Expression of CD38 in Human Neuroblastoma Sh-SY5Y Cells

Abstract

Human CD38 antigen is a 42–45 kDa type II transmembrane glycoprotein with a short N-terminal cytoplasmic domain and a long C-terminal extracellular region. It is widely expressed in different cell types including thymocytes, activated T cells, and terminally differentiated B cells (plasma cells) and it is involved in cellular proliferation and adhesion. CD38 acts as an ectocyclase that converts NAD⁺ to the Ca²⁺-releasing second messenger cyclic ADP-ribose (cADPR). It has been also demonstrated that increased extracellular levels of NAD⁺ and cADPR are involved in inflammatory diseases and in cellular damage, such as ischemia. In the present study, we have characterized the expression of CD38 in human neuroblastoma SH-SY5Y cell line. All-trans-retinoic acid (ATRA) treatment was used to induce cell differentiation. Our results indicate that: a) even if SH-SY5Y cells have a negative phenotype express CD38 at nuclear level, ATRA treatment does not influence this pattern; b) CD38 localizing to the nucleus may co-localize with p80-coilin positive nuclear-coiled bodies; c) purified nuclei, by Western blot determinations using anti-CD38 antibodies, display a band with a molecular mass of −42 kDa; d) SH-SY5Y cells show nuclear ADP-ribosyl cyclase due to CD38 activity; e) the basal level of CD38 mRNA shows a time-dependent increase after treatment with ATRA. These results suggest that the presence of constitutive fully functional CD38 in the SH-SY5Y nucleus has some important implications for intracellular generation of cADP-ribose and subsequent nucleoplasmic calcium release.

Keywords

CD38 Neuroblastoma all-trans-retinoic acid Cajal Body

References

Zocchi

Franco

Guida

Benatti

Bargellesi

Malavasi

Lee

H.C.

and De Flora

. 1993. A single protein immunologically identified as CD38 displays NAD+ glycohydrolase, ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities at the outer surface of human erythrocytes. Biochem. Biophys. Res. Commun. 196:1459.

Partida-Sanchez

Cockayne

D.A.

Monard

Jacobson

E.L.

Oppenheimer

Garvy

Kusser

Goodrich

Howard

Harmsen

. 2001. Cyclic ADP-ribose production by CD38 regulates intracellular calcium release, extracellular calcium influx and Chemotaxis in neutrophils and is required for bacterial clearance in vivo. Nat. Med. 7:1209.

Galione

Lee

H.C.

and Busa

W.B.

. 1991. Ca2+-induced Ca2+ release in sea urchin egg homogenates: Modulation by cyclic ADP-ribose. Science 253:1143.

Schulz

and Krause

. 2004. Inositol 1,4,5-trisphosphate and its coplayers in the concert of Ca²⁺ signalling - new faces in the line up. Curr. Mol. Medical 4:313.

Mehta

Shahid

and Malavasi

. 1996. Human CD38, a cell-surface protein with multiple functions. FASEB J. 10:1408.

Lamkin

T.J.

Chin

Varvayanis

Smith

J.L.

Sramkoski

R.M.

Jacobberger

J.W.

and Yen

. 2006. Retinoic acid-induced CD38 expression in HL-60 myeloblastic leukemia cells regulates cell differentiation or viability depending on expression levels. J. Cell. Biochem. 97:1328.

Kishimoto

Hoshino

Ohori

Kontani

Nishina

Suzawa

Kato

and Katada

. 1998. Molecular mechanism of human CD38 gene expression by retinoic acid. Identification of retinoic acid response element in the first intron. J. Biol. Chem. 273:15429.

Kramer

Steiner

Fodinger

Fiebiger

Rappersberger

Binder

Hofbauer

and Marberger

. 1995. High expression of a CD38-like molecule in normal prostatic epithelium and its differential loss in benign and malignant disease. J. Urol. 154:1636.

Ceni

Pochon

Brun

Muller-Steffner

Andrieux

Grunwald

Schuber

De Waard

Lund

Villaz

and Moutin

M.J.

. 2003. CD38-dependent ADP-ribosyl cyclase activity in developing and adult mouse brain. Biochem. J. 370:175.

10.

Trubiani

Guarnieri

Orciani

Salvolini

and Di Primio

. 2004. Sphingolipid microdomains mediate CD38 internalization: topography of the endocytosis. Int. J. Immunopathol. Pharmacol. 17:293.

11.

Funaro

Reinis

Trubiani

Santi

Di Primio

and Malavasi

. 1998. CD38 functions are regulated through an internalization step. J. Immunol. 160:2238.

12.

Khoo

K.M.

and Chang

C.F.

. 2002. Identification and characterization of nuclear CD38 in the rat spleen. Int. J. Biochem. Cell. Biol. 34:43.

13.

Trubiani

Guarnieri

Eleuterio

Di Giuseppe

Orciani

Angelucci

and Di Primio

. 2007. Insights into nuclear localization and dynamic association of CD38 in Raji and K562 cells. J. Cell. Biochem. In press.

14.

Khoo

K.M.

Han

M.K.

Park

J.B.

Chae

S.W.

Kim

U.H.

Lee

H.C.

Bay

B.H.

and Chang

C.F.

. 2000. Localization of the cyclic ADP-ribose-dependent calcium signaling pathway in hepatocyte nucleus. J. Biol. Chem. 275:24807.

15.

Sleeman

Lyon

C.E.

Platani

Kreivi

J.P.

and Lamond

A.I.

. 1998. Dynamic interactions between splicing snRNPs, coiled bodies and nucleoli revealed using snRNP protein fusions to the green fluorescent protein. Exp. Cell Res. 243:290.

16.

Dundr

Hebert

M.D.

Karpova

T.S.

Stanek

Shpargel

K.B.

Meier

U.T.

Neugebauer

K.M.

Matera

A.G.

and Misteli

. 2004. In vivo kinetics of Cajal body components. JCB 164:831.

17.

Andrade

L.E.C.

Chan

E.K.L.

Raska

Peebles

C.L.

Roos

and Tan

E.M.

. 1991. Human autoantibody to a novel protein of the nuclear coiled body: Immunological characterization and cDNA cloning of p80 coilin. J. Exp. Med. 173:1407.

18.

Mizuguchi

Otsuka

Sato

Ishii

Kon

Yamada

Nishina

Katada

and Ikeda

. 1995. Neuronal localization of CD38 antigen in the human brain. Brain Res. 697:235.

19.

Otsuka

Mizuguchi

Aizawa

Haga

Sato

Inoya

Namba

Machinami

and Ikeda

. 1994. CD38 immunoreactivity in Alzheimer's neurofibrillary tangles (Abstr.). Brain Pathol. 4:558.

20.

Nicolini

Miloso

Zoia

Di Silvestro

Cavaletti

and Tredici

. 1998. Retinoic acid differentiated SH-SY5Y human neuroblastoma cells: an in vitro model to assess drug neurotoxicity. Anticancer Res. 18:2477.

21.

Miloso

Villa

Crimi

Galbiati

Donzelli

Nicolini

and Tredici

. 2004. Retinoic acid-induced neuritogenesis of human neuroblastoma SH-SY5Y cells is ERK independent and PKC dependent. J. Neurosci. Res. 75:241.

22.

Trubiani

De Fazio

Pieri

Mazzanti

and Di Primio

. 2000. Nuclear matrix provides linkage sites for translocated NF-kappa B: morphological evidence. Histochem. Cell Biol. 113:369.

23.

Budde

Sieg

Braunewell

K.H.

Gundelfinger

E.D.

and Pape

H.C.

. 2000. Ca2+-induced Ca2+ release supports the relay mode of activity in thalamocortical cells. Neuron 26:483.

24.

Verderio

Bruzzone

Zocchi

Fedele

Schenk

De Flora

and Matteoli

. 2001. Evidence of a role for cyclic ADP-ribose in calcium signalling and neurotransmitter release in cultured astrocytes. J. Neurochem. 78:646.

25.

Stamenkovic

Staunton

and Seed

. 1989. Molecular cloning of CD38. In Leukocyte Typing, vol IV: White cell differentiation antigen, ed. Oxford University Press, p. 1.

26.

Jackson

D.G.

and Bell

J.I.

. 1990. Isolation of a cDNA encoding the human CD38 (T10) molecule, a cell surface glycoprotein with an unusual discontinuous pattern of expression during lymphocyte differentiation. J. Immunol. 144:2811.

27.

Nata

Takamura

Karasawa

Kumagai

Hashioka

Tohgo

Yonekura

Takasawa

Nakamura

and Okamtoto

. 1997. Human gene encoding CD38 (ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase): Organization, nucleotide sequence and alternative splicing. Gene 186:295.