Gene profiling in murine corneas challenged with Aspergillus fumigatus
Yiqiang Wang1, Ting Liu, Huaqing Gong
1State Key Lab Cultivation Base, Shandong Provincial Key Lab of Ophthalmology, Shandong Eye Institute, Qingdao, China.
Purpose:
Fungal keratitis (FK) is a blinding infection of the corneas and accounts for a significant portion of all keratitis in the world but little is known about the pathogenesis of FK, especially at the level of molecular biology. This study tried to determine the genes that are modulated in the process and thus deserve further investigation for understanding the pathogenesis of FK.
Methods:
Agilent Mouse Oligo Microarray was used to compare the gene profiles in control corneas and in corneas challenged with heat-inactivated Aspergillus fumigatus spores. Semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to confirm the changes of interested genes.
Results:
In the 18,335 genes detected by array, 61 showed an increase of more than one-fold in RNA expression and 48 showed a decrease of over 50%. Among the regulated genes some are known to be related to host defense such as IL3, mannose binding lectin A (MBL-A), and prostaglandin D2 synthase (Psgd) while others like Dopachrome tautomerase (Dct) have never been correlated to ocular or host defense. The other 33 changed genes either have unknown functions or encode hypothetical proteins. The changes in the expression of early growth response 4 (Egr4), Dct, Psgd, MBL-A, and hemoglobin alpha adult chain 1 were confirmed by using RT-PCR. MBL-A expression regulation was further confirmed by using immunohistochemistry in both in vitro and in vivo challenged murine corneas.
Conclusions:
MBL-A is among the primary responding genes during the onset of fungal keratitis. Also it was found that the microarray is a useful tool in elucidating the pathogenesis of FK.
Insights
This study identified key genes modulated during fungal keratitis, revealing Mannose-binding lectin A (MBL-A) as a primary responder. Microarray analysis is effective for understanding fungal keratitis pathogenesis.
Area of Science:
- Ophthalmology
- Molecular Biology
- Immunology
Background:
- Fungal keratitis (FK) is a significant cause of blindness globally.
- The molecular pathogenesis of FK remains poorly understood.
- Identifying modulated genes is crucial for understanding FK development.
Purpose of the Study:
- To identify genes modulated during fungal keratitis using microarray analysis.
- To investigate the molecular mechanisms underlying FK pathogenesis.
- To pinpoint genes for further research into FK development.
Main Methods:
- Gene expression profiling using Agilent Mouse Oligo Microarray.
- Comparison of gene profiles in control and Aspergillus fumigatus-challenged corneas.
- Validation of gene expression changes using RT-PCR and immunohistochemistry.
Main Results:
- Out of 18,335 genes, 61 showed increased and 48 decreased RNA expression.
- Key host defense genes like Mannose-binding lectin A (MBL-A) and prostaglandin D2 synthase (Psgd) were modulated.
- Novel genes such as Dopachrome tautomerase (Dct) were identified with potential roles in FK.
- MBL-A expression was confirmed as an early response gene in FK.
Conclusions:
- Mannose-binding lectin A (MBL-A) is a primary responding gene in fungal keratitis.
- Microarray analysis is a valuable tool for elucidating FK pathogenesis.
- Further research into identified genes can enhance understanding of FK.
