Localization of the glucocorticoid receptor in rat brain mitochondria.

The distribution of glucocorticoid receptor in subcellular fractions of brain cortex and hippocampus, two regions rich in glucocorticoid receptor, has revealed its presence in nuclei, cytosol, mitochondria, synaptosomes, and synaptosomal mitochondria. The identification of glucocorticoid receptor has been accomplished both by Western blotting using antibodies recognizing the carboxy and the amino terminus of the glucocorticoid receptor and by immunogold electron microscopy using the same anti-glucocorticoid receptor antibodies. Antibody-glucocorticoid receptor interaction is abolished by preincubation of each antibody with its competing peptide. In addition to the intact 95-kDa glucocorticoid receptor in all fractions, lower molecular weight glucocorticoid receptor fragments have been also detected by Western blotting. The presence of glucocorticoid receptor in brain mitochondria supports the concept of a direct action of glucocorticoids on mitochondrial gene transcription, parallel to the established primary actions of the hormones on nuclear gene transcription, as a mechanism of coordinate regulation of respiratory enzyme biosynthesis by steroid hormones.

Expression of the glucocorticoid receptor in early and late passage C-6 glioma cells and in normal astrocytes derived from aged mouse cerebral hemispheres.

The presence of the glucocorticoid receptor in early and late passage C-6 glioma cells 2B clone and in astrocytes derived from aged mouse cerebral hemispheres has been documented by immunoblotting and/or immunofluorescence labelling. All cell types studied express the glucocorticoid receptor of molecular weight 97 KDa. In addition, in astrocytes derived from aged mouse cerebral hemispheres a smaller molecular weight polypeptide reacting with anti-glucocorticoid receptor antibody was also demonstrated. No difference in the amount of the 97 KDa glucocorticoid receptor between early and late C-6 2B cells was observed, whereas the astrocytes from aged cerebral hemispheres contained considerably reduced amounts of the glucocorticoid receptor compared to C-6 2B cells. Late passage C-6 2B cells were immunofluorescence labelled with the anti-glucocorticoid antibody, the receptor being almost exclusively present in the cytoplasm, with particular concentration in the perinuclear region. The presence of glucocorticoid receptor of molecular weight 97 KDa in glial cells corroborates and expands the existing data based on radioligand binding and immunocytochemical studies. These cell populations can be exploited as a model system for the study of the effects of glucocorticoids on senescence and brain aging.

The identification of type 1 Gaucher disease patients, asymptomatic cases and carriers in The Netherlands using urine samples: an evaluation.

The feasibility of using urine samples for the identification of patients with Gaucher disease and carriers has been investigated. It was found that the pH of a urine sample should be pH 6.0 or lower to ensure stability of lysosomal hydrolases. Two parameters of glucocerebrosidase, which is deficient in Gaucher disease, were studied using urine samples from control subjects, obligate carriers and patients. Firstly, the relative level of glucocerebrosidase activity was measured by relating the activity of the enzyme to that of another lysosomal hydrolase. Secondly, the enzymic activity of glucocerebrosidase per unit of protein was measured using an immunological method. The first method allowed discrimination of nearly all obligate carriers of type 1 Gaucher disease from normal individuals. The second method allowed clear discrimination of the majority of carriers from normal individuals, but some obligate carriers were not distinguishable from normal subjects on the basis of this parameter. However, the combination of both methods allowed discrimination between all obligate carriers examined so far (n = 34) and controls (n = 86). There was variability between healthy individuals in the relative amount of glucocerebrosidase in urine samples. A small proportion of healthy individuals have a relatively high activity of glucocerebrosidase in urine samples, reminiscent of observations made in white blood cells by other investigators. In urine samples from two unrelated parents of Gaucher disease patients a level of glucocerebrosidase activity was present that could not be distinguished from that in samples of patients. These individuals represent cases with subclinical manifestation of Gaucher disease, illustrating once more the remarkable heterogeneity in clinical expression of this disorder.