Isolation of cDNAs encoding four mouse FGF family members and characterization of their expression patterns during embryogenesis

JM Hébert, C Basilico, M Goldfarb, O Haub… - Developmental …, 1990 - Elsevier
JM Hébert, C Basilico, M Goldfarb, O Haub, GR Martin
Developmental biology, 1990Elsevier
To initiate a study of the role of the fibroblast growth factor (FGF) family in mammalian
development, we have isolated cDNAs encoding four mouse FGF family members, aFGF,
bFGF, kFGF, and FGF-5. This was achieved by a process that circumvents the use of cDNA
libraries: for each family member, a cDNA fragment containing the conserved portion of the
coding region was amplified from a pool of embryonic and teratocarcinoma cell cDNAs
using the polymerase chain reaction (PCR) and cloned; the remaining coding sequences …
Abstract
To initiate a study of the role of the fibroblast growth factor (FGF) family in mammalian development, we have isolated cDNAs encoding four mouse FGF family members, aFGF, bFGF, kFGF, and FGF-5. This was achieved by a process that circumvents the use of cDNA libraries: for each family member, a cDNA fragment containing the conserved portion of the coding region was amplified from a pool of embryonic and teratocarcinoma cell cDNAs using the polymerase chain reaction (PCR) and cloned; the remaining coding sequences 5′ and 3′ to the conserved region were cloned using the RACE method. The cDNA clones obtained were used as probes to analyze the expression of these genes at the RNA level in teratocarcinoma cells and embryos at 10.5 to 17.5 days of gestation. Fgfk appears to be specific to undifferentiated teratocarcinoma stem cells. Fgf5 transcripts were detected at every stage and in every tissue tested, but showed a dramatic 15-fold increase in abundance as teratocarcinoma stem cells differentiated to simple embryoid bodies. Fgfb expression showed the greatest tissue-specific variability in abundance, with the highest levels detected in the developing limbs and tail. Fgfa showed the least variable pattern of expression, with transcripts detected at roughly equivalent levels in almost all samples analyzed. On the basis of these data we speculate on some possible roles that the different FGF family members may play in the developing embryo.
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