Spatial and temporal expression of P-glycoprotein in the congenitally hydrocephalic HTX rat brain.

ABSTRACT

The purpose of the present study is to examine spatial and temporal expression of P-glycoprotein in the brain of congenitally hydrocephalic HTX rats. P-glycoprotein has been reported not only as a drug efflux pump but also one of the factors that restricts brain edema. We examined the rat brain from postnatal day 1 to 60 using light and electron microscopy, immunohistochemistry, Western immunoblot and reverse transcriptase-polymerase chain reaction (RT-PCR) methods with monoclonal antibody specific for P-glycoprotein. Immunohistochemically, the positive anti-P-glycoprotein reactivity was found in capillaries of the normal control rat cerebrum. In the hydrocephalic HTX rat brains, it was also found in the capillaries, but only very weak to no reactivity was found in the capillaries of the spongy changes and cystic wall in the subcortical and lateral periventricular white matter. Immunoelectron microscopically, the reaction product was found exclusively on the luminal surface of the capillary endothelium in control rats. A tracer study with intracardiac perfusion of lanthanum chloride showed that lanthanum penetrated the tight junctions and passed through the intercellular space. In the Western immunoblot analysis, P-glycoprotein of 170 kDa was detected clearly in most normal control rat brains but it was not found in the hydrocephalic HTX rat brains. Moreover, mdr1 P-glycoprotein gene expression in the subcortical white matter was examined by RT-PCR. It was detected in all normal control rat brains, but not found in the hydrocephalic HTX rat brains. The results suggested that the absence of P-glycoprotein expression in the capillaries of deep subcortical and lateral periventricular white matter of hydrocephalic HTX rats led to a deficiency of the blood-brain barrier and might be related to vasogenic edema and to the formation of the spongy changes and cystic cavities.