A Time-Course Study of Gene Responses in Chicken Granulosa Cells Infected by Salmonella Enteritidis
Date Issued
2009
Date
2009
Author(s)
Wang, Chia-Lan Nicole
Abstract
Consumption of Salmonella Enteritidis (SE)-contaminated eggs has been recognized as one of the important reasons of human food-borne salmonellosis. Chicken granulosa cells (CGC), last tissue layer surrounding the yolk in prevulatory follicles has been demonstrated a preferred site for the Salmonella Enteritidis invasion. To understand how the chicken granulosa cell responds to the infection, a time-course in-vitro study was conducted to identify transcription changes of chicken granulosa cell using chicken whole genome microarrays. The expression of 214 genes of chicken granulosa cell had altered (p<0.01) when compared to those of uninfected cells after the infection. According to the different responses of the altered gene between 4hpi and 48hpi, they are divided into cluster 1a, 1b, 1c, 2a, and 2b those include 70 genes (32.71%), 27 genes (12.62%), 66 genes (30.84%), 41 genes (19.16%), and 10 genes (4.67%) respectively. Among of them, many were related to immune response, physiological process, signal transduction activity, transcription and some of unknown function. We also noted that the JAK-STAT pathway which is essential in regulation of cellular cytokines and growth factors being highly active those are IL-6, cytokineR, STAT,and PI3K in the upstream of this pathway. Among the up-regulated genes, the mRNA levels of TLR15, IL-6, CXCLi1, CXCLi2 and K203 at the two time points from the infected cells were also increased by the validation of real-time RT-PCR (RRT-PCR). However the mRNA expression of RASD1 and HB-EGF decreased according to the microarray and RRT-PCR analyses. These results suggest that during the Salmonella Enteritidis infection, chicken granulosa cell are capable of recruiting cells of innate immune responses in short term. In long term, the growth of chicken granulosa cell is suppressed and then the development and ovulation of follicles is also suppressed. Among of them, the chemokine and paracrine play the important role.
Subjects
microarray
chicken granulosa cell
RRTPCR
gene expression
Type
thesis
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