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Goat uterine epithelial cells are susceptible to infection with Caprine Arthritis Encephalitis Virus (CAEV) in vivo.

Ali Al Ahmad MZ, Dubreil L, Chatagnon G, Khayli Z, Theret M, Martignat L, Chebloune Y, Fieni F - Vet. Res. (2012)

Bottom Line: Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence.The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples.The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections.

View Article: PubMed Central - HTML - PubMed

Affiliation: LUNAM University, Oniris, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Sanitary Security of Reproduction Biotechnology Unit, F-44307 Nantes, France. francis.fieni@oniris-nantes.fr.

ABSTRACT
The aim of this study was to determine, using immunofluorescence and in situ hybridization, whether CAEV is capable of infecting goat uterine epithelial cells in vivo. Five CAEV seropositive goats confirmed as infected using double nested polymerase chain reaction (dnPCR) on leucocytes and on vaginal secretions were used as CAEV positive goats. Five CAEV-free goats were used as controls. Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence. The results from dnPCR confirmed the presence of CAEV proviral DNA in the uterine horn samples of infected goats whereas no CAEV proviral DNA was detected in samples taken from the uninfected control goats. The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples. The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections. Finally, laser confocal analysis of double p28/cytokeratin immunolabelled transverse sections of CAEV infected goat uterus, demonstrated that the virus was localized in glandular and epithelial cells. This study clearly demonstrates that goat uterine epithelial cells are susceptible to CAEV infection in vivo. This finding could help to further our understanding of the epidemiology of CAEV, and in particular the possibility of vertical transmission.

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In situ hybridization of uterine tissues (bar = 10 micrometers). CAEV RNA positive staining (brownish-purple intracytoplasmic inclusions) could be identified (black arrow) in the uterine glands ① and in the lamina propria of the mucosa ② of uterine sections.
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Figure 1: In situ hybridization of uterine tissues (bar = 10 micrometers). CAEV RNA positive staining (brownish-purple intracytoplasmic inclusions) could be identified (black arrow) in the uterine glands ① and in the lamina propria of the mucosa ② of uterine sections.

Mentions: Hybridizing cells were characterized by the presence of brownish-purple intracytoplasmic inclusions. The positively-staining cells were concentrated in the mucosa of the lamina propria of uterine sections (Figure 1) and the uterine glands. In the adjacent sections, the glandular epithelial cells could be distinguished by their cuboid morphology with nuclei at the lower end, and were confirmed by brown cytokeratin staining (Figure 2).


Goat uterine epithelial cells are susceptible to infection with Caprine Arthritis Encephalitis Virus (CAEV) in vivo.

Ali Al Ahmad MZ, Dubreil L, Chatagnon G, Khayli Z, Theret M, Martignat L, Chebloune Y, Fieni F - Vet. Res. (2012)

In situ hybridization of uterine tissues (bar = 10 micrometers). CAEV RNA positive staining (brownish-purple intracytoplasmic inclusions) could be identified (black arrow) in the uterine glands ① and in the lamina propria of the mucosa ② of uterine sections.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3298472&req=5

Figure 1: In situ hybridization of uterine tissues (bar = 10 micrometers). CAEV RNA positive staining (brownish-purple intracytoplasmic inclusions) could be identified (black arrow) in the uterine glands ① and in the lamina propria of the mucosa ② of uterine sections.
Mentions: Hybridizing cells were characterized by the presence of brownish-purple intracytoplasmic inclusions. The positively-staining cells were concentrated in the mucosa of the lamina propria of uterine sections (Figure 1) and the uterine glands. In the adjacent sections, the glandular epithelial cells could be distinguished by their cuboid morphology with nuclei at the lower end, and were confirmed by brown cytokeratin staining (Figure 2).

Bottom Line: Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence.The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples.The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections.

View Article: PubMed Central - HTML - PubMed

Affiliation: LUNAM University, Oniris, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Sanitary Security of Reproduction Biotechnology Unit, F-44307 Nantes, France. francis.fieni@oniris-nantes.fr.

ABSTRACT
The aim of this study was to determine, using immunofluorescence and in situ hybridization, whether CAEV is capable of infecting goat uterine epithelial cells in vivo. Five CAEV seropositive goats confirmed as infected using double nested polymerase chain reaction (dnPCR) on leucocytes and on vaginal secretions were used as CAEV positive goats. Five CAEV-free goats were used as controls. Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence. The results from dnPCR confirmed the presence of CAEV proviral DNA in the uterine horn samples of infected goats whereas no CAEV proviral DNA was detected in samples taken from the uninfected control goats. The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples. The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections. Finally, laser confocal analysis of double p28/cytokeratin immunolabelled transverse sections of CAEV infected goat uterus, demonstrated that the virus was localized in glandular and epithelial cells. This study clearly demonstrates that goat uterine epithelial cells are susceptible to CAEV infection in vivo. This finding could help to further our understanding of the epidemiology of CAEV, and in particular the possibility of vertical transmission.

Show MeSH
Related in: MedlinePlus