1-GENOMICS AND PROTEOMICS OF
REPRODUCTIVE FUNCTIONS
2- IN
VIVO, EX VIVO AND IN VITRO ANIMAL MODELS FOR
INFLAMMATION STUDIES IN SWINE SPECIES
3-
TRANSGENIC PIG PRODUCTION
1- GENOMICS AND PROTEOMICS OF REPRODUCTIVE FUNCTIONS
Experimental approaches:
Female reproduction study; male reproduction study; molecular and
endocrinological analyses for reproductive and metabolic state
assessment; ethological and welfare assessment in pigs; evaluation
of swine embryos gene expression; genomics and proteomics
approaches specific for swine species.
Principal fields of research:
In the ovary, angiogenesis and angioregression
alternatively appears. In this physiological model we study
the phenomenon at gene expression and regulation levels. The
maintenance of a correct ovarian function is undoubtedly related to
the presence of a wide vascular bed that is able to sustain
follicular and corpora lutea development. VEGF is the most relevant
among angiogenetic signals. Its expression is regulated by many
different signals. Hypoxia acts by HIF-1(Hypoxia-inducible factor
1) transcription factor induction and represents one of the most
efficient stimuli together with vasoactive gaseous stimulus as CO e
NO. The evaluation of the expression of HIF-1 as well as of HO (-1
and -2) and NOS enzymes, responsible of CO e NO production
respectively, in relation to functional stage of ovarian structures
are investigated.
2- IN VIVO, EX VIVO AND IN VITRO ANIMAL MODELS FOR
INFLAMMATION STUDIES IN SWINE SPECIES
Experimental approaches:
Experience on swine animal model in acute or chronic trials
(endotoxic shock, gene terapy, ex vixo studies, ecc);
experimental surgery on swine of different ages; derivation of
primary cell cultures from animal tissue (endothelial cell from
aorta or specific tissues microvasculature); study of physiological
systems involved in pathologies development, mainly inflammation;
assessment of animal models for human diseases; genomics and
proteomics approaches specific for swine species.
Principal fields of research:
The pig is worldwide recognized as the most important preclinical
model on the bases of its anatomical and physiological similarity
with humans. Organs dimensions, digestive physiology, renal
physiology, pulmonary vascular structure and physiology, coronary
distribution, respiratory indexes, cardiovascular anatomy and
physiology. Thereafter, in the “Animal Models” context the swine
give a wide set of non-infective syndromes useful for studying
analogue human syndromes. Endothelial cells are fundamental in
inflammatory response organization and propagation and regulate
vasoconstriction process, leucocytes adhesion, coagulation and
thrombosis. These modifications are mainly due to the expression of
pro-inflammatory genes in endothelial cells, coding for adhesion
molecules, cytokines, chemokines, etc. the expression of so-called
“protective genes” e.g. (HO-1), A20, NOS modulate the inflammatory
answer.
3- TRANSGENIC PIG PRODUCTION
Experimental approaches:
Production of transgenic and multi-transgenic pig for biomedical
purposes, characterization and breeding; derivation of primary cell
cultures from swine tissue (endothelial cell from aorta or specific
tissues microvasculature; fibroblasts from skin or internal
organs); in vitro production of swine embryos; ethological and
welfare assessment in pigs; genomics and proteomics approaches
specific for swine species.
Principal fields of research:
To prevent Hyperacute Rejection following the xenotransplantation,
we produced hDAF transgenic pigs by means of Sperm Mediated Gene
Transfer technology. The validity of experimental technology was
confirmed by ex vivo perfusion of organs. New approaches are needed
to overcome the acute vascular rejection that is based on a
generalized pro-inflammatory state of endothelial cells and
monocytes. Protective genes as A20, able to counteract the
activation of NF-kB and HO-1 could be introduced constitutively in
transgenic pigs. The multitransgenic pig production is a main goal
on xenotransplatation studies but is also a very interesting tool
for basic and applied research. Moreover the application of SMGT to
gene silencing by siRNA is a very interesting improvement of the
technique.
