Brain fibronectin expression in prenatally irradiated mice.

Abstract

Activation of gene transcription by radiation has been recently demonstrated in vitro. However, little is known on the specificity of these alterations on gene transcription. Prenatal irradiation is a known teratogen that affects the developing mammalian central nervous system (CNS). Altered neuronal migration has been suggested as a mechanism for abnormal development of prenatally irradiated brains. Fibronectin (FN), an extracellular glycoprotein, is essential for neural crest cell migration and neural cell growth. In addition, elevated levels of FN have been found in the extracellular matrix of irradiated lung. To test whether brain FN is affected by radiation, either FN level in insoluble matrix fraction or expression of FN mRNA was examined pre- and postnatally after irradiation. Mice (CD1), at 13 d of gestation (DG), served either as controls or were irradiated with gamma rays at 0.5 or 1 Gy. Control and irradiated animals were killed either at 13 DG, 14 DG, 17 DG, or 5, 6, or 14 d postnatal. Brain and liver were collected from offspring and analyzed for either total FN protein levels or relative mRNAs for FN and tubulin. Results of prenatal irradiation on reduction of postnatal brain weight relative to whole body weight and morphological reduction in cerebral cortex regions of postnatal brains are comparable to that reported by others. Insoluble matrix fraction (IMF) per gram of brain, liver, lung, and heart weight was not significantly different either between control and irradiated groups or between postnatal stages, suggesting that radiation did not affect the IMF. However, total amounts of FN in brain IMF at 17 DG were significantly different (p < .02) between normal (1.66 +/- 0.80 micrograms) and irradiated brains (0.58 +/- 0.22 microgram). FN mRNA was detectable at 13, 14, and 17 DG, but was not detectable at 6 and 14 d postnatal, indicating that FN mRNA is developmentally regulated. After 0.5 Gy of irradiation, expression of FN mRNA was reduced to 36% +/- 22% (1 h), 52% +/- 10% (1 d), and 76% +/- 10% (4 d) of the control level. After 1 Gy of irradiation, relative FN mRNA was 62% +/- 28% (1 h) and 75% +/- 3% (4 days) to the control level, respectively. This reduction was comparable to that reported by others for the cytoskeletal protein beta-actin. In contrast, mRNA for tubulin, another cytoskeletal protein, increased at 1 h after irradiation but then approached normal postnatally. The longer lasting alteration of FN may be more directly related to neural development, particularly if the reduction in FN is nonuniform.