The effects of hypoxia on growth cones in the ovine fetal brain.

OBJECTIVE

This study was designed to investigate the effects of hypoxia on neural process proliferation by studying its effects on growth cone tubulin and insulin-like growth factor (IGF)-I receptor content.

METHODS

Six fetal lambs were catheterized in the brachial artery and vein. Maternal oxygenation was reduced in steps from a fractional inspired oxygen concentration (FiO2) of 20% to 6% by addition of nitrogen to the inhaled gas mixture for a period of 4 h of reduced oxygen intake. Fetal arterial blood was sampled after the maternal FiO2 and oxygen were stable for >5 min at maternal FiO2 of 20% to 6%. Controls were obtained from normoxic fetuses whose ewes had similar surgery and were kept at an FiO2 of 20% throughout the experiment. Growth cones were isolated from the fetal cerebrum and cerebellum. alpha-tubulin and IGF-I receptors were quantified by immunoblotting. Tubulin and IGF-I receptor mRNA expressions were quantified by real-time polymerase chain reaction.

RESULTS

Maternal nitrogen breathing reduced fetal arterial pH from 7.32+/-0.06 to 6.99+/-0.02 (p<0.001). Hypoxia increased IGF-I receptors from 143+/-10 to 327+/-14 (p<0.001) and from 272+/-26 to 396+/-34 (p<0.001) fluorescence units/microg protein in the cerebrum and cerebellum, respectively. It also increased alpha-tubulin from 713+/-30 to 1873+/-126 (p<0.001) and from 780+/-34 to 2362+/-79 (p<0.001) fluorescence units/microg protein in the cerebrum and cerebellum, respectively. Expression of IGF-I receptor mRNA increased significantly in the hypoxic animals both in the cerebrum and the cerebellum, but there was no change in expression of alpha-tubulin mRNA.

CONCLUSIONS

This increase in IGF-I receptor expression and growth cone content may be an adaptive response to hypoxia to maintain neurite growth by facilitating binding of IGF-I. Hypoxia also increased the growth cone level of alpha-tubulin but did not increase its mRNA expression, which may indicate an inability to polymerize tubulin and build microtubules.