Transcriptional regulation of GABAA receptor gamma2 subunit gene.

We have cloned the promoter regions of the genes for the mouse and human gamma2 subunits of the type A receptors for gamma-aminobutyric acid (GABA). For the mouse, the two major transcription start sites were at +1 (by definition) and +43, as established by rapid amplification of cDNA ends (RACE) and primer extension. This numbering places the start methionine at +297. There was no TATA or CCAAT box. Both mouse and human sequences have a candidate neuron-restrictive silencer element (NRSE) site in the first intron (+956 in mouse). We made assorted mouse-based promoter/reporter (luciferase) constructs starting from a core extending from -331 to +136, varying sizes at both ends, and including and excluding the putative NRSE and more proximal sequences. These were tested by transient transfection in several neuron-like and non-neuronal cell lines. Both proximal and distal downstream elements appeared to help direct expression to neuron-like cells, the NRSE in the intron, by repression in non-neurons, and a 24-bp portion of the 5' untranslated region starting at +113 (named GPE1) by preferentially promoting expression in neuron-like cells. Cotransfected human NRSF (transcription factor for NRSE) reduced reporter expression in neuron-like cells for constructs containing the NRSE in two locations. In gel mobility shift assays, the mouse gamma2 NRSE and a consensus NRSE both bound in vitro translated NRSF very similarly, and the NRSF gave the same major shifted band with the mouse gamma2 NRSE as was observed with nuclear extracts.

[Selenoproteins in rat brain].

Male Wistar rats fed with diets containing eight different levels of selenium(Se). Six rats in each group were killed after 20 weeks to obtain brains. The other 105 rats in the Se depleted group were then divided into four groups randomly and fed with diets containing four different levels of Se. The rats in these four groups were then killed at different time points to observe the kinetic change of selenoproteins. The lowest dietary Se required for reaching the plateau of the activities of cellular glutathione peroxidase (cGPX), phospholipid hydroperoxide glutathione peroxidase (PHGPX) and type II deiodinase (ID II) were 0.05, 0.03 and 0.01 mg/kg respectively. The lowest dietary Se required for reaching normal expression of selenoprotein P and selenoprotein W were 0.01 and 0.05 mg/kg respectively. While the rats were restored Se from diets supplemented with Se, the expression of selenoprotein P and type II deiodinase were in preference to PHGPX and cGPX, and the later two parameters were in preference to selenoprotein W. The results suggested that the function of selenoprotein P and ID II in brain were more important than the other three selenoproteins.

[Selenoproteins in rats with chronic selenium intoxication].

Weaning male Wistar rats were fed with a Torula-yeast based semisynthetic diet supplemented with Na2SeO3 to provide selenium (Se) 0.2 or 0.5 mg/kg (adequate Se or high Sediet) respectively for 20 weeks. By the end of experiment, rats were sacrificed and various tissue of rats were collected to determine the activities of Se-containing enzymes and the mRNAs level of selenoprotein P and selenoprotein W and Se concentration. Livers were examined for pathological changes. It was found that the gain of body weight of the high Se group was significantly lower than that of adequate Se group. Much more Se was accumulated in the tissue of high Se group. The activities of eGPX in plasma, cGPX in kidney, heart and testis, ID I in liver, kidney and thyroid and PHGPX in heart and testis in the high Se group were significantly lower than those in the adequate Se group. However, no specific pathological changes have been found in the liver of both groups. The results suggested that these enzymatic changes could be used as early biochemical parameters for chronic selenium intoxication.