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Connexin43 in rat pituitary: localization at pituicyte and stellate cell gap junctions and within gonadotrophs

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Abstract

Immunohistochemical methods were employed to investigate the cellular and ultrastructural localization of the gap junction protein connexin43 (Cx43) in rat pituitary. Western blots of pituitary homogenates probed with anti-Cx43 antibodies showed the presence of Cx43 in both anterior and posterior pituitary lobes. By light microscopy (LM), Cx43-immunoreactive (Cx43-IR) puncta were found in all areas of the posterior lobe, but at greater concentrations in peripheral regions of this structure. By electron microscopy (EM), immunogold labelling for Cx43 was seen at gap junctions between thin cytoplasmic processes of pituicytes. No immunoreactivity was detected in the intermediate lobe. The anterior lobe contained puncta similar to but more sparsely scattered than those in the posterior lobe, and by EM analysis these were demonstrated to correspond to labelled gap junctions between stellate cells. In addition, anti-Cx43 antibodies produced intracellular labelling in a small percentage of endocrine cells, which were distributed throughout the anterior lobe and determined by double immunostaining methods to be cells containing luteinizing hormone. By EM, labelling within these cells was associated with predominantly large secretory granules and other loosely organized organelles. The results indicate that gap junctions in the pituitary are composed of Cx43 and that this or a related protein may have a novel intracellular function within gonadotrophs.

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References

  • Bennett MVL, Goodenough DA (1978) Gap junctions, electrotonic coupling, and intercellular communication. Neurosci Res Prog Bull 16:373–485

    Google Scholar 

  • Bennett MVL, Barrio LC, Bargiello TC, Spray DC, Hertzberg EL, Saez JC (1991) Gap junctions: new tools, new answers, new questions. Neuron 6:305–320

    Google Scholar 

  • Beyer EC, Steinberg TH (1991) Evidence that the gap junction protein connexin43 is the ATP induced pore of mouse macrophages. J Biol Chem 277:7971–7974

    Google Scholar 

  • Beyer EC, Paul DL, Goodenough DA (1987) Connexin43: a protein from rat heart homologous to a gap junction protein from liver. J Cell Biol 105:2621–2629

    Google Scholar 

  • Beyer EC, Kistler J, Paul DL, Goodenough DA (1989) Antisera directed against connexin43 peptides react with a 43-kD protein localized to gap junctions in myocardium and other tissues. J Cell Biol 108:595–605

    Google Scholar 

  • Beyer EC, Paul DL, Goodenough DA (1990) Connexin family of gap junction proteins. J Membr Biol 116:187–194

    Google Scholar 

  • Breckenridge LJ, Almers W (1987a) Currents through the fusion pore that forms during exocytosis of a secretory vesicle. Nature 329:814–817

    Google Scholar 

  • Breckenridge LJ, Almers W (1987b) Final steps in exocytosis observed in a cell with giant secretory granules. Proc Natl Acad Sci USA 84:1945–1949

    Google Scholar 

  • Cazalis M, Dayanithi G, Nordmann JJ (1987) Hormone release from isolated nerve endings of the rat neurohypophysis. J Physiol 390:55–70

    Google Scholar 

  • Childs (Moriarty) GV, Ellison DG, Lorenzen JR, Collins TJ, Schwartz NB (1982) Immunocytochemical studies of gonadotropin storage in developing castration cells. Endocrinology 111:1318–1328

    Google Scholar 

  • Chow RH, von Ruden L, Neher E (1992) Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature 356:60–63

    Google Scholar 

  • Crow DS, Beyer EC, Paul DL, Kobe SS, Lau AF (1990) Phosphorylation of connexin43 gap junction protein in uninfected and rous sarcoma virus transformed mammalian fibroblasts. Mol Cell Biol 10:1754–1763

    Google Scholar 

  • Dacheux F (1984) Subcellular localization of gonadotropic hormones in pituitary cells of the castrated pig with the use of pre- and post-embedding immunocytochemical methods. Cell Tissue Res 236:153–160

    Google Scholar 

  • Dermietzel R, Traub V, Hwong TK, Beyer E, Bennett MVL, Spray DC, Willecke K (1989) Differential expression of three gap junction proteins in developing and mature brain tissues. Proc Natl Acad Sci USA 86:10148–10152

    Google Scholar 

  • Forbes MS, Sperelakis N (1985) Intercalated discs of mammalian heart: a review of structure and function. Tissue Cell 17:605–648

    Google Scholar 

  • Hatton GI (1988) Pituicytes, glia and control of terminal secretion. J Exp Biol 139:67–79

    Google Scholar 

  • Hooper ML, Subak-Sharpe JH (1981) Metabolic cooperation between cells. Int Rev Cytol 69:45–104

    Google Scholar 

  • Horvath E, Kovacs K, Ezrin C (1977) Junctional contract between lactotrophs and gonadotrophs in the rat pituitary. IRCS Med Sci 5:511

    Google Scholar 

  • Inoue K, Kurosumi K (1984) Ultrastructural immunocytochemical localization of LH and FSH in the pituitary of the untreated male rat. Cell Tissue Res 235:77–83

    Google Scholar 

  • Kadle R, Zhang JT, Nicholson BJ (1991) Tissue-specific distribution of differentially phosphorylated forms of Cx43. Mol Cell Biol 11:363–369

    Google Scholar 

  • Kelly RB (1985) Pathways of protein secretion in eukaryotes. Science 230:25–32

    Google Scholar 

  • Kurosumi K (1968) Functional classification of cell types of the anterior pituitary gland accomplished by electron microscopy. Arch Histol Jpn 29:329–362

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lah JJ, Hayes DM, Burry RW (1990) A neutral pH silver development method for the visualization of 1-nanometer gold particles in pre-embedding electron microscopic immunocytochemistry. J Histochem Cytochem 38:503–508

    Google Scholar 

  • Laird DW, Revel J-P (1990) Biochemical and immunochemical analysis of the arrangement of connexin43 in rat heart gap junction membranes. J Cell Sci 97:109–117

    Google Scholar 

  • Larsen WJ (1977) Structural diversity of gap junctions. A review. Tissue Cell 3:373–394

    Google Scholar 

  • Larson DM, Haudenschild CC, Beyer EC (1990) Gap junction messenger RNA expression by vascular wall cells. Circ Res 66:1074–1080

    Google Scholar 

  • Lee CJ, Dayanithi G, Nordmann JJ, Lemos JR (1992) Possible role during exocytosis of a Ca2+-activated channel in neurohypophysial granules. Neuron 8:335–342

    Google Scholar 

  • Lemos JR, Ocorr KA, Nordmann JJ (1989) Possible role for ionic channels in neurosecretory granules of the rat neurohypophysis. In: Oxford G, Armstrong C (eds) Secretion and its control. Rockefeller University Press, New York, pp 333–347

    Google Scholar 

  • Lemos JR, Lee OC, Ocorr KA, Dayanithi G, Nordmann JJ (1991) Possible role for neurosecretory granule channel that resembles gap junctions. Ann NY Acad Sci 635:480–482

    Google Scholar 

  • Luke RA, Beyer EC, Hoyt RH, Saffitz JE (1989) Quantitative analysis of intercellular connections by immunohistochemistry of the cardiac gap junction protein connexin43. Circ Res 65:1450–1457

    Google Scholar 

  • Mazet F, Wittenberg BA, Spray DC (1985) Fate of intercellular junctions in isolated adult rat cardiac cells. Circ Res 56:195–204

    Google Scholar 

  • Micevych PE, Abelson L (1991) Distribution of mRNAs coding for liver and heart gap junction proteins in the rat central nervous system. J Comp Neurol 305:96–118

    Google Scholar 

  • Nagy JI, Yamamoto T, Sawchuk MA, Nance DM, Hertzberg EL (1992) Quantitative immunohistochemical and biochemical correlates of connexin43 localization in rat brain. Glia 51:1–9

    Google Scholar 

  • Soji T, Herbert DC (1989) Intercellular communication between rat anterior pituitary cells. Anat Rec 224:523–533

    Google Scholar 

  • Soji T, Yashiro T, Herbert DC (1990) Intercellular communication within the rat anterior pituitary gland. I. Postnatal development and periodic changes after injection of luteinizing hormone-releasing hormone (LHRH) or testosterone. Anat Rec 226:337–341

    Google Scholar 

  • Soji T, Nishizono H, Yashiro T, Herbert DC (1991) Intercellular communication within the rat anterior pituitary gland. III. Postnatal development and periodic changes of cell-to-cell communications in female rats. Anat Rec 231:351–357

    Google Scholar 

  • Spruce AE, Breckenbridge LJ, Lee AK, Almers W (1990) Properties of the fusion pore that forms during exocytosis of a mast cell secretory vesicle. Neuron 4:643–654

    Google Scholar 

  • Staines WA, Meister B, Melander T, Nagy JI, Hokfelt T (1988) Three-color immunofluorescence histochemistry allowing triple labelling within a single section. J Histochem Cytochem 36:145–151

    Google Scholar 

  • Towbin J, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Uchiyama Y, Nakajima M, Watanabe T, Waguri S, Sato N, Yamamoto M, Hashizume Y, Kominami E (1991) Immunocytochemical localization of cathepsin B in rat anterior pituitary endocrine cells, with special reference to its co-localization with renin and prorenin in gonadotrophs. J Histochem Cytochem 39:1199–1205

    Google Scholar 

  • Valnes K, Brandtzaeg P (1985) Retardation of immunofluorescence fading during microscopy. J Histochem Cytochem 33:755–761

    Google Scholar 

  • Vukelic JI, Yamamoto T, Hertzberg EL, Nagy JI (1991) Depletion of connexin43 immunoreactivity in astrocytes after kainic acid-induced lesions in rat brain. Neurosci Lett 130:120–124

    Google Scholar 

  • Walz W (1989) Role of glial cells in the regulation of the brain ion microenvironment. Prog Neurobiol 33:309–333

    Google Scholar 

  • Walz W, Hertz L (1983) Functional interactions between neurons and astrocytes. II. Potassium homeostasis at the cellular level. Prog Neurobiol 20:133–183

    Google Scholar 

  • Watanabe YG (1986) Sex differences in pituitary LH storage and release in LHRH-stimulated pubertal rats. A correlative immunohistochemical and radioimmunoassay study. Cell Tissue Res 243:509–515

    Google Scholar 

  • Watanabe T, Uchiyama Y, Grube D (1991) Topology of chromogranin A and secretogranin II in the rat anterior pituitary: potential marker proteins for distinct secretory pathways in gonadotrophs. Histochemistry 96:285–293

    Google Scholar 

  • Weinrauder H, Zareba-Kowalska A (1984) Glial fibrillary acidic protein and differentiation of neonatal rat pituicytes in vitro. Cell Tissue Res 238:191–195

    Google Scholar 

  • Yamamoto T, Ochalski A, Hertzberg EL, Nagy JI (1990a) LM and EM immunolocalization of the gap junctional protein connexin43 in rat brain. Brain Res 508:313–319

    Google Scholar 

  • Yamamoto T, Ochalski A, Hertzberg EL, Nagy JI (1990b) On the organization of astrocytic gap junctions in rat brain as suggested by LM and EM immunohistochemistry of connexin43 expression. J Comp Neurol 302:853–883

    Google Scholar 

  • Yamamoto T, Hertzberg EL, Nagy JI (1990c) Epitopes of gap junctional proteins localized in neuronal subsurface cisterns. Brain Res 527:135–139

    Google Scholar 

  • Yamamoto T, Hertzberg EL, Nagy JI (1991) Subsurface cisterns in α-motoneurons of the rat and cat: Immunohistochemical detection with antibodies against connexin32. Synapse 8:119–136

    Google Scholar 

  • Yamamoto T, Katsumata N, Tachibana K, Friesen HG, Nagy JI (1992a) Distribution of a novel peptide in the anterior pituitary, gastric pyloric gland and pancreatic islets of rat. J Histochem Cytochem 40:221–229

    Google Scholar 

  • Yamamoto T, Vukelic J, Hertzberg EL, Nagy JI (1992b) Differential anatomical and cellular patterns of connexin43 expression during postnatal development of rat brain. Dev Brain Res 66:165–180

    Google Scholar 

  • Yancey SB, Biswal S, Revel J-P (1992) Spatial and temporal patterns of distribution of the gap junction protein connexin43 during mouse gastrulation and organogenesis. Development 114:203–212

    Google Scholar 

  • Zimmerberg J, Curran M, Cohen FS, Brodwick M (1985) Simultaneous electrical and optical measurements show that membrane fusion precedes secretory granule swelling during exocytosis of beige mouse mast cells. Proc Natl Acad Sci USA 84:1585–1589

    Google Scholar 

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Authors and Affiliations

  1. Biological Laboratory, Kanagawa Dental College, Yokosuka, Japan

    T. Yamamoto & H. Uemura

  2. Department of Physiology, Faculty of Medicine, University of Manitoba, 770 Bannatyne Ave, R3E OW3, Winnipeg, Manitoba, Canada

    M. Z. Hossain, L. J. Murphy & J. I. Nagy

  3. Departments of Neuroscience, Anatomy and Structural Biology, Albert Einstein, College of Medicine, 10461, Bronx, New York, USA

    E. L. Hertzberg

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  1. T. Yamamoto
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  2. M. Z. Hossain
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  3. E. L. Hertzberg
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  4. H. Uemura
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  5. L. J. Murphy
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  6. J. I. Nagy
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Yamamoto, T., Hossain, M.Z., Hertzberg, E.L. et al. Connexin43 in rat pituitary: localization at pituicyte and stellate cell gap junctions and within gonadotrophs. Histochemistry 100, 53–64 (1993). https://doi.org/10.1007/BF00268878

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