We investigated the ileal myenteric (MPs) and submucosal plexuses (SMPs) of 32 Sarda breed sheep carrying

different PrP genotypes (ARQ/ARQ, ARQ/AHQ, ARQ/ARR, ARR/ARR), which had been orally dosed with

scrapie at 8 months of age and euthanized at definite time intervals post-infection (p.i.)

Agrimi, Umberto

Albanese, Valeria

Chiocchetti, Roberto

Clavenzani, Paolo

D'Agostino, Claudia

De Grossi, Luigi

Di Guardo, Giovanni

Giordani, Francesco

Lalatta-Costerbosa, Giovanna

Ligios, Ciriaco

Malatesta, Daniela

Marruchella, Giuseppe

Mazzoni, Maurizio

Nonno, Romolo

Petrucci, Luigi

Rosone, Francesca

Sarli, Giuseppe

Vaccari, Gabriele

Italian

UnissResearch

 AIPVet 
 
Associazione Italiana di Patologia Veterinaria 
 
 
ATTI III Congresso Nazionale 
 
ISSN 1825-2265 
 
 
 
 
con la partecipazione 
 
della Società Italiana di Patologia Tossicologica e Sperimentale 
 
del Gruppo di Patologia Clinica Veterinaria 
 
 
 
 
Pisa, 11-13 Maggio 2006 
 
 
Associazione Italiana di Patologia Veterinaria – Italian Association of Veterinary Pathologists 
www.aipvet.it  
 
III Convegno Nazionale AIPVet 
Comunicazioni 
- 26 - 
 
INDAGINI PATOGENETICHE SUI PLESSI DEL SISTEMA NERVOSO ENTERICO DI 
OVINI DI RAZZA SARDA INFETTATI PER OS CON L’AGENTE DELLA SCRAPIE E CON 
DIVERSA SUSCETTIBILITA’ GENETICA NEI CONFRONTI DELLA MALATTIA* 
 
Marruchella Giuseppe1, Malatesta Daniela1, Petrucci Luigi 1, Lalatta Costerbosa Giovanna 2, 
Clavenzani Paolo 2, Chiocchetti Roberto2, Mazzoni Maurizio2, Albanese Valeria2, Agrimi 
Umberto3, D’Agostino Claudia3, Vaccari Gabriele3, Nonno Romolo3, De Grossi Luigi 4, Rosone 
Francesca4, Giordani Francesco4, Ligios Ciriaco5, Sarli Giuseppe6,  Di Guardo Giovanni 1 
 
1Università degli Studi di Teramo, Facoltà di Medicina Veterinaria, Dipartimento di Scienze 
Biomediche Comparate, Teramo; 2Università degli Studi di Bologna, Facoltà di Medicina Veterinaria, 
Dipartimento di Morfofisiologia Veterinaria e Produzioni Animali, Ozzano Emilia (Bologna); 3Istituto 
Superiore di Sanità, Dipartimento di Sanità Alimentare e Animale, Roma; 4Istituto Zooprofilattico 
Sperimentale delle Regioni Lazio e Toscana, Viterbo; 5Istituto Zooprofilattico Sperimentale della 
Sardegna, Sassari; 6Università degli Studi di Bologna, Facoltà di Medicina Veterinaria, Dipartimento 
di Sanità Pubblica Veterinaria e Patologia Animale, Ozzano Emilia (Bologna). 
 
Sebbene il sistema nervoso enterico (ENS) svolga un ruolo cruciale nella scrapie ovina, non vi sono dati sulle 
popolazioni cellulari coinvolte nell’infezione, né sugli eventuali danni da esse subiti. 
Il presente studio è stato condotto sui plessi mienterici e sottomucosi dell’ileo di 32 pecore di razza Sarda, 
recanti diversi genotipi (ARQ/ARQ, ARQ/AHQ, ARQ/ARR, ARR/ARR) con riferimento al gene codificante per 
la PrPC. I suddetti animali, infettati per os all’età di 8 mesi con un ceppo di scrapie caratterizzato nel topo, sono 
stati sacrificati mediante eutanasia a determinati intervalli di tempo dall’infezione (p.i.). E’ stata quindi valutata, 
tramite immunoistochimica ed immunofluorescenza indiretta su sezioni tessutali e su preparati “wholemount”, 
l’immunoreattività (IR) nei confronti della PrPSc, del “marker” pan-neuronale Hu C/D, dell’ossido-nitrico 
sintetasi (nNOS), della calbindina (CALB), della proteina fibrillare acida gliale (GFAP) e della sinaptofisina. 
In 8 pecore con genotipo ARQ/ARQ, clinicamente sane e sacrificate a 12-24 mesi p.i., nonché in 4 ovini 
clinicamente affetti (1 con genotipo ARQ/ARQ, 3 con genotipo ARQ/AHQ), questi ultimi sacrificati 
rispettivamente a 24, 35, 36,3 e 39,5 mesi p.i., le indagini immunoistochimiche hanno rivelato la presenza di 
PrPSc sia nell’encefalo (obex), sia nell’ENS, specialmente nei plessi mienterici. In tali distretti il deposito della 
PrPSc era compatibile con un interessamento delle cellule enterogliali (EGCs), mentre in 6 animali con genotipo 
ARQ/ARQ era evidente un drastico calo numerico (P < 0,05) dei neuroni CALB-IR rispetto ai soggetti di 
controllo ed a quelli con differente genotipo infettati per os. Inoltre, i neuroni CALB-IR mostravano una 
pressoché esclusiva IR nucleare. 
I nostri dati supportano un coinvolgimento delle EGCs e dei neuroni CALB-IR dei plessi nervosi ileali nella 
scrapie ovina.   
 
Parole chiave: Scrapie sperimentale, sistema nervoso enterico, ovino 
 
 
PATHOGENETIC INVESTIGATIONS ON THE ENTERIC NERVOUS SYSTEM PLEXUSES 
OF SARDA BREED SHEEP WITH DIFFERENT PrP GENOTYPES FOLLOWING ORAL 
EXPERIMENTAL SCRAPIE INFECTION 
 
The enteric nervous system (ENS) plays a key role in sheep scrapie, but no information exists on the cytotypes 
which are involved during infection, nor on the damage of such cells. 
We investigated the ileal myenteric (MPs) and submucosal plexuses (SMPs) of 32 Sarda breed sheep carrying 
different PrP genotypes (ARQ/ARQ, ARQ/AHQ, ARQ/ARR, ARR/ARR), which had been orally dosed with 
scrapie at 8 months of age and euthanized at definite time intervals post-infection (p.i.). PrPSc immunoreactivity 
(IR), along with neuronal marker Hu C/D, nitric oxide synthase (nNOS), calbindin (CALB), glial fibrillary acidic 
protein (GFAP) and synaptophysin IR, were evaluated by immunohistochemistry (IHC) and indirect 
immunofluorescence on paraffin sections and wholemount preparations. 
Eight clinically-healthy ARQ/ARQ sheep euthanized at 12-24 months p.i., along with 1 ARQ/ARQ (euthanized 
at 24 months p.i.) and 3 ARQ/AHQ (euthanized at 35, 36.3 and 39.5 months p.i., respectively) clinically-affected 
sheep, showed IHC evidence of PrPSc in both their brain (obex) and ENS, especially in MPs. PrPSc deposition 
 
 
Associazione Italiana di Patologia Veterinaria – Italian Association of Veterinary Pathologists 
www.aipvet.it  
 
III Convegno Nazionale AIPVet 
Comunicazioni 
- 27 - 
was compatible with an involvement of enteroglial cells (EGCs) and a dramatic reduction (P value < 0.05) of 
CALB-IR neurons was observed in the MPs of 6 ARQ/ARQ animals, as compared to normal healthy controls 
and to infected subjects of different PrP genotypes, which also showed an almost exclusive nuclear IR of these 
cells. 
Therefore, EGCs and CALB-IR neurons of ileal ENS plexuses are likely involved in sheep scrapie. 
 
Keywords: Experimental scrapie, enteric nervous system, sheep 
 


Pathogenetic investigations on the enteric nervous system plexuses of sarda breed sheep with different PrP genotypes following oral experimental scrapie infection

The enteric nervous system (ENS) plays a key role in sheep scrapie, but no information exists on the cytotypes

which are involved during infection, nor on the damage of such cells.

We investigated the ileal myenteric (MPs) and submucosal plexuses (SMPs) of 32 Sarda breed sheep carrying

different PrP genotypes (ARQ/ARQ, ARQ/AHQ, ARQ/ARR, ARR/ARR), which had been orally dosed with

scrapie at 8 months of age and euthanized at definite time intervals post-infection (p.i.). PrPSc immunoreactivity

(IR), along with neuronal marker Hu C/D, nitric oxide synthase (nNOS), calbindin (CALB), glial fibrillary acidic

protein (GFAP) and synaptophysin IR, were evaluated by immunohistochemistry (IHC) and indirect

immunofluorescence on paraffin sections and wholemount preparations.

Eight clinically-healthy ARQ/ARQ sheep euthanized at 12-24 months p.i., along with 1 ARQ/ARQ (euthanized

at 24 months p.i.) and 3 ARQ/AHQ (euthanized at 35, 36.3 and 39.5 months p.i., respectively) clinically-affected

sheep, showed IHC evidence of PrPSc in both their brain (obex) and ENS, especially in MPs. PrPSc depositino

was compatible with an involvement of enteroglial cells (EGCs) and a dramatic reduction (P value &lt; 0.05) of

CALB-IR neurons was observed in the MPs of 6 ARQ/ARQ animals, as compared to normal healthy controls

and to infected subjects of different PrP genotypes, which also showed an almost exclusive nuclear IR of these

cells.

Therefore, EGCs and CALB-IR neurons of ileal ENS plexuses are likely involved in sheep scrapie

Pathogenetic investigations on the enteric nervous system plexuses of Sarda breed sheep with different PrP genotypes following oral experimental scrapie infection

Pathogenetic investigations on the enteric nervous system plexuses of sarda breed sheep with different PrP genotypes following oral experimental scrapie infection

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