Altered Excitability of Cultured Chromaffin Cells Following Exposure to Multi-Walled Carbon

Daniela Gavello

Education 7th Jannuary 2010 PhD at the Department of Neuroscience (University of Turin) in the laboratory of Prof. Emilio Carbone following a project focused spontaneous activity and calcium channels in neurons and chromaffin cells.

5th October 2009 Fellowship at the Department of Neuroscience (University of Turin) in the laboratory of Prof. Emilio Carbone following the Nanosafe project focused on possible adverse health effects of some nanoparticles. The project is coordinated by NIS (Centre of Excellence on Nanostrucured Interfaces and Surfaces) and financed by Piedmont Region.

1 st March 2009 Fellowship for five months at the the Department of Neuroscience (University of Turin) in the laboratory of Prof. Emilio Carbone, following a project focused on amperometric recordings of secretion in chromaffin cells, using a diamond microelectrodes device.

1st March 2008 Fellowship for one year at Department of Neuroscience (University of Turin, Italy), following the Nanosafe project about: “Nanoparticles: from their impact on the environment and human health to safer production and usage” directed by Prof. Emilio Carbone. The project, coordinated by NIS (Centre of Excellence on Nanostrucured Interfaces and Surfaces) and financed by Piedmont Region, concerns the possible adverse health reactions induced by some nano-particles (carbon nanotubes and silicon nanoparticles) employed by the industrial partners or released in air by incinerators.

Techniques: . Patch clamp: whole cell and perforated-patch configuration, using an EPC-9 patch clamp amplifier (HEKA electronic, Lambrecht, Germany). Use of Pulse programme for on-line and off-line data analysis. . Measurement of chromaffin cells secretion using two different techniques: membrane capacitance and amperometry . MEA technique for recording extracellular action potentials . PCR, RT-PCR . Use of primary cultures of hippocampal neurons and chromaffin cells and clonal cell lines. . Transfection of primary cultures . Little knowledge about siRNA

October 5, 2007 Master ‘s Degree in Medical Biotechnology at University of Turin (Italy) graduate 110 /110 cum laude.

March 2006 -October 2007 Master’s thesis project at Department of Neuroscience (University of

Turin, Italy) about “Up-regulation of 1H T-type channels and low threshold catecholamine secretion by chronic hypoxia in rat chromaffin cells” supervised by Prof. Emilio Carbone. In this experimental work calcium currents and exocytosis on chromaffin rat cells were recorded. The aim was to focus on different degree of expression of LVA and HVA calcium channels in hypoxic and control cells. In order to confirm the major expression of T calcium channels in hypoxic conditions PCR and RT-PCR analysis were also performed.

Techniques: . patch clamp: whole cell and perforated-patch configuration, using an EPC-9 patch clamp amplifier (HEKA electronic, Lambrecht, Germany). Use of Pulse programme. . Primary tissue cultures (chromaffin and hippocampal cells) and clonal cell lines (PC12) . PCR and RT-PCR. . In addition, patch-clamp experiments on neuronal-like differentiated stem cells were carried out during this period in support of other colleagues. The aim was to evaluate the degree of expression of voltage-gated Na+, K+ and Ca2+ in human mesenchimal stem cells differentiated to assume neuronal phenotypes.

September 13, 2005 Bachelor’s Degree in medical Biotechnology at University of Turin (Italy) graduate 108/110.

March 2005-September 2005 Bachelor thesis’s project at Department of Anatomy and Legal Medicine (University of Turin, Italy) about “Effects of Citron-Kinase gene’s deletion on development of hippocampus in the mouse” directed by Prof. Alessandro Vercelli. In order to understand the different morphology of hippocampus in knock- out and control mice neurolucida programme was used. The work was based on brain’s slices analysis after perfusion of the animal.

Techniques: . Neurolucida programme : drawing hippocampus morphology and count cells. . Immunohistochemistry . Microtome . Mouse’s perfusion

October 2004-December 2004 Training at CeRMS institute (San Giovanni Battista Hospital, Turin) working on diagnosis of linphomas in the molecular biology laboratory directed by Prof. Roberto Chiarle.

Techniques: . PCR . Electrophoresis on agarose and poliacrilammide gel

July 3, 2002 Diploma at Liceo scientifico P. Gobetti (Turin, Italy) in sperimental science class: final evaluation 100/100.

Languages ITALIAN (mother language) ENGLISH June 2000 Examination for PET (graduated Pass with merit). June 2001 Examination for First Certificate in English (graduated C).

National Joint-Colloquium Nis-CavNet meeting. Structure and function of congresses Cav1.2 and Cav1.3 L-type channels. Torino 14th-15th September 2012. Oral communication: “Leptin attenuates the depressive action of hypoxia on neuronal firing in the hippocampus: a microelectrode array study”.

Joint-Colloquium Nis-Centro G. Scansetti. NANOSAFE project. Nanoparticles: from their impact on the environment and human health to safer production and usage. Torino. 6-7th October 2011. Oral communication: “Neurotoxic effects of multiwalled carbon nanotubes on adrenal chromaffin cells functioning”.

62nd National congress of the Italian Society of Physiology. Sorrento (Na). 25-27th September 2011. Poster presentation: “Leptin as a new neuroprotective drug from hypoxic injury in the hippocampus: a MEA study” D. Gavello, V. Carabelli, E. Carbone. Dept Neuroscience, NIS Centre of Excellence, Univ. Turin (Turin)

5th meeting of Young Researchers in Physiology. Sestri Levante (Ge). 8- 10th June 2011. Poster and oral presentation: “Leptin as a new neuroprotective drug from hypoxic injury in the hippocampus: a MEA study” D. Gavello, V. Carabelli, E. Carbone. Dept Neuroscience, NIS Centre of Excellence, Univ. Turin (Turin) Winner of the FIRST PRIZE

10th FISV congress, Riva del Garda; 24-27th September 2008. Poster presentation: “Depolarization-evoked release of catecholamines in WT PC12 and in PC12-Trk cells expressing high levels of REST” D. Gavello, V. Comunanza, E. Carbone, J. Meldolesi, V. Carabelli Dept Neuroscience, NIS Centre of Excellence, Univ. Turin (Turin); Vita-Salute San Raffaele Univ. (Milan)

G. Scansetti, Centre-NIS joint Colloquium: “Carbon nanotubes between toxicity and biomedical applications” Torino, June 27th 2008.

International 17th ISCCB. Rouen, Normandy, (France). July 13-17th. Poster congresses presentation: “Leptin dampens mouse chromaffin cell excitability by a PI3K-mediated increase of BK current tone”. Gavello D, Carbone E, Carabelli V, Vandael D.

Neuroscience 2012, 42st annual meeting of the Society for Neuroscience, held October 13 to 17 in New Orleans. Poster: Leptin attenuates the depressive action of hypoxia on spontaneously firing hippocampal neurons: a microelectrode array study Daniela Gavello, Emilio Carbone, Valentina Carabelli

8th FENS forum of European Neuroscience, 14th-18th July 2012, Barcelona (Spain). Poster presentation: “Leptin attenuates the depressive action of hypoxia on neuronal firing in the hippocampus” Daniela Gavello, Emilio Carbone and Valentina Carabelli.

Neuroscience 2011, 41st annual meeting of the Society for Neuroscience, held November 14 to 18 in Washington DC. Poster presentation: SK channels slow down mouse chromaffin cells firing by accumulating Cav1.3 dependent subthreshold currents sensitive to block by intracellular magnesium. Vandael D. H. F., Gavello D., Zuccotti A., Knipper M., Carabelli V., Striessnig J. & Carbone E.

16th International Symposium on Chromaffin Cell biology, Beijing (China) 11-15 July 2011. Altered excitability and secretion following chronic exposures of chromaffin cells to multi-walled carbon nanotubes. Carbone E., Gavello D., Vandael D., Fubini B., Fenoglio I, and Carabelli V.

7th FENS forum of European Neuroscience, 3th-7th July 2010, Amsterdam, Netherlands. Poster presentation: “Multi-walled carbon nanotubes reduce mouse chromaffin cells excitability by enhancing functional noninactivating BK channels”. Daniela Gavello, Roberta Cesa, Federica Premoselli, Bice Fubini, Ivana Fenoglio, Valentina Carabelli, Emilio Carbone.

Neuroscience 2008, 38th annual meeting of the Society for Neuroscience, held Nov. 15 to 19 in Washington, DC. Poster presentation: “Role of Cav1.2 and Cav1.3 channels in triggering exocytosis in mouse chromaffin cells” D. Gavello, S. Mahapatra, PG Montarolo, J. Striessnig* E. Carbone, V. Carabelli Dept Neuroscience, NIS Centre of Excellence, Univ. Turin (Turin); Italy. *Institute of Pharmacy, Pharmacology and toxicology, A-6020 Innsbruck, Austria.

6th Int. Meeting on susbstrate-integrated microelectrodes, 2008 Poster presentation: “Microelectrode array (MEA) analysis of ion channels controlling spontaneous chromaffin cells electrical activity” A. Marcantoni, V. Carabelli, V. Comunanza, D. Gavello, J. Rojo-Ruiz, E. Carbone.

Advanced workshop: T type channels, from discovery to channelopathies: 25 years of research. Kiev, Ukraine, 5-7 June 2008.

“ Cav3.2 coupling to fast “low-threshold” exocytosis in chromaffin cells”. V. Comunanza, D. Gavello, A. Marcantoni, V. Carabelli, E. Carbone. (Proceedings of the meeting)

Collaborations Collaboration with the group of Prof. B. Fubini, Department of Chemistry, University of Turin, from March 2008. The project is focused on in vitro test to assess the possible toxicity of nanoparticles in mouse chromaffin cells.

Collaboration with Prof. J. Meldolesi, Vita-Salute S. Raffaele University, Scientific Institute S. Raffaele, Research Unit of Molecular Neuroscience, Milano, from December 2007 to december 2009. The work was focused on characterization of PC12 cells and rapid neurite outgrowth depending on Rac and enlargosome exocytosis.

Collaboration with the group of Prof. Marlies Knipper, University of Tuebingen (Germany), ENT Clinic; 19-30 of January 2009. Transfection of Cav1.3 channel linked to GFP in mouse chromaffin cells.

Collaboration with the group of Prof. Matteo Mangoni in Montpellier (France); institut de Genomique Fonctionnelle (IGF), CNRS. Genotyping of Cav1.3 knock-out mice: setting up protocols in order to restore the viability of the genetically modified animals.

Collaboration with the group of Prof. J. Striessnig in Innsbruck (Austria): real-time PCR on samples used for siRNA : the aim was to silence the Cav 1.3 gene.

Collaboration with the group of Prof. A. Gasco, Dipartimento di scienza e tecnologia del farmaco, University of Turin, from april 2009. In vitro tests on chromaffin cells cultures to assess the neuroprotective effect of some carnosine derived compounds.

Publications Inhibition of catecholamine secretion by iron-rich and iron-deprived multiwalled carbon nanotubes in chromaffin cells.

Gavello D, Fenoglio I, Fubini B, Cesano F, Premoselli F, Renna A, Carbone E, Carabelli V.

Neurotoxicology. 2013 Aug 30;39C:84-94. doi: 10.1016/j.neuro.2013.08.008. [Epub ahead of print]

Leptin counteracts the hypoxia-induced inhibition of spontaneously firing hippocampal neurons: a microelectrode array study

Daniela Gavello, Jonathan Rojo-Ruiz, Andrea Marcantoni, Claudio Franchino, Emilio Carbone, Valentina Carabelli.

PLoS One 2012. 7(7):e41530.

Altered excitability of cultured chromaffin cells following exposure to multi-walled carbon nanotubes.

Gavello D, Vandael DH, Cesa R, Premoselli F, Marcantoni A, Cesano F, Scarano D, Fubini B, Carbone E, Fenoglio I, Carabelli V.

Nanotoxicology. 2012; 6:47-60

The effect of CdSe-ZnS quantum dots on calcium currents and catecholamine secretion in mouse chromaffin cells.

Gosso S, Gavello D, Giachello CN, Franchino C, Carbone E, Carabelli V. Biomaterials. 2011;32(34):9040-50.

Synthesis, physico-chemical characterization and biological properties of new carnosine amides stable in human serum.

M. Bertinaria, B. Rolando, M. Giorgis, G. Montanaro, M. F. Buonsanti, V. Carabelli, D. Gavello, P.G. Daniele, R. Fruttero, A. Gasco.

J Med Chem 2011.;54(2):611-21.

CaV1.3 as pacemaker channels in adrenal chromaffin cells: specific role on exo- and endocytosis? Comunanza V, Marcantoni A, Vandael DH, Mahapatra S, Gavello D, Carabelli V, Carbone E.

Channels (Austin).2010 4(6):440-6.

Rapid neurite outgrowth in neurosecretory cells and neurons is sustained by the exocytosis of a cytoplasmic organelle, the enlargeosome.

G. Racchetti, A. Lorusso, C. Schulte, D. Gavello, V. Carabelli, R. D’Alessandro and J. Meldolesi.

J. Cell. Sci. 2010. 165-70

Teaching activity I am “cultrice della materia” from 2010. I am a teaching assistant at the Faculty of Pharmacy, for the course of General Physiology and Animal Biology. From 2011 I undertake teaching duties for the course of Animal Biology at the Faculty of Pharmacy.

Courses Management of Innovation. Course promoted by the Unione Industriale of Torino. 22-26th October 2012. Torino, Scuola Assocam Camerana.

Contacts Prof. Emilio Carbone Universitiy of Turin Department of Neuroscience CNISM Research Unit N.I.S. Centre of Excellence Corso Raffaello 30 10125 - Torino (Italy) Phone +39.011.670.7786 Fax +39.011.670.8174 Mobile +39.348.739.8305 e-mail [email protected]

http://www.personalweb.unito.it/emilio.carbone/ http://www.nis.unito.it http://www.cav-net.org

Prof. Valentina Carabelli University of Turin Department of Neuroscience CNISM Research Unit N.I.S. Centre of Excellence Corso Raffaello 30 10125 - Torino (Italy) tel. 0039 011 670 7702 e-mail: [email protected]