Prof. Paola Paggi Università degli Studi di Roma

Prof. Paola Paggi
Università degli Studi di Roma "La Sapienza", Dipartimento di Biologia Cellulare e dello Sviluppo
P.le Aldo Moro 5, 00185 Roma, Italy.
tel.: +39 06 4991 2323; fax: +39 06 4991 2351; e-mail: [email protected]
Full professor of General Physiology, Dipartimento di Biologia Cellulare e dello Sviluppo,
Università "La Sapienza"
Research area: Meccanismi di riconoscimento delle macromolecole di interesse biologico
NEURONAL RESPONSE TO EXPERIMENTAL INTERRUPTION OF THE NEURAL
CIRCUITS: A MOLECULAR AND STRUCTURAL STUDY IN AUTONOMIC GANGLIA
IN VIVO
SUMMARY
The organisation of synaptic contacts in neural circuits bears a basic relevance to the
accomplishment of brain functions, behavioural control and cognitive activities. The formation,
maintenance and remodelling of synaptic contacts are complex processes that occur during
development as well as in mature neurones and are regulated by a cohort of intra and extracellular
events. They reflect the response of neurones to their own bioelectrical activity, to the presence of
extracellular soluble factors and components of the extracellular matrix, to interactions with
neighbouring neurones, glial cells and target organs, and to pathological processes and injuries.
Autonomic ganglia, in the peripheral nervous system, constitute a model system particularly suited
to experimentally approach these complex interactions. Ganglia display rather complex connectivity
and share many features with the CNS. They lack the wide variety of neurotransmitters, neuronal
cell types and circuital variations of the CNS, but on the other hand they have synapses relatively
accessible and the anatomy and pharmacological basis of ganglionic transmission are essentially
well known. Ganglionic neurones can be easily subjected to various patterns of activation, easily
deprived of their neuronal input (denervated), or otherwise injured (axotomy) by simply cutting or
crushing the pre- or post-ganglionic nerve trunks, respectively; reinnervation, regeneration of the
axonal output and recovery of synaptic function can be monitored both in vivo and in vitro. In order
to identify factors and mechanisms controlled by input and output connections in autonomic
ganglia, structural, immunocytochemical and molecular approaches will be combined. Two
different, but complementary, experimental animal models will be used on account of their peculiar
structural and functional organisation: the rodent sympathetic superior cervical ganglion (SCG), and
the quail parasympathetic ciliary ganglion (CG). The project is articulated in two main lines: 1) We
will investigate the molecular mechanisms involved in intraganglionic synapse remodelling,
induced by injury of the SCG and CG nerves, paying particular attention to the role of extracellular
proteases and extracellular matrix proteins; 2) The relevant pathological changes produced by lack
of dystrophin, a protein of the cortical cytoskeleton, will be investigated in mdx mice, an animal
model for Duchenne muscular dystrophy. The following aspects will be analysed: a) large-scale
analysis of gene expression in mdx mouse SCG to search for genes affected by the spontaneous
mutation in the dystrophin gene and possibly responsible for the altered reaction to axotomy of
SCG neurones previously observed; b) analysis of the nicotinic currents in slices of mdx mouse
SCG, in which the nicotinic acetylcholine receptors containing the α3 subunit (α3nAChRs) are
drastically reduced compared with the wild-type; c) damage to SCG muscular target organs and its
relation to the loss of ganglionic neurones observed in adult mdx mice, by measuring the levels of
trophic factors and their receptors; d) the neurone-target interplay, by co-colturing explants of wildtype and mdx mouse SCG with the respective target organs (iris, heart, submandibular gland) or
with those derived from the other strain (wild-type SCG versus mdx target organs and viceversa).
53. Schreiber R.C. Hyatt-Sachs H., Bennett T.A. et al. (1994) Galanin expression increases in adult rat
sympathetic neurons after axotomy. Neuroscience 60:17-27.
54. Song G., Cechvala C., Resnick D.K., et al (2001) Genechip analisys after acute spinal cord injury in rat.
J. Neurochem. 79:804-815.
55. Squitti R., De Stefano M.E., Edgar D. et al. (1999) Effects of axotomy on the expression and
ultrastructural localization of N-cadherin and N-CAM in the quail ciliary ganglion: an in vivo model of
neuroplasticity. Neuroscience 91:707-722.
56. Sugita S., Fumiaki S., Tang J., et al. (2001) A stoichiometric complex of neurexins and dystroglycan in
brain. J. Cell Biol. 154:435-445.
57. Szklarczyk A, Lapinska J, Rylski M, et al. (2002) Matrix metalloproteinase-9 undergoes expression and
activation during dendritic remodeling in adult hippocampus. J. Neurosci. 22:920-930.
58. Toth M, Hernandez-Barrantes S, Osenkowski P, et al. (2002) Complex pattern of membrane type 1
matrix metalloproteinase shedding. Regulation by autocatalytic cells surface inactivation of active
enzyme. J Biol. Chem. 277:26340-50.
59. Werb Z. (1997) ECM and cell surface proteolysis: regulating cellular ecology. Cell 91:439-442.
60. Yamada H., Saito F., Fukuta-Ohi H., et al. (2001) Processing of β-dystroglycan by matrix
metalloproteinase disrupts the link between the etracellular matrix and cell membrane via the
dystroglycan complex. Hum. Mol. Gen. 10:1563-1569.
61. Zaccaria M.L., De Stefano M.E., Gotti C., et al.(2000) Selective reduction in the nicotinic acetylcholine
receptor and dystroglycan at the postsynaptic apparatus of mdx mouse superior cervical ganglion. J.
Neuropathol. Exp. Neurol. 59:103-112.
62. Zaccaria M.L., De Stefano M.E., Properzi F., et al. (1998) Disassembly of the cholinergic postsynaptic
apparatus induced by axotomy in mouse sympathetic neurons: loss of dystrophin and alphadystroglycan immunoreactivity precedes that of the acetylcholine receptor.
63. Zhou Y., Deneris E. and Zigmond R.E. (2001) Nicotinic acetylcholine receptor subunit proteins α7 and
B4 decrease in the superior cervical ganglion after axotomy. J. Neurobiol. 46: 178-192.
Scientific credentials of the participants to the research project
1) Dr. Maria Egle De Stefano, PhD. Researcher in the Department of Cellular and Developmental
Biology, University of Rome "La Sapienza". Previously recipient of a fellowship by Istituto
Pasteur- Fondazione Cenci Bolognetti. Research associate at the University of Connecticut (19911994), Laboratory of Neuromorphology, directed by Dr. Enrico Mugnaini.
2) Dr.Arianna Del Signore, Post-doctoral Fellow in the Department of Cellular and Developmental
Biology, University of Rome "La Sapienza".
3) Dr. Lucia Leone, Post-doctoral Fellow in the Department of Cellular and Developmental
Biology, University of Rome "La Sapienza".
4) Dr. Loredana Lombardi, PhD student in the Department of Cellular and Developmental Biology,
University of Rome "La Sapienza".
National and international collaborations
1) Dr. F. Schroedl, Institute of Anatomy, University of Erlangen-Nurnberg, Erlangen, Germany.
2) Dr. Cecilia Gotti, CNR, Cellular and Molecular Pharmacology Center, Department of
Pharmacology, University of Milan, Italy.
3) Dr. Tamara C. Petrucci, Laboratory of Cell Biology, Istituto Superiore di Sanità, Rome, Italy.
4) Dr. Carla Perrone Capano, School of Pharmacy, Università di Catanzaro Magna Grecia,
Roccelletta Borgia (CZ), Italy.
5) Dr. Ernesto Di Mauro, Department of Genetics and Molecular Biology, University of Rome “La
Sapienza”, Italy.
6) Dr.Alberto Oliverio, Department of Genetics and Molecular Biology, University of Rome “La
Sapienza”, Italy.
7) Dr. Rodolfo Negri, Department of Cellular and Developmental Biology, University of Rome "La
Sapienza", Italy.
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8) Dr Fabrizio Eusebi, Department of Human Physiology and Pharmacology, University of Rome
"La Sapienza", Italy.
9) Dr. Oscar Sacchi, Department of Biology, University of Ferrara, Italy
10) Dr. A. Maria Di Giulio and Dr. Alfredo Gorio, Department of Medicine and Surgery, School of
Medicine, S. Paolo Hospital, University of Milan, Italy.
11) Dr. Riccardo Fesce, Department of Structural and Functional Biology, University of Insubria,
Varese-Como, Italy.
Scientific credentials of Paola Paggi proponent of the research project
Full professor of General Physiology and in charge of the Neurobiology course in the Department
of Cellular and Developmental Biology, University of Rome "La Sapienza". National Coordinator
of MURST Cofin Research Projects. Principal investigator of research programs of Ateneo
(University "La Sapienza"). Visiting Professor Dept. of Genetics and Anatomy, and
Bioarchitectonics Center, School of Medicine, Case Western Reserve University, Cleveland, Ohio
(1982-1992).
List of the last five years publications related to the project
1. De Stefano, M.E., Leone, L. and Paggi, P. (2001) “Polysialylated neural cell adhesion molecule
is involved in the neuroplasticity induced by axonal injury in the avian ciliary ganglion”
Neuroscience 103: 1093-1104.
2. Zaccaria, M.L., Perrone-Capano, C., Melucci-Vigo, G., Gaeta, L., Petrucci, T.C. and Paggi, P.
(2001) “Differential regulation of transcripts for dystrophin isoforms, dystroglycan and α3AChR
subunit in mouse sympathetic ganglia following postganglionic nerve crush”. Neurobiology of
Disease 8: 513-524.
3. Zaccaria, M.L., Di Tommaso, F., Brancaccio, A., Paggi, P. and Petrucci, T.C. (2001)
“Dystroglycan distribution in adult mouse brain: a light and electron microscopy study”
Neuroscience 104: 311-324.
4. Del Signore A., Gotti C., De Stefano M.E., Moretti M. and Paggi P. (2002)"Dystrophin stabilizes
α3- but not α7-containing nicotinic acetylcholine receptor subtypes at the postsynaptic apparatus in
the mouse superior cervical ganglion" Neurobiology of Disease, 10: 54-66.
5. Coccurello R., Castellano C., Paggi P., Mele A., and Oliverio A. (2002) “'Genetically dystrophic
mdx/mdx mice exhibit decreased response to nicotine in passive avoidance” Neuroreport, 13: 12191222.
6. Mandillo S., Del Signore A., Paggi P., Francia N., Santucci D., Mele A., Oliverio A. (2003)
Effects of acute and repeated daily exposure to hypergravity on spatial learning in mice. Neurosci.
Lett. 336: 147-150.
7. Del Signore, A., Gotti, C., Rizzo, A., Moretti, M. and Paggi, P. (2004) Nicotinic acetylcholine
receptor subtypes in the rat sympathetic ganglion: pharmacological characterization, subcellular
distribution and effect of pre-and post-ganglionic nerve crush. J. Neuropathol. Exp. Neurol. 63:
138-150.
8. Del Signore, A., Mandillo, S., Rizzo, A., Di Mauro, E., Mele, A., Negri, R., Oliverio, A. and
Paggi, P. (2004) “Hippocampal gene expression is modulated by hypergravity” Eur. J. Neurosci.19:
667-677.
9. De Stefano, M. E., Leone, L., Lombardi, L. and Paggi, P. (2005)“ Lack of dystrophin leads to the
selective loss of superior cervical ganglion neurons projecting to muscular targets in genetically
dystrophic mdx mice” Neurobiol. Dis. (In Press).
10. Leone, L., De Stefano, M.E., Petrucci, T.C. and Paggi, P. (2005) Axotomy of sympathetic
neurons activates the metalloproteinase-2 pathway. J. Neuropathol. Exp. Neurol. (In Press).
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