P03.133 Immunohistochemistry, Immunofluorescence and Confocal Laser Microscopy: An Insight into the Enteric Nervous System During Sheep Scrapie Infection Di Guardo, G1; Marruchella, G1; Ligios, C2; Baffoni, M1; Cancedda, GM2; Macciocu, S2; Gioia, L1; Lalatta Costerbosa, G3; Chiocchetti, R3; Clavenzani, P3; De Grossi, L4; Agrimi, U5; Aguzzi, A6 1Faculty of Veterinary Medicine, University of Teramo, Dept Comparative Biomedical Sciences, Italy; 2Istituto Zooprofilattico Sperimentale della Sardegna, Italy; 3Faculty of Veterinary Medicine, University of Bologna, Dept Vet Morphophysiology and Animal Productions, Italy; 4Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana, Italy; 5Istituto Superiore di Sanità, Dept Food Safety and Veterinary Public Health, Italy; 6University Hospital of Zurich, Institute of Neuropathology, Switzerland The enteric nervous system (ENS) is likely to play a role in the early pathogenesis of sheep scrapie, but little is known about the ENS cell types involved. We investigated the ileal myenteric plexi (MPs) and submucosal plexi (SMPs) of 4 natural and 4 oral experimental scrapie-affected ARQ/ARQ Sarda breed sheep, as well as of 12 healthy sheep carrying different PRNP genotypes. In all control animals, as well as in all scrapie-affected sheep, which were euthanized at the end stage of clinical disease, detailed laboratory investigations were carried out by means of immunohistochemistry (IHC), as well as by indirect immunofluorescence (IIF) and confocal laser microscopy (CLM) techniques. All the above 8 scrapie-affected animals showed IHC evidence of PrPSc deposition within both MPs and SMPs, with IIF and CLM studies allowing us to identify enteroglial cells (EGCs) and, for the first time, also calbindin (CALB)-immunoreactive (IR) and neuronal nitric oxide synthase (nNOS)-IR neurons as the ENS cytotypes involved in PrPSc accumulation and plausibly, thereby, in the subsequent process of “neuroinvasion”. In conclusion, IHC, IIF and CLM proved to be in our hands three highly valuable and complementary laboratory techniques for investigating the p
DI GUARDO G., MARRUCHELLA G., LIGIOS C., BAFFONI M., CANCEDDA G.M., MACCIOCU S., et al. (2007). Immunohistochemistry, Immunofluorescence and Confocal Laser Microscopy: an Insight into the Enteric Nervous System During Sheep Infection.. EDIMBURGH : s.n.
Immunohistochemistry, Immunofluorescence and Confocal Laser Microscopy: an Insight into the Enteric Nervous System During Sheep Infection.
LALATTA COSTERBOSA, GIOVANNA;CHIOCCHETTI, ROBERTO;CLAVENZANI, PAOLO;
2007
Abstract
P03.133 Immunohistochemistry, Immunofluorescence and Confocal Laser Microscopy: An Insight into the Enteric Nervous System During Sheep Scrapie Infection Di Guardo, G1; Marruchella, G1; Ligios, C2; Baffoni, M1; Cancedda, GM2; Macciocu, S2; Gioia, L1; Lalatta Costerbosa, G3; Chiocchetti, R3; Clavenzani, P3; De Grossi, L4; Agrimi, U5; Aguzzi, A6 1Faculty of Veterinary Medicine, University of Teramo, Dept Comparative Biomedical Sciences, Italy; 2Istituto Zooprofilattico Sperimentale della Sardegna, Italy; 3Faculty of Veterinary Medicine, University of Bologna, Dept Vet Morphophysiology and Animal Productions, Italy; 4Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana, Italy; 5Istituto Superiore di Sanità, Dept Food Safety and Veterinary Public Health, Italy; 6University Hospital of Zurich, Institute of Neuropathology, Switzerland The enteric nervous system (ENS) is likely to play a role in the early pathogenesis of sheep scrapie, but little is known about the ENS cell types involved. We investigated the ileal myenteric plexi (MPs) and submucosal plexi (SMPs) of 4 natural and 4 oral experimental scrapie-affected ARQ/ARQ Sarda breed sheep, as well as of 12 healthy sheep carrying different PRNP genotypes. In all control animals, as well as in all scrapie-affected sheep, which were euthanized at the end stage of clinical disease, detailed laboratory investigations were carried out by means of immunohistochemistry (IHC), as well as by indirect immunofluorescence (IIF) and confocal laser microscopy (CLM) techniques. All the above 8 scrapie-affected animals showed IHC evidence of PrPSc deposition within both MPs and SMPs, with IIF and CLM studies allowing us to identify enteroglial cells (EGCs) and, for the first time, also calbindin (CALB)-immunoreactive (IR) and neuronal nitric oxide synthase (nNOS)-IR neurons as the ENS cytotypes involved in PrPSc accumulation and plausibly, thereby, in the subsequent process of “neuroinvasion”. In conclusion, IHC, IIF and CLM proved to be in our hands three highly valuable and complementary laboratory techniques for investigating the pI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
