 55131 distinct tags, respectively. Comparison of these mRNA profiles revealed that 22 mRNA species, including three potential novel transcripts, were preferentially expressed in the hypothalamus. In addition to well-known hypothalamic transcripts, such as hypocretin, several genes involved in hormone function, intracellular transduction, metabolism, protein transport, steroidogenesis, extracellular matrix, and brain disease were identified as preferentially expressed hypothalamic transcripts. In the pituitary gland, 106 mRNA species, including 60 potential novel transcripts, were preferentially expressed. In addition to well-known pituitary genes, such as growth hormone and thyroid stimulating hormone beta, a number of genes classified to function in transport, amino acid metabolism, intracellular transduction, cell adhesion, disulfide bond formation, stress response, transcription, protein synthesis, and turnover, cell differentiation, the cell cycle, and in the cytoskeleton and extracellular matrix were also preferentially expressed. In conclusion, the current study identified not only well-known hypothalamic and pituitary transcripts but also a number of new candidates likely to be involved in endocrine homeostatic systems regulated by the hypothalamus and pituitary gland.

[1]  T. Kumanishi,et al.  cDNA cloning and mRNA expression analysis of the human neuronatin. High level expression in human pituitary gland and pituitary adenomas. , 1997, Journal of molecular neuroscience : MN.

[2]  F. Liu,et al.  Grb10: more than a simple adaptor protein. , 2004, Frontiers in bioscience : a journal and virtual library.

[3]  J. St-Amand,et al.  Characterization of control and immobilized skeletal muscle: an overview from genetic engineering , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[4]  E. Sausville,et al.  dlk, a putative mammalian homeotic gene differentially expressed in small cell lung carcinoma and neuroendocrine tumor cell line. , 1993, The Journal of biological chemistry.

[5]  S. Kawamoto,et al.  Expression profile of active genes in the human pituitary gland. , 2002, Journal of molecular endocrinology.

[6]  D. H. Olster,et al.  Progesterone facilitation of lordosis in male and female Sprague-Dawley rats following priming with estradiol pulses , 1988, Hormones and Behavior.

[7]  T. Branchek,et al.  Localization of messenger RNAs encoding three GABA transporters in rat brain: an in situ hybridization study. , 1995, Brain research. Molecular brain research.

[8]  S. Melmed,et al.  Proteasome inhibitors induce apoptosis in growth hormone- and prolactin-secreting rat pituitary tumor cells. , 2002, The Journal of endocrinology.

[9]  Zhao-Xue Yu,et al.  Lack of Huntingtin-Associated Protein-1 Causes Neuronal Death Resembling Hypothalamic Degeneration in Huntington's Disease , 2003, The Journal of Neuroscience.

[10]  A. Lania,et al.  G protein mutations in endocrine diseases. , 2001, European journal of endocrinology.

[11]  J. Warrington,et al.  Comparison of human adult and fetal expression and identification of 535 housekeeping/maintenance genes. , 2000, Physiological genomics.

[12]  P. Bovolenta,et al.  Six9 (Optx2), a new member of the Six gene family of transcription factors, is expressed at early stages of vertebrate ocular and pituitary development , 1999, Mechanisms of Development.

[13]  D. Dix,et al.  Gene expression patterns associated with infertility in humans and rodent models. , 2004, Mutation research.

[14]  Martin M Matzuk,et al.  Analysis of Ovarian Gene Expression in Follicle-Stimulating Hormone β Knockout Mice. , 2001, Endocrinology.

[15]  D. Gaudet,et al.  The pleiotropic expression of the myotonic dystrophy protein kinase gene illustrates the complex relationships between genetic, biological and clinical covariates of male aging , 2002, The aging male : the official journal of the International Society for the Study of the Aging Male.

[16]  S. Ren,et al.  Gene expression profiling in the human hypothalamus-pituitary-adrenal axis and full-length cDNA cloning. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Y. Nishida,et al.  The top 10 most abundant transcripts are sufficient to characterize the organs functional specificity: evidences from the cortex, hypothalamus and pituitary gland. , 2005, Gene.

[18]  H. Gardner,et al.  Cloning, characterization, and chromosomal localization of Pnck, a Ca2+/calmodulin-dependent protein kinase , 2000 .

[19]  H. Yokota-Hashimoto,et al.  Involvement of Rab27b in the regulated secretion of pituitary hormones. , 2002, Endocrinology.

[20]  L. Jin,et al.  Aberrant DNA methylation of cyclin D2 and p27 genes in rodent pituitary tumor cell lines correlates with specific gene expression , 2000, Endocrine pathology.

[21]  D. Hopkins,et al.  Differential distribution of butyrylcholinesterase and acetylcholinesterase in the human thalamus , 2003, The Journal of comparative neurology.

[22]  E. Van Obberghen,et al.  The Matricellular Protein SPARC/Osteonectin as a Newly Identified Factor Up-regulated in Obesity* , 2001, The Journal of Biological Chemistry.

[23]  R. Siegel,et al.  In vitro conversion of pregnenolone to progesterone by discrete brain areas of the male rat. , 1980, Journal of steroid biochemistry.

[24]  K. Burns,et al.  Analysis of ovarian gene expression in follicle-stimulating hormone beta knockout mice. , 2001, Endocrinology.

[25]  W. Tsang,et al.  Molecular cloning of a novel mRNA (neuronatin) that is highly expressed in neonatal mammalian brain. , 1994, Biochemical and biophysical research communications.

[26]  F. Liu,et al.  Negative regulation of insulin-stimulated mitogen-activated protein kinase signaling by Grb10. , 2004, Molecular endocrinology.

[27]  P. Kelly,et al.  Grb10 Identified as a Potential Regulator of Growth Hormone (GH) Signaling by Cloning of GH Receptor Target Proteins* , 1998, The Journal of Biological Chemistry.

[28]  E. Snyder,et al.  Reproducibility, bioinformatic analysis and power of the SAGE method to evaluate changes in transcriptome , 2005, Nucleic acids research.

[29]  N. Nelson,et al.  The Family of Na+/Cl− Neurotransmitter Transporters , 1998, Journal of neurochemistry.

[30]  Ji Huang,et al.  [Serial analysis of gene expression]. , 2002, Yi chuan = Hereditas.

[31]  G. Kelsey,et al.  DDC and COBL, flanking the imprinted GRB10 gene on 7p12, are biallelically expressed , 2002, Mammalian Genome.

[32]  R. Fleischmann,et al.  Initial assessment of human gene diversity and expression patterns based upon 83 million nucleotides of cDNA sequence. , 1995, Nature.

[33]  S. O’Rahilly,et al.  Identification of the Global Transcriptomic Response of the Hypothalamic Arcuate Nucleus to Fasting and Leptin , 2010, Journal of neuroendocrinology.

[34]  K. Tasanen,et al.  A single polypeptide acts both as the beta subunit of prolyl 4-hydroxylase and as a protein disulfide-isomerase. , 1987, The Journal of biological chemistry.

[35]  Y. Nishida,et al.  Sexually dimorphic gene expression in the hypothalamus, pituitary gland, and cortex. , 2005, Genomics.

[36]  T. Sakurai Orexin: a link between energy homeostasis and adaptive behaviour. , 2003, Current opinion in clinical nutrition and metabolic care.

[37]  S. Nakai,et al.  Calbindin-D28k and calretinin in the rat posterior pituitary; Light and electron microscopic localization and upregulation with dehydration , 2000, Journal of neurocytology.

[38]  Hiroaki Tanaka,et al.  Regulation of Pituitary Gene Expression by Adrenalectomy , 2009, Obesity.

[39]  M. Low,et al.  Leptin activates anorexigenic POMC neurons through a neural network in the arcuate nucleus , 2001, Nature.

[40]  P. J. Larsen,et al.  Hypothalamic CART is a new anorectic peptide regulated by leptin , 1998, Nature.

[41]  Thomas M. Harris,et al.  Use of Laser-Capture Microdissection for the Identification of Marker Genes for the Ventromedial Hypothalamic Nucleus , 2005, The Journal of Neuroscience.

[42]  E. Sage,et al.  SPARC-null mice exhibit increased adiposity without significant differences in overall body weight , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[43]  L. O’Driscoll Gene Expression Profiling , 2011, Methods in Molecular Biology.

[44]  T. Ueno,et al.  Expression patterns of claudin family of tight junction membrane proteins in developing mouse submandibular gland , 2004, Developmental dynamics : an official publication of the American Association of Anatomists.

[45]  L. Alhonen,et al.  Activation of Polyamine Catabolism Profoundly Alters Tissue Polyamine Pools and Affects Hair Growth and Female Fertility in Transgenic Mice Overexpressing Spermidine/SpermineN 1-Acetyltransferase* , 1997, The Journal of Biological Chemistry.

[46]  S. Yen,et al.  Neurosteroidogenesis in astrocytes, oligodendrocytes, and neurons of cerebral cortex of rat brain. , 1999, Endocrinology.

[47]  Y. Nishida,et al.  Regulation of hypothalamic gene expression by glucocorticoid: implications for energy homeostasis. , 2006, Physiological genomics.

[48]  S. Snyder,et al.  A huntingtin-associated protein enriched in brain with implications for pathology , 1995, Nature.

[49]  Mitsuru Nishiyama,et al.  GABA Release from Proopiomelanocortin Neurons , 2004, The Journal of Neuroscience.

[50]  K. Matsumoto,et al.  Purification and Cloning of Hepatocyte Growth Factor Activator Inhibitor Type 2, a Kunitz-type Serine Protease Inhibitor* , 1997, The Journal of Biological Chemistry.

[51]  A. Persechini,et al.  The EF-hand family of calcium-modulated proteins , 1989, Trends in Neurosciences.

[52]  K. Kaibuchi,et al.  Small GTP-binding proteins. , 1992, International review of cytology.

[53]  H. Miyaoka,et al.  Annexin 5 Messenger Ribonucleic Acid Expression in Pituitary Gonadotropes Is Induced by Gonadotropin-Releasing Hormone (GnRH) and Modulates GnRH Stimulation of Gonadotropin Release , 2002, Neuroendocrinology.

[54]  S. Altschul,et al.  SAGEmap: a public gene expression resource. , 2000, Genome research.

[55]  Jacob Jelsing,et al.  Gene expression profiling of individual hypothalamic nuclei from single animals using laser capture microdissection and microarrays , 2009, Journal of Neuroscience Methods.

[56]  I. Pastan,et al.  The nucleotide sequence of a human cellular thyroid hormone binding protein present in endoplasmic reticulum. , 1987, The Journal of biological chemistry.

[57]  S. O’Rahilly,et al.  Novel Leptin-Regulated Genes Revealed by Transcriptional Profiling of the Hypothalamic Paraventricular Nucleus , 2008, The Journal of Neuroscience.

[58]  M. Therrien,et al.  Ptx1, a bicoid-related homeo box transcription factor involved in transcription of the pro-opiomelanocortin gene. , 1996, Genes & development.

[59]  K. Polonsky,et al.  A 48-hour exposure of pancreatic islets to calpain inhibitors impairs mitochondrial fuel metabolism and the exocytosis of insulin. , 2003, Metabolism: clinical and experimental.

[60]  David J. Anderson,et al.  Mammalian achaete-scute homolog 1 is required for the early development of olfactory and autonomic neurons , 1993, Cell.

[61]  K. Lesch,et al.  The brain‐specific protein MLC1 implicated in megalencephalic leukoencephalopathy with subcortical cysts is expressed in glial cells in the murine brain , 2003, Glia.

[62]  F E Bloom,et al.  Overview of the most prevalent hypothalamus-specific mRNAs, as identified by directional tag PCR subtraction. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[63]  Jon T. Willie,et al.  Narcolepsy in orexin Knockout Mice Molecular Genetics of Sleep Regulation , 1999, Cell.

[64]  H. Schulman,et al.  The multifunctional Ca2+/calmodulin-dependent protein kinases. , 1993, Current opinion in cell biology.