NAME:  Daniele Zacchetti

BORN:  Udine, Italy. November 9th, 1964

NATIONALITY: Italian

STATUS:  Married, three children

WORK ADDRESS:
Cellular Neurophysiology Unit
Dibit - San Raffaele Scientific Institute
via Olgettina 58, 20132 Milano - Italy
phone +39-02-2643.4812
fax  +39-02-2643.4813
e-mail: zacchetti.daniele@hsr.it

EDUCATIONS AND QUALIFICATIONS

School Certificate "Maturità Classica", Liceo Classico G. Parini,  Milano, Italy. July 1983. Main subjects: Italian, Latin, ancient Greek language and literature; History and History of Philosophy; elements of Physics, Chemistry, Biology and Mathematics.

Doctorate in Pharmaceutical Chemistry and Technology, grade 110/110 cum laude, University of Milano, Italy. 12 July 1989. Thesis: "Mechanisms of alpha-latrotoxin action: role of Ca2+ in receptor binding and polyphosphoinositide hydrolysis".

Italian State Professional Examination for Pharmacist, Milano, Italy. April 1990.

Ph.D. in Cellular and Molecular Biology, University of Milano, Italy. 14 September 1993. Thesis: "Receptor activation and calcium increases: multiple mechanisms working in PC12 cells".

RESEARCH   EXPERIENCE

Mar 1987 - Jul 1989. Undergraduate student, Cellular Pharmacology Laboratory (Prof. Jacopo Meldolesi), San Raffaele Scient. Inst. and Dept. of Pharmacology, School of Medicine, University of Milano, Milano, Italy.

Aug 1989 - Nov 1991. Research fellow (Prof. Jacopo Meldolesi and Prof. Fabio Grohovaz), CNR Center of Cytopharmacology, Dept. of Pharmacology and San Raffaele Scient. Inst., University of Milano, Milano, Italy.

Nov 1991 - Dec 1994. Junior scientist at the Cellular Neurophysiology Unit (Prof. Fabio Grohovaz), Department of Biological and Technological Science (Dibit) - San Raffaele Scient. Inst., Milano, Italy
(Mar 1992 - Mar 1993: Military Service)

Jan 1995 - Sep 1997. Post-doctoral fellow (Prof. Kai Simons), Cell Biology Programme, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.

Oct 1997 - Present. Staff scientist and project leader at the Department of Neuroscience (Dibit)- San Raffaele Scient. Inst., Milano, Italy.

TOPICS OF INVESTIGATION

Signal transduction of sphingoid molecules. Sphingosylphosphocholine (SPC) and its role in the etiopathogenesis of Niemann-Pick type A Disease.

Translational control of gene expression. Molecular mechanisms of the regulation of translation at the level of cap-dependent translation initiation and internal ribosome entry sites (IRES).

Alzheimer's Disease. Characterization of BACE1 and its role in the etiopathogenesis of Alzheimer's Disease.

Molecular mechanisms of astrocyte activation. Astrocytosis and its role in inflammation and neurodegeneration.

TECHNICAL EXPERTISE

Biochemistry.  Homogeneization of cells and tissues and subcellular fractionation. Separation of proteins by cromatography or mono- and bi-dimensional electroforesis. Production and purification of soluble and membrane proteins: alfa-latrotoxin from L. mactans, recombinant proteins produced in E. coli, in SL2 cells (Drosophila), and in mammalian cells. Reconstitution of proteins in artificial membranes (liposomes and proteoliposomes). Labelling of proteins (iodination of purified proteins, metabolic labelling and purification by immunoprecipitation). Analysis of second messengers (IP3, cAMP, Ca2+...) and secretion of neurotrasmitters.

Molecular Biology. Manipulation and analysis of nucleic acids (RNA e DNA): PCR, RT-PCR, cloning, primer extension, Northern blotting, RNase protection. Gene expression technologies in prokaryots (E. coli) and eukaryots (Drosophila, mammalian cells), including inducibile systems or systems based on viral vectors. Basis of bioinformatics.

Cellular Biology. Cell culture techniques. Computer-assisted optical microscopy (phase contrast, DIC, epifluorescence) in fixed and living cells. Automatic microinjection. Single-cell intracellular Ca2+ analysis by video-imaging.

TEACHING   EXPERIENCE

International School of Biophysics. Nerve-muscle function. Erice, Sicily, Italy. 20 October - 1 November 1991. Lecture: "Calcium fluxes and distribution in neurons".

Ph. D. program in Pharmacology, University of Milano, Italy. Course on "Morphological techniques in pharmacological research". 10 February 1993. Lecture: "Video-imaging".

Demonstrator at the International Course on Advanced techniques for calcium ion measurements in living cells. Milano, Italy. 20-25 September 1993.  Titles: "[Ca2+]i measurements on cell populations" and "Ca2+ fluxes measured by means of 45Ca2+".

Demonstrator at the EMBO Course "Methods in Cell Biology", EMBL, Heidelberg, Germany. 1-11 October 1995. Topic: "Biogenesis of epithelial cell polarity".

Demonstrator at the Ph.D. Program of the European Molecular Biology Laboratory, Heidelberg, Germany. November 1995. Title: "Two-dimensional gel electrophoresis".

School of Physiology and Biophysics. Eterologous expression of membrane proteins. Varese, Italy. 6-9 September 2000. Lecture: "VIP17/MAL, a proteolipid in apical transport vesicles".

Tutor in Molecular and Cellular Biology, Vita-Salute San Raffaele University (a.a. 2000-2001).

Tutor in Cellular Physiology, Vita-Salute San Raffaele University (a.a. 2000-2001).
 
Contract Professor of Pharmacology, Vita-Salute San Raffaele University (a.a. 1999-2000, 2000-2001 e 2001-2002).

Tutor in Biotechnology (Manipulation of nucleic acids), Vita-Salute San Raffaele University (a.a. 2002-2003).

Contract Professor of Biotechnology, Vita-Salute San Raffaele University (a.a. 2002-2003).

OTHER EXPERIENCES

Referee for international peer-reviewed journals (Journal of Cell Biology, Journal of Biological Chemistry, European Journal of Neuroscience)

Referee for  granting agencies (Italian Ministry for Instruction, University and Research)

AWARDS 

Long term EMBO fellowship (1995-1996; declined)

"Marie Curie" Human Capital and Mobility fellowship (1995-1996)

Italian Telethon fellowship (1997)

PUBLICATIONS (Original articles and reviews)

====1999-2003====

De Pietri Tonelli D., Mihailovich M., Schnurbus R., Pesole G., Grohovaz F., and Zacchetti D.
Translational control of Scamper expression via a cell-specific internal ribosome entry site.
Nucl. acids Res. 31: 2508-2513 (2003)

Ehehalt R., Michel B., De Pietri Tonelli D., Zacchetti D., Simons K., and Keller P.
Splice variants of the beta-site APP-cleaving enzyme BACE1 in human brain and pancreas.
Biochem. Biophys. Res. Comm. 293: 30-37 (2002)

Schnurbus R., De Pietri Tonelli D., Grohovaz F., and Zacchetti D.
Re-evaluation of primary structure, topology and localisation of Scamper, a putative intracellular Ca2+ channel activated by sphingosylphosphocholine.
Biochem. J. 362: 183-189 (2002)

Benting J., Lecat S., Zacchetti D., and Simons K.
Protein expression in Drosophila Schneider cells.
Anal. Biochem. 278: 59-68 (2000)

Cheong K.H., Zacchetti D., Schneeberger E.E., and Simons K.
VIP17/MAL, a lipid raft-associated protein, is involved in apical transport in MDCK cells.
Proc.Natl. Acad. Sci.USA 96: 6241-6248 (1999)

====1991-1998====

Bertuzzi F., Zacchetti D., Berra C., Socci C., Pozza G., Pontiroli A.E., and Grohovaz F.
Intercellular Ca2+ waves sustain coordinate insulin secretion in pig islets of Langerhans.
FEBS Lett. 379: 21-25 (1996)

Zacchetti D., Peranen J., Murata M., Fiedler K., and Simons K.
VIP17/MAL, a proteolipid in apical transport vesicles.
FEBS Lett. 377: 465-469 (1995)

Lorenzon P.*, Zacchetti D.*, Codazzi F., Fumagalli G., Meldolesi J., and Grohovaz F.
Ca2+ waves in PC12 neurites: a bidirectional, receptor-oriented form of Ca2+ signalling.
J. Cell Biol. 129:797-804 (1995) [*shared first authorship]

Codazzi F., Menegon A., Zacchetti D., Ciardo A., Grohovaz F., and Meldolesi J.
HIV-1 gp120 glycoprotein induces [Ca2+]i responses not only in type-2 but also in type-1 astrocytes and oligodendrocytes of rat cerebellum.
Europ. J. Neurosci. 7:1333-1341 (1995)

D'Andrea P., Codazzi F., Zacchetti D., Meldolesi J., and Grohovaz F.
Oscillations of cytosolic calcium in rat chromaffin cells: dual modulation in frequency and amplitude.
Biochem. Biophys. Res. Comm. 205: 1264-1269 (1994)

D'Andrea P., Zacchetti D., Meldolesi J., and Grohovaz F.
Mechanism of [Ca2+]i oscillations in rat chromaffin cells: complex Ca2+ dependent regulation of a ryanodine-insensitive oscillator.
J. Biol. Chem. 268: 15213-15220 (1993)

Grohovaz F., Zacchetti D., Lorenzon P., Meldolesi J., and D'Andrea P.
Receptor-mediated intracellular signalling: oscillations and waves of cytosolic calcium.
Biochem. Soc. Trans. 21: 1129-1132 (1993)

Clementi E., Racchetti G., Zacchetti D., Panzeri M.C., and Meldolesi J.
Differential expression of markers and activities in a group of PC12 nerve cell clones.
Europ. J. Neurosci. 4: 944-953 (1992)

Grohovaz F., Zacchetti D., D'Andrea P., Lorenzon P., and Meldolesi J.
[Ca2+]i effects of bradykinin B2 receptor activation in PC12 cells.
Agents and actions 38: 9-15 (1992)

Meldolesi J., Villa A., Podini P., Clementi E., Zacchetti D., D'Andrea P., Lorenzon P., and Grohovaz F.
Intracellular Ca2+ stores in neurons. Identification and functional aspects.
J. Physiol. (Paris) 86: 23-30 (1992).

Clementi E., Sheer H., Zacchetti D., Fasolato C., Pozzan T., and Meldolesi J.
Receptor-activated Ca2+ influx: two independently regulated mechanisms of influx stimulation coexist in neurosecretory PC12 cells.
J. Biol. Chem. 267: 2164-2172 (1992).

Fasolato C., Zottini M., Clementi E., Zacchetti D., Meldolesi J., and Pozzan T.
Intracellular Ca2+ pools in PC12 cells: three intracellular pools are distinguished by their turnover and mechanisms of Ca2+ accumulation, storage, and release.
J. Biol. Chem. 266: 20159-20167 (1991).

Zacchetti D., Clementi E., Fasolato C., Lorenzon P., Zottini M., Grohovaz F., Fumagalli G., Pozzan T., and Meldolesi J.
Intracellular Ca2+ pools in PC12 cells: a unique, rapidly-exchanging pool is sensitive to both inositol 1,4,5-trisphosphate and caffeine-ryanodine.
J. Biol. Chem. 266: 20152-20158 (1991).

Meldolesi J., Clementi E., Fasolato C., Zacchetti D., and Pozzan T.
Ca2+ influx following receptor activation.
Trends in Pharm. Sci. 12: 289-292 (1991).

Grohovaz F., Zacchetti D., Clementi E., Meldolesi J., and Fumagalli G.
Ca2+ imaging in PC12 cells: multiple response patterns to receptor activation reveal new aspects of transmembrane signalling.
J. Cell Biol. 113: 1341-1350 (1991).

Fumagalli G., Zacchetti D., Lorenzon P., and Grohovaz F.
Fluorimetric approaches to the study of calcium transients in living cells.
Cytotechnology 5: 99-102 (1991)

Rosenthal L., Zacchetti D., Madeddu L., and Meldolesi J.
Mode of action of alpha-latrotoxin: role of divalent cations in Ca2+-dependent and Ca2+-independent effects mediated by the toxin.
Mol. Pharmacol. 38: 917-923 (1990).

OTHER PUBLICATIONS

Zacchetti D., and Simons K.
VIP17/MAL and sorting in MDCK cells
Mol. Biol. of the Cell 7(S): 442 (1996)

Fesce R., and Zacchetti D.
Calcium fluxes and distribution in neurons.
In: "Bioelectrochemistry IV. Nerve-muscle function", G. Milazzo, P. Volpe and M. Blank Eds., pp. 53-68, 1994, Plenum Press, New York.

Meldolesi J., Gatti G., Magni M., Clementi E., Zacchetti D., and Scheer H.
Phospholipid hydrolysis and [Ca2+]i control.
In: "Phospholipids and signal transmission", pp. 87-93, 1993, Springer-Verlag, Berlin and Heidelberg.

Clementi E., Sheer H., Zacchetti D., Racchetti G, and Meldolesi J.
PC12 cell clones: experimental tools for studying transmembrane signalling and stores.
Pharmacol. Res. 25(S1): 95-96 (1992).

Zacchetti D., Lorenzon P., Sheer H., Clementi E., Meldolesi J., Fumagalli G., and Grohovaz F.
Receptor activation and Ca2+ homeostasis studied by videoimaging.
Pharmacol. Res. 25(S1): 93-95 (1992)