No abstract

Phospholipase D (PLD) is a ubiquitous enzyme involved in many important cellular functions. Rat brain PLD isoforms have recently been cloned and characterized, but the expression of this gene has not been well studied. We thus examined the developmental expression of PLD2 in postnatal brains by Northern blotting employing a non-radioactive RNA probe. Two PLD2 mRNA transcripts of 3.9 and 10.8 kb were detected in different brain regions and various tissues of the rat. In postnatal brains, the level of PLD2 mRNA transcripts are low 1 day after birth, increase progressively during development, and reach the maximum level in the adult brain. We thus conclude that rat brain PLD2 mRNA is developmentally regulated.

In the present study the acute effect of intravenous aluminum chloride (1 mg/kg) on choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities of rats was investigated. Aluminum was found to cross the blood-brain barrier (BBB) as indicated by the detection of aluminum in the cerebrospinal fluid (CSF) 30 min after femoral vein injection. Two hours following aluminum injection, ChAT activity in the basal forebrain and hippocampus was significantly reduced by 30% and 22%, respectively, whereas no change was observed in the caudate nuclei. On the other hand, AChE activity was significantly increased by 45% in the caudate nuclei, whereas little change was observed in other brain areas. This report demonstrates that rapid transport of Al across the BBB, and the acute nature of Al neurotoxicity in rats.

In this study, alpha (alpha) isoform proteins were purified from the partially purified Na,K-ATPase by SDS-PAGE and electroelution. Peptide mapping showed subtle biochemical differences between alpha subunit proteins of rat and human origin. The purified alpha proteins were treated with formic acid, the cleaved polypeptide fragments were separated by SDS-PAGE, the bands corresponding to 40, 50, and 60 kDa were excised, and the proteins were electroeluted. The purified 40, 50, and 60 kDa polypeptides were essentially homogeneous, and were used for preparation of polyclonal antibodies in rabbits. The antisera to alpha proteins (R alpha) and 60 & 40 kDa polypeptides (R60 & R40) were obtained and characterized by Western blotting. All three antisera were highly specific, since they cross-reacted with only the 100 kDa bands of the crude brainstem homogenates, of the axolemma, and of the cerebral cortex synaptosomes and microsomes. R alpha and R40 were successfully used for immunohistochemical staining of fibers in the white matter of the human brain frontal cortex. These antisera were not isoform-specific, they cross-reacted with 40, 50, and 60 kDa polypeptides as well as the three alpha bands.