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Development of the first oligonucleotide microarray for global gene expression profiling in guinea pigs: defining the transcription signature of infectious diseases.

Jain R, Dey B, Tyagi AK - BMC Genomics (2012)

Bottom Line: To validate and demonstrate the merit of this microarray, we have studied, as an example, the expression profile of guinea pig lungs during the advanced phase of M. tuberculosis infection.A significant upregulation of 1344 genes and a marked down regulation of 1856 genes in the lungs identified a disease signature of pulmonary tuberculosis infection.An important gap in the area of infectious diseases has been addressed and a valuable molecular tool is provided to optimally harness the potential of guinea pig model to develop better vaccines and therapies against human diseases.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India.

ABSTRACT

Background: The Guinea pig (Cavia porcellus) is one of the most extensively used animal models to study infectious diseases. However, despite its tremendous contribution towards understanding the establishment, progression and control of a number of diseases in general and tuberculosis in particular, the lack of fully annotated guinea pig genome sequence as well as appropriate molecular reagents has severely hampered detailed genetic and immunological analysis in this animal model.

Results: By employing the cross-species hybridization technique, we have developed an oligonucleotide microarray with 44,000 features assembled from different mammalian species, which to the best of our knowledge is the first attempt to employ microarray to study the global gene expression profile in guinea pigs. To validate and demonstrate the merit of this microarray, we have studied, as an example, the expression profile of guinea pig lungs during the advanced phase of M. tuberculosis infection. A significant upregulation of 1344 genes and a marked down regulation of 1856 genes in the lungs identified a disease signature of pulmonary tuberculosis infection.

Conclusion: We report the development of first comprehensive microarray for studying the global gene expression profile in guinea pigs and validation of its usefulness with tuberculosis as a case study. An important gap in the area of infectious diseases has been addressed and a valuable molecular tool is provided to optimally harness the potential of guinea pig model to develop better vaccines and therapies against human diseases.

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Pulmonary pathology in M. tuberculosis infected guinea pigs. The figure depicts (A) representative photograph showing gross pathology of lungs of M. tuberculosis infected guinea pigs at 10 weeks post-infection; (B) lower magnification photomicrograph of H&E stained, formalin fixed and paraffin embedded 5 μm section of lung tissue exhibiting multiple coalescing granulomas with central necrotic core and (C) photomicrograph (at 40X) of Van Gieson stained 5 μm lung section exhibiting extensive fibrosis as evident from thick bands of collagen (red color) around the granulomatous regions. Scale bar represents 1000 μm.
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Figure 2: Pulmonary pathology in M. tuberculosis infected guinea pigs. The figure depicts (A) representative photograph showing gross pathology of lungs of M. tuberculosis infected guinea pigs at 10 weeks post-infection; (B) lower magnification photomicrograph of H&E stained, formalin fixed and paraffin embedded 5 μm section of lung tissue exhibiting multiple coalescing granulomas with central necrotic core and (C) photomicrograph (at 40X) of Van Gieson stained 5 μm lung section exhibiting extensive fibrosis as evident from thick bands of collagen (red color) around the granulomatous regions. Scale bar represents 1000 μm.

Mentions: For gene expression analysis, RNA was isolated from lung tissues obtained from guinea pigs at 10 weeks following infection with ~500 tubercle bacilli. CFU analysis of guinea pig lungs at this time point revealed the presence of ~6.0 log10 tubercle bacilli/g of tissue. Moreover, infected guinea pigs exhibited severe pathological damage in lungs characterized by numerous large caseating granulomas encompassing central necrosis with occasional coalescence of multiple tubercles (Figure2 A, B). In addition, extensive areas of irregular thick bands of fibrous collagen deposition were observed around the granulomas (Figure2C). Hence, the gene expression in this phase of infection represents a transcription signature of advanced progressive TB disease.


Development of the first oligonucleotide microarray for global gene expression profiling in guinea pigs: defining the transcription signature of infectious diseases.

Jain R, Dey B, Tyagi AK - BMC Genomics (2012)

Pulmonary pathology in M. tuberculosis infected guinea pigs. The figure depicts (A) representative photograph showing gross pathology of lungs of M. tuberculosis infected guinea pigs at 10 weeks post-infection; (B) lower magnification photomicrograph of H&E stained, formalin fixed and paraffin embedded 5 μm section of lung tissue exhibiting multiple coalescing granulomas with central necrotic core and (C) photomicrograph (at 40X) of Van Gieson stained 5 μm lung section exhibiting extensive fibrosis as evident from thick bands of collagen (red color) around the granulomatous regions. Scale bar represents 1000 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475082&req=5

Figure 2: Pulmonary pathology in M. tuberculosis infected guinea pigs. The figure depicts (A) representative photograph showing gross pathology of lungs of M. tuberculosis infected guinea pigs at 10 weeks post-infection; (B) lower magnification photomicrograph of H&E stained, formalin fixed and paraffin embedded 5 μm section of lung tissue exhibiting multiple coalescing granulomas with central necrotic core and (C) photomicrograph (at 40X) of Van Gieson stained 5 μm lung section exhibiting extensive fibrosis as evident from thick bands of collagen (red color) around the granulomatous regions. Scale bar represents 1000 μm.
Mentions: For gene expression analysis, RNA was isolated from lung tissues obtained from guinea pigs at 10 weeks following infection with ~500 tubercle bacilli. CFU analysis of guinea pig lungs at this time point revealed the presence of ~6.0 log10 tubercle bacilli/g of tissue. Moreover, infected guinea pigs exhibited severe pathological damage in lungs characterized by numerous large caseating granulomas encompassing central necrosis with occasional coalescence of multiple tubercles (Figure2 A, B). In addition, extensive areas of irregular thick bands of fibrous collagen deposition were observed around the granulomas (Figure2C). Hence, the gene expression in this phase of infection represents a transcription signature of advanced progressive TB disease.

Bottom Line: To validate and demonstrate the merit of this microarray, we have studied, as an example, the expression profile of guinea pig lungs during the advanced phase of M. tuberculosis infection.A significant upregulation of 1344 genes and a marked down regulation of 1856 genes in the lungs identified a disease signature of pulmonary tuberculosis infection.An important gap in the area of infectious diseases has been addressed and a valuable molecular tool is provided to optimally harness the potential of guinea pig model to develop better vaccines and therapies against human diseases.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India.

ABSTRACT

Background: The Guinea pig (Cavia porcellus) is one of the most extensively used animal models to study infectious diseases. However, despite its tremendous contribution towards understanding the establishment, progression and control of a number of diseases in general and tuberculosis in particular, the lack of fully annotated guinea pig genome sequence as well as appropriate molecular reagents has severely hampered detailed genetic and immunological analysis in this animal model.

Results: By employing the cross-species hybridization technique, we have developed an oligonucleotide microarray with 44,000 features assembled from different mammalian species, which to the best of our knowledge is the first attempt to employ microarray to study the global gene expression profile in guinea pigs. To validate and demonstrate the merit of this microarray, we have studied, as an example, the expression profile of guinea pig lungs during the advanced phase of M. tuberculosis infection. A significant upregulation of 1344 genes and a marked down regulation of 1856 genes in the lungs identified a disease signature of pulmonary tuberculosis infection.

Conclusion: We report the development of first comprehensive microarray for studying the global gene expression profile in guinea pigs and validation of its usefulness with tuberculosis as a case study. An important gap in the area of infectious diseases has been addressed and a valuable molecular tool is provided to optimally harness the potential of guinea pig model to develop better vaccines and therapies against human diseases.

Show MeSH
Related in: MedlinePlus