Skip to main content
Erschienen in: Experimental Brain Research 2/2006

01.04.2006 | Research Article

The role of connexins in the differentiation of NT2 cells in Sertoli-NT2 cell tissue constructs grown in the rotating wall bioreactor

verfasst von: R. Shamekh, D. F. Cameron, A. E. Willing, S. Saporta

Erschienen in: Experimental Brain Research | Ausgabe 2/2006

Einloggen, um Zugang zu erhalten

Abstract

Neural transplantation is developing as a successful treatment for neurodegenerative diseases such as Parkinson’s disease. The human Ntera-2/D1 (NT2) cell line is an attractive alternative to the use of human fetal neurons as a cell source for transplantation. We have explored combining NT2 cells, as a neuronal source, and Sertoli cells, which may act as a graft facilitator to enhance neuronal survival and differentiation, and ameliorate the host immune response, into a tissue construct for use in cell replacement therapy for neurodegenerative disease. This Sertoli-NT2-aggregated cell (SNAC) tissue construct is formed in the high aspect ratio vessel (HARV) bioreactor. NT2 cells differentiate to dopaminergic NT2N neurons within the SNAC tissue construct without retinoic acid. We report here that the gap junction protein connexin 43 is decreased among differentiated NT2N neurons. Inhibition of connexin 43 with 18β glycyrrhetinic acid and carbenoxolone, a glycyrrhetinic acid derivative, during formation of the SNAC tissue constructs disrupts the differentiation of NT2 cells. Therefore, connexin 43 is important in the differentiation of NT2 cells in the SNAC tissue construct.
Literatur
Zurück zum Zitat Aberts B, Lewis J, Raff M, Roberts K, Watson JD (2003) Molecular biology of the cell. Garland Publishing Inc, New York Aberts B, Lewis J, Raff M, Roberts K, Watson JD (2003) Molecular biology of the cell. Garland Publishing Inc, New York
Zurück zum Zitat Andrews PW (1984) Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. Dev Biol 103:285–293PubMedCrossRef Andrews PW (1984) Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. Dev Biol 103:285–293PubMedCrossRef
Zurück zum Zitat Bani-Yaghoub M, Bechberger JF, Naus CC (1997) Reduction of connexin43 expression and dye-coupling during neuronal differentiation of human NTera2/clone D1 cells. J Neurosci Res 49:19–31PubMedCrossRef Bani-Yaghoub M, Bechberger JF, Naus CC (1997) Reduction of connexin43 expression and dye-coupling during neuronal differentiation of human NTera2/clone D1 cells. J Neurosci Res 49:19–31PubMedCrossRef
Zurück zum Zitat Bani-Yaghoub M, Bechberger JF, Underhill TM, Naus CC (1999) The effects of gap junction blockage on neuronal differentiation of human NTera2/clone D1 cells. Exp Neurol 156:16–32PubMedCrossRef Bani-Yaghoub M, Bechberger JF, Underhill TM, Naus CC (1999) The effects of gap junction blockage on neuronal differentiation of human NTera2/clone D1 cells. Exp Neurol 156:16–32PubMedCrossRef
Zurück zum Zitat Bjorklund A, Stenevi U (1979) Reconstruction of the nigrostriatal dopamine pathway by intracerebral nigral transplants. Brain Res 177:555–560PubMedCrossRef Bjorklund A, Stenevi U (1979) Reconstruction of the nigrostriatal dopamine pathway by intracerebral nigral transplants. Brain Res 177:555–560PubMedCrossRef
Zurück zum Zitat Boucher S, Bennett SA (2003) Differential connexin expression, gap junction intercellular coupling, and hemichannel formation in NT2/D1 human neural progenitors and terminally differentiated hNT neurons. J Neurosci Res 72:393–404PubMedCrossRef Boucher S, Bennett SA (2003) Differential connexin expression, gap junction intercellular coupling, and hemichannel formation in NT2/D1 human neural progenitors and terminally differentiated hNT neurons. J Neurosci Res 72:393–404PubMedCrossRef
Zurück zum Zitat Bravo-Moreno JF, Diaz-Sanchez V, Montoya-Flores JG, Lamoyi E, Saez JC, Perez-Armendariz EM (2001) Expression of connexin43 in mouse Leydig, Sertoli, and germinal cells at different stages of postnatal development. Anat Rec 264:13–24PubMedCrossRef Bravo-Moreno JF, Diaz-Sanchez V, Montoya-Flores JG, Lamoyi E, Saez JC, Perez-Armendariz EM (2001) Expression of connexin43 in mouse Leydig, Sertoli, and germinal cells at different stages of postnatal development. Anat Rec 264:13–24PubMedCrossRef
Zurück zum Zitat Cai J, Cheng A, Luo Y, Lu C, Mattson MP, Rao MS, Furukawa K (2004) Membrane properties of rat embryonic multipotent neural stem cells. J Neurochem 88:212–226PubMedCrossRef Cai J, Cheng A, Luo Y, Lu C, Mattson MP, Rao MS, Furukawa K (2004) Membrane properties of rat embryonic multipotent neural stem cells. J Neurochem 88:212–226PubMedCrossRef
Zurück zum Zitat Cameron DF, Muffly KE (1991) Hormonal regulation of spermatid binding. J Cell Sci 100(Pt 3):623–633PubMed Cameron DF, Muffly KE (1991) Hormonal regulation of spermatid binding. J Cell Sci 100(Pt 3):623–633PubMed
Zurück zum Zitat Cameron DF, Hushen JJ, Colina L, Mallery J, Willing AE, Sanberg PR, Saporta S (2004) Formation and structure of transplantable tissue constructs generated in simulated microgravity from sertoli cells and neuron precursors. Cell Transplant 13:755–763PubMed Cameron DF, Hushen JJ, Colina L, Mallery J, Willing AE, Sanberg PR, Saporta S (2004) Formation and structure of transplantable tissue constructs generated in simulated microgravity from sertoli cells and neuron precursors. Cell Transplant 13:755–763PubMed
Zurück zum Zitat Chen S, Xianwen C, Dehua X, Zhenguo L, Lingfei X, Smith SW, Zhongcheng Z (2003) Behavioral correction of Parkinsonian rats following the transplantation of immortalized fibroblasts genetically modified with TH and GCH genes. Parkinsonism Relat Disord 9(Suppl 2):S91–S97PubMedCrossRef Chen S, Xianwen C, Dehua X, Zhenguo L, Lingfei X, Smith SW, Zhongcheng Z (2003) Behavioral correction of Parkinsonian rats following the transplantation of immortalized fibroblasts genetically modified with TH and GCH genes. Parkinsonism Relat Disord 9(Suppl 2):S91–S97PubMedCrossRef
Zurück zum Zitat Chung SS, Lee WM, Cheng CY (1999) Study on the formation of specialized inter-Sertoli cell junctions in vitro. J Cell Physiol 181:258–272PubMedCrossRef Chung SS, Lee WM, Cheng CY (1999) Study on the formation of specialized inter-Sertoli cell junctions in vitro. J Cell Physiol 181:258–272PubMedCrossRef
Zurück zum Zitat Cudicini C, Lejeune H, Gomez E, Bosmans E, Ballet F, Saez J, Jegou B (1997) Human Leydig cells and Sertoli cells are producers of interleukins-1 and -6. J Clin Endocrinol Metab 82:1426–1433PubMedCrossRef Cudicini C, Lejeune H, Gomez E, Bosmans E, Ballet F, Saez J, Jegou B (1997) Human Leydig cells and Sertoli cells are producers of interleukins-1 and -6. J Clin Endocrinol Metab 82:1426–1433PubMedCrossRef
Zurück zum Zitat Duval N, Gomes D, Calaora V, Calabrese A, Meda P, Bruzzone R (2002) Cell coupling and Cx43 expression in embryonic mouse neural progenitor cells. J Cell Sci 115:3241–3251PubMed Duval N, Gomes D, Calaora V, Calabrese A, Meda P, Bruzzone R (2002) Cell coupling and Cx43 expression in embryonic mouse neural progenitor cells. J Cell Sci 115:3241–3251PubMed
Zurück zum Zitat Elfgang C, Eckert R, Lichtenberg-Frate H, Butterweck A, Traub O, Klein RA, Hulser DF, Willecke K (1995) Specific permeability and selective formation of gap junction channels in connexin-transfected HeLa cells. J Cell Biol 129:805–817PubMedCrossRef Elfgang C, Eckert R, Lichtenberg-Frate H, Butterweck A, Traub O, Klein RA, Hulser DF, Willecke K (1995) Specific permeability and selective formation of gap junction channels in connexin-transfected HeLa cells. J Cell Biol 129:805–817PubMedCrossRef
Zurück zum Zitat Iacovitti L, Stull ND (1997) Expression of tyrosine hydroxylase in newly differentiated neurons from a human cell line (hNT). Neuroreport 8:1471–1474PubMedCrossRef Iacovitti L, Stull ND (1997) Expression of tyrosine hydroxylase in newly differentiated neurons from a human cell line (hNT). Neuroreport 8:1471–1474PubMedCrossRef
Zurück zum Zitat Lelkes PI, Galvan DL, Hayman GT, Goodwin TJ, Chatman DY, Cherian S, Garcia RM, Unsworth BR (1998) Simulated microgravity conditions enhance differentiation of cultured PC12 cells towards the neuroendocrine phenotype. In Vitro Cell Dev Biol Anim 34:316–325PubMedCrossRef Lelkes PI, Galvan DL, Hayman GT, Goodwin TJ, Chatman DY, Cherian S, Garcia RM, Unsworth BR (1998) Simulated microgravity conditions enhance differentiation of cultured PC12 cells towards the neuroendocrine phenotype. In Vitro Cell Dev Biol Anim 34:316–325PubMedCrossRef
Zurück zum Zitat Leung DS, Unsicker K, Reuss B (2002) Expression and developmental regulation of gap junction connexins cx26, cx32, cx43 and cx45 in the rat midbrain-floor. Int J Dev Neurosci 20:63–75PubMedCrossRef Leung DS, Unsicker K, Reuss B (2002) Expression and developmental regulation of gap junction connexins cx26, cx32, cx43 and cx45 in the rat midbrain-floor. Int J Dev Neurosci 20:63–75PubMedCrossRef
Zurück zum Zitat Ling ZD, Potter ED, Lipton JW, Carvey PM (1998) Differentiation of mesencephalic progenitor cells into dopaminergic neurons by cytokines. Exp Neurol 149:411–423PubMedCrossRef Ling ZD, Potter ED, Lipton JW, Carvey PM (1998) Differentiation of mesencephalic progenitor cells into dopaminergic neurons by cytokines. Exp Neurol 149:411–423PubMedCrossRef
Zurück zum Zitat Low HP, Savarese TM, Schwartz WJ (2001) Neural precursor cells form rudimentary tissue-like structures in a rotating-wall vessel bioreactor. In Vitro Cell Dev Biol Anim 37:141–147PubMedCrossRef Low HP, Savarese TM, Schwartz WJ (2001) Neural precursor cells form rudimentary tissue-like structures in a rotating-wall vessel bioreactor. In Vitro Cell Dev Biol Anim 37:141–147PubMedCrossRef
Zurück zum Zitat Nakase T, Naus CC (2004) Gap junctions and neurological disorders of the central nervous system. Biochim Biophys Acta 1662:149–158PubMedCrossRef Nakase T, Naus CC (2004) Gap junctions and neurological disorders of the central nervous system. Biochim Biophys Acta 1662:149–158PubMedCrossRef
Zurück zum Zitat Olanow CW, Goetz CG, Kordower JH, Stoessl AJ, Sossi V, Brin MF, Shannon KM, Nauert GM, Perl DP, Godbold J, Freeman TB (2003) A double-blind controlled trial of bilateral fetal nigral transplantation in Parkinson’s disease. Ann Neurol 54:403–414PubMedCrossRef Olanow CW, Goetz CG, Kordower JH, Stoessl AJ, Sossi V, Brin MF, Shannon KM, Nauert GM, Perl DP, Godbold J, Freeman TB (2003) A double-blind controlled trial of bilateral fetal nigral transplantation in Parkinson’s disease. Ann Neurol 54:403–414PubMedCrossRef
Zurück zum Zitat Othberg AI, Willing AE, Cameron DF, Anton A, Saporta S, Freeman TB, Sanberg PR (1998) Trophic effect of porcine Sertoli cells on rat and human ventral mesencephalic cells and hNT neurons in vitro. Cell Transplant 7:157–164PubMedCrossRef Othberg AI, Willing AE, Cameron DF, Anton A, Saporta S, Freeman TB, Sanberg PR (1998) Trophic effect of porcine Sertoli cells on rat and human ventral mesencephalic cells and hNT neurons in vitro. Cell Transplant 7:157–164PubMedCrossRef
Zurück zum Zitat Pleasure SJ, Page C, Lee VM (1992) Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons. J Neurosci 12:1802–1815PubMed Pleasure SJ, Page C, Lee VM (1992) Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons. J Neurosci 12:1802–1815PubMed
Zurück zum Zitat Risley MS, Tan IP, Roy C, Saez JC (1992) Cell-, age- and stage-dependent distribution of connexin43 gap junctions in testes. J Cell Sci 103( Pt 1):81–96PubMed Risley MS, Tan IP, Roy C, Saez JC (1992) Cell-, age- and stage-dependent distribution of connexin43 gap junctions in testes. J Cell Sci 103( Pt 1):81–96PubMed
Zurück zum Zitat Roscoe WA, Barr KJ, Mhawi AA, Pomerantz DK, Kidder GM (2001) Failure of spermatogenesis in mice lacking connexin43. Biol Reprod 65:829–838PubMedCrossRef Roscoe WA, Barr KJ, Mhawi AA, Pomerantz DK, Kidder GM (2001) Failure of spermatogenesis in mice lacking connexin43. Biol Reprod 65:829–838PubMedCrossRef
Zurück zum Zitat Saporta S, Willing AE, Colina LO, Zigova T, Milliken M, Daadi MM, Sanberg PR (2000) In vitro and in vivo characterization of hNT neuron neurotransmitter phenotypes. Brain Res Bull 53:263–268PubMedCrossRef Saporta S, Willing AE, Colina LO, Zigova T, Milliken M, Daadi MM, Sanberg PR (2000) In vitro and in vivo characterization of hNT neuron neurotransmitter phenotypes. Brain Res Bull 53:263–268PubMedCrossRef
Zurück zum Zitat Saporta S, Willing AE, Shamekh R, Bickford P, Paredes D, Cameron DF (2004) Rapid differentiation of NT2 cells in Sertoli-NT2 cell tissue constructs grown in the rotating wall bioreactor. Brain Res Bull 64:347–356PubMedCrossRef Saporta S, Willing AE, Shamekh R, Bickford P, Paredes D, Cameron DF (2004) Rapid differentiation of NT2 cells in Sertoli-NT2 cell tissue constructs grown in the rotating wall bioreactor. Brain Res Bull 64:347–356PubMedCrossRef
Zurück zum Zitat SiuYi Leung D, Unsicker K, Reuss B (2001) Gap junctions modulate survival-promoting effects of fibroblast growth factor-2 on cultured midbrain dopaminergic neurons. Mol Cell Neurosci 18:44–55PubMedCrossRef SiuYi Leung D, Unsicker K, Reuss B (2001) Gap junctions modulate survival-promoting effects of fibroblast growth factor-2 on cultured midbrain dopaminergic neurons. Mol Cell Neurosci 18:44–55PubMedCrossRef
Zurück zum Zitat Skinner MK (1993) Secretion of growth factors and other regulatory factors. In: Russell LD, Griswold MD (ed) The Sertoli cell. Cache River Press, Clearwater, pp 237–247 Skinner MK (1993) Secretion of growth factors and other regulatory factors. In: Russell LD, Griswold MD (ed) The Sertoli cell. Cache River Press, Clearwater, pp 237–247
Zurück zum Zitat von Coelln R, Unsicker K, Krieglstein K (1995) Screening of interleukins for survival-promoting effects on cultured mesencephalic dopaminergic neurons from embryonic rat brain. Brain Res Dev Brain Res 89:150–154 von Coelln R, Unsicker K, Krieglstein K (1995) Screening of interleukins for survival-promoting effects on cultured mesencephalic dopaminergic neurons from embryonic rat brain. Brain Res Dev Brain Res 89:150–154
Zurück zum Zitat Wang SS, Good TA (2001) Effect of culture in a rotating wall bioreactor on the physiology of differentiated neuron-like PC12 and SH-SY5Y cells. J Cell Biochem 83:574–584PubMedCrossRef Wang SS, Good TA (2001) Effect of culture in a rotating wall bioreactor on the physiology of differentiated neuron-like PC12 and SH-SY5Y cells. J Cell Biochem 83:574–584PubMedCrossRef
Zurück zum Zitat Widenfalk J, Nosrat C, Tomac A, Westphal H, Hoffer B, Olson L (1997) Neurturin and glial cell line-derived neurotrophic factor receptor-beta (GDNFR-beta), novel proteins related to GDNF and GDNFR-alpha with specific cellular patterns of expression suggesting roles in the developing and adult nervous system and in peripheral organs. J Neurosci 17:8506–8519PubMed Widenfalk J, Nosrat C, Tomac A, Westphal H, Hoffer B, Olson L (1997) Neurturin and glial cell line-derived neurotrophic factor receptor-beta (GDNFR-beta), novel proteins related to GDNF and GDNFR-alpha with specific cellular patterns of expression suggesting roles in the developing and adult nervous system and in peripheral organs. J Neurosci 17:8506–8519PubMed
Zurück zum Zitat Willecke K, Eiberger J, Degen J, Eckardt D, Romualdi A, Guldenagel M, Deutsch U, Sohl G (2002) Structural and functional diversity of connexin genes in the mouse and human genome. Biol Chem 383:725–737PubMedCrossRef Willecke K, Eiberger J, Degen J, Eckardt D, Romualdi A, Guldenagel M, Deutsch U, Sohl G (2002) Structural and functional diversity of connexin genes in the mouse and human genome. Biol Chem 383:725–737PubMedCrossRef
Zurück zum Zitat Willing AE, Othberg AI, Saporta S, Anton A, Sinibaldi S, Poulos SG, Cameron DF, Freeman TB, Sanberg PR (1999a) Sertoli cells enhance the survival of co-transplanted dopamine neurons. Brain Res 822:246–250PubMedCrossRef Willing AE, Othberg AI, Saporta S, Anton A, Sinibaldi S, Poulos SG, Cameron DF, Freeman TB, Sanberg PR (1999a) Sertoli cells enhance the survival of co-transplanted dopamine neurons. Brain Res 822:246–250PubMedCrossRef
Zurück zum Zitat Willing AE, Sudberry JJ, Othberg AI, Saporta S, Poulos SG, Cameron DF, Freeman TB, Sanberg PR (1999b) Sertoli cells decrease microglial response and increase engraftment of human hNT neurons in the hemiparkinsonian rat striatum. Brain Res Bull 48:441–444PubMedCrossRef Willing AE, Sudberry JJ, Othberg AI, Saporta S, Poulos SG, Cameron DF, Freeman TB, Sanberg PR (1999b) Sertoli cells decrease microglial response and increase engraftment of human hNT neurons in the hemiparkinsonian rat striatum. Brain Res Bull 48:441–444PubMedCrossRef
Zurück zum Zitat Zigova T, Barroso LF, Willing AE, Saporta S, McGrogan MP, Freeman TB, Sanberg PR (2000) Dopaminergic phenotype of hNT cells in vitro. Brain Res Dev Brain Res 122:87–90PubMedCrossRef Zigova T, Barroso LF, Willing AE, Saporta S, McGrogan MP, Freeman TB, Sanberg PR (2000) Dopaminergic phenotype of hNT cells in vitro. Brain Res Dev Brain Res 122:87–90PubMedCrossRef
Metadaten
Titel
The role of connexins in the differentiation of NT2 cells in Sertoli-NT2 cell tissue constructs grown in the rotating wall bioreactor
verfasst von
R. Shamekh
D. F. Cameron
A. E. Willing
S. Saporta
Publikationsdatum
01.04.2006
Verlag
Springer-Verlag
Erschienen in
Experimental Brain Research / Ausgabe 2/2006
Print ISSN: 0014-4819
Elektronische ISSN: 1432-1106
DOI
https://doi.org/10.1007/s00221-005-0212-x

Weitere Artikel der Ausgabe 2/2006

Experimental Brain Research 2/2006 Zur Ausgabe

Leitlinien kompakt für die Neurologie

Mit medbee Pocketcards sicher entscheiden.

Seit 2022 gehört die medbee GmbH zum Springer Medizin Verlag

Hirnblutung unter DOAK und VKA ähnlich bedrohlich

17.05.2024 Direkte orale Antikoagulanzien Nachrichten

Kommt es zu einer nichttraumatischen Hirnblutung, spielt es keine große Rolle, ob die Betroffenen zuvor direkt wirksame orale Antikoagulanzien oder Marcumar bekommen haben: Die Prognose ist ähnlich schlecht.

Thrombektomie auch bei großen Infarkten von Vorteil

16.05.2024 Ischämischer Schlaganfall Nachrichten

Auch ein sehr ausgedehnter ischämischer Schlaganfall scheint an sich kein Grund zu sein, von einer mechanischen Thrombektomie abzusehen. Dafür spricht die LASTE-Studie, an der Patienten und Patientinnen mit einem ASPECTS von maximal 5 beteiligt waren.

Schwindelursache: Massagepistole lässt Otholiten tanzen

14.05.2024 Benigner Lagerungsschwindel Nachrichten

Wenn jüngere Menschen über ständig rezidivierenden Lagerungsschwindel klagen, könnte eine Massagepistole der Auslöser sein. In JAMA Otolaryngology warnt ein Team vor der Anwendung hochpotenter Geräte im Bereich des Nackens.

Schützt Olivenöl vor dem Tod durch Demenz?

10.05.2024 Morbus Alzheimer Nachrichten

Konsumieren Menschen täglich 7 Gramm Olivenöl, ist ihr Risiko, an einer Demenz zu sterben, um mehr als ein Viertel reduziert – und dies weitgehend unabhängig von ihrer sonstigen Ernährung. Dafür sprechen Auswertungen zweier großer US-Studien.

Update Neurologie

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.