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Expression of nucleobindin 1 (NUCB1) in pancreatic islets and other endocrine tissues

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Abstract

The protein nucleobindin 1 (NUCB1; also known as CALNUC or Nuc) contains an intriguing combination of DNA- and calcium-binding motifs, a trait that it shares with the protein nucleobindin 2 (NUCB2; also known as nesfatin). NUCB2 has been implicated in several aspects of metabolic control and has been identified in a number of endocrine organs. No such comprehensive mapping of NUCB1 has been presented. We have explored the expression and distribution of NUCB1 in tissues and cells of the mouse endocrine system, with particular focus on the endocrine pancreas. Using reverse transcription plus the polymerase chain reaction (RT-PCR) and Western blot, we demonstrate that NUCB1 is present in the endocrine islets of Langerhans but absent from the exocrine acinar cells. Immunofluorescence studies have revealed that all islet cell types contain NUCB1, including the NUCB2-expressing beta cells. RT-PCR, Western blot and immunofluorescence have shown that NUCB1 is expressed in the pituitary, thyroid, parathyroid, gastrointestinal tract, adrenals and gonads. However, within these tissues, NUCB1 expression is not ubiquitous. For example, in the testis, NUCB1 occurs in the seminiferous tubules but not in the Leydig-cell-containing interstitial tissue. Similarly, the lamina propria of the duodenum lacks NUCB1, despite its presence in enterocytes. Where present, NUCB1 consistently appears to be associated with the Golgi apparatus. Thus, NUCB1 is broadly, but not ubiquitously, expressed in cells of the mouse endocrine system. Together with its location in the Golgi apparatus and its putative Ca2+-binding ability, this distribution suggests a role for NUCB1 in Ca2+ handling/sensing in secretory cells.

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References

  • Abdel-Halim SM, Guenifi A, Luthman H, Grill V, Efendic S, Ostenson CG (1994) Impact of diabetic inheritance on glucose tolerance and insulin secretion in spontaneously diabetic GK-Wistar rats. Diabetes 43:281–288

    Article  PubMed  CAS  Google Scholar 

  • Barnikol-Watanabe S, Gross NA, Gotz H, Henkel T, Karabinos A, Kratzin H, Barnikol HU, Hilschmann N (1994) Human protein NEFA, a novel DNA binding/EF-hand/leucine zipper protein. Molecular cloning and sequence analysis of the cDNA, isolation and characterization of the protein. Biol Chem Hoppe Seyler 375:497–512

    Article  PubMed  CAS  Google Scholar 

  • Bazwinsky-Wutschke I, Wolgast S, Muhlbauer E, Peschke E (2010) Distribution patterns of calcium-binding proteins in pancreatic tissue of non-diabetic as well as type 2 diabetic rats and in rat insulinoma beta-cells (INS-1). Histochem Cell Biol 134:115–127

    Article  PubMed  CAS  Google Scholar 

  • Borgström B, Dahlqvist A, Lundh G, Sjövall J (1957) Studies of intestinal digestion and absorption in the human. J Clin Invest 36:1521–1536

    Article  PubMed  PubMed Central  Google Scholar 

  • Breucker H, Schafer E, Holstein AF (1985) Morphogenesis and fate of the residual body in human spermiogenesis. Cell Tissue Res 240:303–309

    Article  PubMed  CAS  Google Scholar 

  • Chandra S, Kable EP, Morrison GH, Webb WW (1991) Calcium sequestration in the Golgi apparatus of cultured mammalian cells revealed by laser scanning confocal microscopy and ion microscopy. J Cell Sci 100:747–752

    PubMed  Google Scholar 

  • Chen YY, Chan RM, Tan KM, Poh LK, Loke KY, Wang JP, Li H, Hu YH, Wang L, Lee KO, Li GW, Lee YS (2013) The association of a nucleobindin 2 gene (NUCB2) variant with childhood adiposity. Gene 516:48–52

    Article  PubMed  CAS  Google Scholar 

  • Dallas SL, Prideaux M, Bonewald LF (2013) The osteocyte: an endocrine cell … and more. Endocr Rev 34:658–690

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • David V, Hochstenbach F, Rajagopalan S, Brenner MB (1993) Interaction with newly synthesized and retained proteins in the endoplasmic reticulum suggests a chaperone function for human integral membrane protein IP90 (calnexin). J Biol Chem 268:9585–9592

    PubMed  CAS  Google Scholar 

  • de Alba E, Tjandra N (2004) Structural studies on the Ca2+-binding domain of human nucleobindin (calnuc). Biochemistry 43:10039–10049

    Article  PubMed  Google Scholar 

  • Dolensek J, Skelin M, Rupnik MS (2011) Calcium dependencies of regulated exocytosis in different endocrine cells. Physiol Res 60:S29–S38

    PubMed  CAS  Google Scholar 

  • Elayat AA, el-Naggar MM, Tahir M (1995) An immunocytochemical and morphometric study of the rat pancreatic islets. J Anat 186:629–637

    PubMed  PubMed Central  Google Scholar 

  • Foo KS, Brismar H, Broberger C (2008) Distribution and neuropeptide coexistence of nucleobindin-2 mRNA/nesfatin-like immunoreactivity in the rat CNS. Neuroscience 156:563–579

    Article  PubMed  CAS  Google Scholar 

  • Foo KS, Brauner H, Ostenson CG, Broberger C (2010) Nucleobindin-2/nesfatin in the endocrine pancreas: distribution and relationship to glycaemic state. J Endocrinol 204:255–263

    Article  PubMed  CAS  Google Scholar 

  • Garcia-Galiano D, Navarro VM, Roa J, Ruiz-Pino F, Sanchez-Garrido MA, Pineda R, Castellano JM, Romero M, Aguilar E, Gaytan F, Dieguez C, Pinilla L, Tena-Sempere M (2010) The anorexigenic neuropeptide, nesfatin-1, is indispensable for normal puberty onset in the female rat. J Neurosci 30:7783–7792

    Article  PubMed  CAS  Google Scholar 

  • Garcia-Galiano D, Pineda R, Ilhan T, Castellano JM, Ruiz-Pino F, Sanchez-Garrido MA, Vazquez MJ, Sangiao-Alvarellos S, Romero-Ruiz A, Pinilla L, Dieguez C, Gaytan F, Tena-Sempere M (2012) Cellular distribution, regulated expression, and functional role of the anorexigenic peptide, NUCB2/nesfatin-1, in the testis. Endocrinology 153:1959–1971

    Article  PubMed  CAS  Google Scholar 

  • Gonzalez R, Tiwari A, Unniappan S (2009) Pancreatic beta cells colocalize insulin and pronesfatin immunoreactivity in rodents. Biochem Biophys Res Commun 381:643–648

    Article  PubMed  CAS  Google Scholar 

  • Henquin JC, Meissner HP (1984) Significance of ionic fluxes and changes in membrane potential for stimulus-secretion coupling in pancreatic B-cells. Experientia 40:1043–1052

    Article  PubMed  CAS  Google Scholar 

  • Kanuru M, Samuel JJ, Balivada LM, Aradhyam GK (2009) Ion-binding properties of Calnuc, Ca2+ versus Mg2+–Calnuc adopts additional and unusual Ca2+-binding sites upon interaction with G-protein. FEBS J 276:2529–2546

    Article  PubMed  CAS  Google Scholar 

  • Kanuru M, Raman R, Aradhyam GK (2013) Serine protease activity of calnuc: regulation by Zn2+ and G proteins. J Biol Chem 288:1762–1773

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Kapoor N, Gupta R, Menon ST, Folta-Stogniew E, Raleigh DP, Sakmar TP (2010) Nucleobindin 1 is a calcium-regulated guanine nucleotide dissociation inhibitor of G{alpha}i1. J Biol Chem 285:31647–31660

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Kawano J, Kotani T, Ogata Y, Ohtaki S, Takechi S, Nakayama T, Sawaguchi A, Nagaike R, Oinuma T, Suganuma T (2000) CALNUC (nucleobindin) is localized in the Golgi apparatus in insect cells. Eur J Cell Biol 79:208–217

    Article  PubMed  CAS  Google Scholar 

  • Kershaw EE, Flier JS (2004) Adipose tissue as an endocrine organ. J Clin Endocrinol Metab 89:2548–2556

    Article  PubMed  CAS  Google Scholar 

  • Krausz Y, Wollheim CB, Siegel E, Sharp GW (1980) Possible role for calmodulin in insulin release. Studies with trifluoperazine in rat pancreatic islets. J Clin Invest 66:603–607

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Lavoie C, Meerloo T, Lin P, Farquhar MG (2002) Calnuc, an EF-hand Ca(2+)-binding protein, is stored and processed in the Golgi and secreted by the constitutive-like pathway in AtT20 cells. Mol Endocrinol 16:2462–2474

    Article  PubMed  CAS  Google Scholar 

  • Leclerc P, Biarc J, St-Onge M, Gilbert C, Dussault AA, Laflamme C, Pouliot M (2008) Nucleobindin co-localizes and associates with cyclooxygenase (COX)-2 in human neutrophils. PLoS One 3:e2229

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin P, Le-Niculescu H, Hofmeister R, McCaffery JM, Jin M, Hennemann H, McQuistan T, De Vries L, Farquhar MG (1998) The mammalian calcium-binding protein, nucleobindin (CALNUC), is a Golgi resident protein. J Cell Biol 141:1515–1527

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Lin P, Yao Y, Hofmeister R, Tsien RY, Farquhar MG (1999) Overexpression of CALNUC (nucleobindin) increases agonist and thapsigargin releasable Ca2+ storage in the Golgi. J Cell Biol 145:279–289

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Lin P, Fischer T, Weiss T, Farquhar MG (2000) Calnuc, an EF-hand Ca(2+) binding protein, specifically interacts with the C-terminal alpha5-helix of G(alpha)i3. Proc Natl Acad Sci U S A 97:674–679

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Lin P, Li F, Zhang YW, Huang H, Tong G, Farquhar MG, Xu H (2007) Calnuc binds to Alzheimer’s beta-amyloid precursor protein and affects its biogenesis. J Neurochem 100:1505–1514

    PubMed  CAS  Google Scholar 

  • Lin P, Fischer T, Lavoie C, Huang H, Farquhar MG (2009) Calnuc plays a role in dynamic distribution of Galphai but not Gbeta subunits and modulates ACTH secretion in AtT-20 neuroendocrine secretory cells. Mol Neurodegener 4:15

    Article  PubMed  PubMed Central  Google Scholar 

  • Linstedt AD, Hauri HP (1993) Giantin, a novel conserved Golgi membrane protein containing a cytoplasmic domain of at least 350 kDa. Mol Biol Cell 4:679–693

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Mears D (2004) Regulation of insulin secretion in islets of Langerhans by Ca(2+)channels. J Membr Biol 200:57–66

    Article  PubMed  CAS  Google Scholar 

  • Miura K, Titani K, Kurosawa Y, Kanai Y (1992) Molecular cloning of nucleobindin, a novel DNA-binding protein that contains both a signal peptide and a leucine zipper structure. Biochem Biophys Res Commun 187:375–380

    Article  PubMed  CAS  Google Scholar 

  • Miura K, Kurosawa Y, Kanai Y (1994) Calcium-binding activity of nucleobindin mediated by an EF hand moiety. Biochem Biophys Res Commun 199:1388–1393

    Article  PubMed  CAS  Google Scholar 

  • Miura K, Hirai M, Kanai Y, Kurosawa Y (1996) Organization of the human gene for nucleobindin (NUC) and its chromosomal assignment to 19q13.2-q13.4. Genomics 34:181–186

    Article  PubMed  CAS  Google Scholar 

  • Mochizuki N, Hibi M, Kanai Y, Insel PA (1995) Interaction of the protein nucleobindin with G alpha i2, as revealed by the yeast two-hybrid system. FEBS Lett 373:155–158

    Article  PubMed  CAS  Google Scholar 

  • Nakata M, Manaka K, Yamamoto S, Mori M, Yada T (2011) Nesfatin-1 enhances glucose-induced insulin secretion by promoting Ca(2+) influx through L-type channels in mouse islet beta-cells. Endocr J 58:305–313

    Article  PubMed  CAS  Google Scholar 

  • Oh IS, Shimizu H, Satoh T, Okada S, Adachi S, Inoue K, Eguchi H, Yamamoto M, Imaki T, Hashimoto K, Tsuchiya T, Monden T, Horiguchi K, Yamada M, Mori M (2006) Identification of nesfatin-1 as a satiety molecule in the hypothalamus. Nature 443:709–712

    Article  Google Scholar 

  • Petersson U, Somogyi E, Reinholt FP, Karlsson T, Sugars RV, Wendel M (2004) Nucleobindin is produced by bone cells and secreted into the osteoid, with a potential role as a modulator of matrix maturation. Bone 34:949–960

    Article  PubMed  CAS  Google Scholar 

  • Pezzati R, Bossi M, Podini P, Meldolesi J, Grohovaz F (1997) High-resolution calcium mapping of the endoplasmic reticulum-Golgi-exocytic membrane system. Electron energy loss imaging analysis of quick frozen-freeze dried PC12 cells. Mol Biol Cell 8:1501–1512

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Pinton P, Pozzan T, Rizzuto R (1998) The Golgi apparatus is an inositol 1,4,5-trisphosphate-sensitive Ca2+ store, with functional properties distinct from those of the endoplasmic reticulum. EMBO J 17:5298–5308

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Ramage L, Junne T, Hahne K, Lithgow T, Schatz G (1993) Functional cooperation of mitochondrial protein import receptors in yeast. EMBO J 12:4115–4123

    PubMed  CAS  PubMed Central  Google Scholar 

  • Riva M, Nitert MD, Voss U, Sathanoori R, Lindqvist A, Ling C, Wierup N (2011) Nesfatin-1 stimulates glucagon and insulin secretion and beta cell NUCB2 is reduced in human type 2 diabetic subjects. Cell Tissue Res 346:393–405

    Article  PubMed  CAS  Google Scholar 

  • Satin LS (2000) Localized calcium influx in pancreatic beta-cells: its significance for Ca2+-dependent insulin secretion from the islets of Langerhans. Endocrine 13:251–262

    Article  PubMed  CAS  Google Scholar 

  • Sooy K, Schermerhorn T, Noda M, Surana M, Rhoten WB, Meyer M, Fleischer N, Sharp GW, Christakos S (1999) Calbindin-D(28 k) controls [Ca(2+)](i) and insulin release. Evidence obtained from calbindin-d(28 k) knockout mice and beta cell lines. J Biol Chem 274:34343–34349

    Article  PubMed  CAS  Google Scholar 

  • Stengel A, Goebel M, Yakubov I, Wang L, Witcher D, Coskun T, Tache Y, Sachs G, Lambrecht NW (2009) Identification and characterization of nesfatin-1 immunoreactivity in endocrine cell types of the rat gastric oxyntic mucosa. Endocrinology 150:232–238

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Stengel A, Goebel M, Tache Y (2011) Nesfatin-1: a novel inhibitory regulator of food intake and body weight. Obes Rev 12:261–271

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Sudhof TC (2012) Calcium control of neurotransmitter release. Cold Spring Harb Perspect Biol 4:a011353

    Article  PubMed  PubMed Central  Google Scholar 

  • Tagaya Y, Osaki A, Miura A, Okada S, Ohshima K, Hashimoto K, Yamada M, Satoh T, Shimizu H, Mori M (2012) Secreted nucleobindin-2 inhibits 3T3-L1 adipocyte differentiation. Protein Pept Lett 19:997–1004

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Tomita T, Doull V, Kimmel JR, Pollock HG (1984) Pancreatic polypeptide and other hormones in pancreas of obese (ob/ob) mice. Diabetologia 27:454–459

    Article  PubMed  CAS  Google Scholar 

  • Vas S, Adori C, Konczol K, Katai Z, Pap D, Papp RS, Bagdy G, Palkovits M, Toth ZE (2013) Nesfatin-1/NUCB2 as a potential new element of sleep regulation in rats. PLoS One 8:e59809

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Wagner L, Oliyarnyk O, Gartner W, Nowotny P, Groeger M, Kaserer K, Waldhausl W, Pasternack MS (2000) Cloning and expression of secretagogin, a novel neuroendocrine- and pancreatic islet of Langerhans-specific Ca2+-binding protein. J Biol Chem 275:24740–24751

    Article  PubMed  CAS  Google Scholar 

  • Wang SN, Miyauchi M, Koshikawa N, Maruyama K, Kubota T, Miura K, Kurosawa Y, Awaya A, Kanai Y (1994) Antigen expression associated with lymph node metastasis in gastric adenocarcinomas. Pathol Int 44:844–849

    Article  PubMed  CAS  Google Scholar 

  • Wendel M, Sommarin Y, Bergman T, Heinegard D (1995) Isolation, characterization, and primary structure of a calcium-binding 63-kDa bone protein. J Biol Chem 270:6125–6133

    Article  PubMed  CAS  Google Scholar 

  • Yanez M, Gil-Longo J, Campos-Toimil M (2012) Calcium binding proteins. Adv Exp Med Biol 740:461–482

    Article  PubMed  CAS  Google Scholar 

  • Yang SN, Berggren PO (2006) The role of voltage-gated calcium channels in pancreatic beta-cell physiology and pathophysiology. Endocr Rev 27:621–676

    Article  PubMed  CAS  Google Scholar 

  • Yosten GL, Redlinger L, Samson WK (2012) Evidence for a role of endogenous nesfatin-1 in the control of water drinking. J Neuroendocrinol 24:1078–1084

    Article  PubMed  CAS  Google Scholar 

  • Zhang AQ, Li XL, Jiang CY, Lin L, Shi RH, Chen JD, Oomura Y (2010) Expression of nesfatin-1/NUCB2 in rodent digestive system. World J Gastroenterol 16:1735–1741

    Article  PubMed  CAS  PubMed Central  Google Scholar 

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Acknowledgments

The authors are grateful to Dr. M. Wendel, Dr. S. Efendic and Dr. J.R. Kimmel for the donation of antisera used in this study. They also thank Dr. Tomas Hökfelt (for providing access to equipment) and the Knut and Alice Wallenberg Foundation (CLICK facility). They greatly appreciate the expert technical assistance of Dr. Andrea Dicker with islet isolation and thank Arash Hellysaz and Dr. Teresa Femenia Canto for their help with immunofluorescence and Western blot experiments, respectively.

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Authors and Affiliations

  1. Department of Neuroscience, Karolinska Institutet, 171 77, Stockholm, Sweden

    Paul Williams, Susanne Tulke & Christian Broberger

  2. The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, 171 77, Stockholm, Sweden

    Erwin Ilegems & Per-Olof Berggren

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  1. Paul Williams
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  2. Susanne Tulke
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  3. Erwin Ilegems
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  4. Per-Olof Berggren
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  5. Christian Broberger
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Correspondence to Paul Williams or Christian Broberger.

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This study was generously funded by a Starting Investigator Grant from the European Research Council (ENDOSWITCH) and supported by the Swedish Research Council, the Strategic Research Program in Diabetes at the Karolinska Institutet, the Elin Danielsson’s Foundation and the Novo Nordisk Foundation to C.B. and by grants from the Swedish Diabetes Association, the Family Erling-Persson Foundation, the Skandia Insurance Company, the Novo Nordisk Foundation, the Berth von Kantzow’s Foundation, the Knut and Alice Wallenberg Foundation, the Diabetes and Wellness Foundation and the Stichting af Jochnick Foundation to P.-O.B. P.W. is supported by graduate training faculty funding from the Karolinska Institutet.

P.-O.B. is the founder and chief executive officer of Biocrine, a biotechnology company, and also serves on the company’s Board of Directors. E.I. serves as a consultant for Biocrine.

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Williams, P., Tulke, S., Ilegems, E. et al. Expression of nucleobindin 1 (NUCB1) in pancreatic islets and other endocrine tissues. Cell Tissue Res 358, 331–342 (2014). https://doi.org/10.1007/s00441-014-1948-z

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  • DOI: https://doi.org/10.1007/s00441-014-1948-z

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