Transcriptional profiling of liver in riboflavin-deficient chicken embryos explains impaired lipid utilization, energy depletion, massive hemorrhaging, and delayed feathering

Table 1 Experimental Design

Fertile eggs, derived from riboflavin binding protein deficient (rd/rd) Single-comb White Leghorn (SCWL) chickens, were injected with riboflavin 5 phosphate (FMN) [riboflavin-rescued (Rf+)] or saline [riboflavin-deficient (Rf-)] at the onset of incubation (e0). Eight viable embryos were killed for liver samples at e9 and e11, while four viable embryos were killed for liver samples at e13 and e15. Two liver samples were randomly pooled to provide 4 biological replicates for each treatment group at e9 and e11, while 4 individual livers samples represent biological replicates for each treatment group at e13 and e15. The very small size of the liver in e9 and e11 required pooling of 2 livers to create each of the 4 biological replicates per riboflavin treatment group for the first two ages (e9 and e11). Total RNA was purified from these 32 liver samples and used for both microarray analysis and qRT-PCR analysis

ISSN: 1471-2164