Selective regulation of growth factor expression in cultured cortical astrocytes by neuro-pathological toxins.

Astrocytes are integrated in the complex regulation of neurodegeneration and neuronal damage in the CNS. It is well-known that astroglia produces a plethora of growth factors which might be protective for neurons. Growth factors prevent neurons from cell death and promote proliferation and differentiation of precursor cells. Previous data suggest that astrocytes may respond to toxic stimuli by a selective mobilization of guarding molecules. In the present study, we have investigated the potency of different pathological stimuli such as lipopolysaccharides, tumor necrosis factor alpha, glutamate, and hydrogen peroxide to activate cultured cortical astroglia and stimulate growth factor expression. Astroglial cultures were exposed to the above factors for 24h at non-toxic concentrations for astrocytes. Growth factor expression was analyzed by real-time PCR, oligo-microarray technique, and ELISA. Insulin-like growth factor-1 was selectively down-regulated by lipopolysaccharides and tumor necrosis factor alpha, bone morphogenetic protein 6 by all stimuli. In contrast, lipopolysaccharides, tumor necrosis factor alpha, and glutamate increased leukemia inhibitory factor. Fibroblast growth factor 2 was up-regulated by lipopolysaccharides and tumor necrosis factor alpha and down-regulated by hydrogen peroxide. Besides hydrogen peroxide, all other stimuli promoted vascular epithelial growth factor A mRNA and protein expression. It appears that lipopolysaccharides but not tumor necrosis factor alpha effects on vascular epithelial growth factor A depend on the classic NFkappaB pathway. Our data clearly demonstrate that astroglia actively responses to diverse pathological compounds by a selective expression pattern of growth factors. These findings make astrocytes likely candidates to participate in disease-specific characteristics of neuronal support or damage.

17beta-estradiol and progesterone prevent cuprizone provoked demyelination of corpus callosum in male mice.

Sex hormones, for example, estrogen and progesterone, are thought to affect and delay progression of multiple sclerosis (MS) in pregnant women. Although both steroid hormones are neuroprotective in the brain and elevated during pregnancy, only estrogen was tested in clinical trials. To evaluate the role of 17beta-estradiol (E) and progesterone (P) in prevention demyelination, young adult male mice were fed with cuprizone for a defined time interval and simultaneously treated with steroids by repeated injections into the neck region. The status of myelination was analyzed by magnetic resonance imaging and conventional histological staining. The individual application of E and P resulted only in a moderate prevention of demyelination in the corpus callosum (CC). The combined treatment with both steroid hormones counteracted the process of demyelination. Expression of the mature (PLP and MBP) and premature (PDGF-alpha-R) oligodendrocyte markers were significantly increased after hormone application in the affected CC. In addition, both hormones stimulated astrogliosis and the expression of IGF-1. Microglial invasion in demyelinated CC was pronounced and additionally localized in the midline of CC after hormone treatment. These data show that sex steroids can protect the brain from demyelination and stimulate remyelination. It appears that only the administration of both hormones is fully effective. The beneficial steroid effect requires interactions with oligodendrocytes possibly by preventing their degeneration or recruitment from precursor cells which are stimulated to remyelinated fibers. The positive hormonal influence on myelination in the CNS may be a future therapeutically strategy for the treatment of MS.

Cuprizone treatment induces demyelination and astrocytosis in the mouse hippocampus.

Memory impairment is outstanding within the spectrum of cognitive deficits in multiple sclerosis (MS) patients. Demyelination has been reported in the hippocampus formation of MS patients. The degree of hippocampus lesions in MS strongly correlates with progression of cognitive dysfunction. Because no appropriate animal model for the study of hippocampus demyelination has been established, we used the cuprizone mouse model to investigated demyelination in young adult and aged mice. The myelin status was analyzed by classical histological staining, immunocytochemistry for proteolipoprotein, and electron microscopy. Oligodendrocyte, astroglial, and microglia markers were studied. Cuprizone intoxication induced an almost complete demyelination of distinct hippocampus subregions to a similar extent in young adult and aged male mice. Demyelination was pronounced in a subset of white and gray matter areas, i.e., the stratum lacunosum moleculare containing the perforant path, medial alveus, stratum pyramidale in the cornu ammonis 2/3 region, and hilus region. Besides demyelination, affected areas displayed hypertrophic and hyperplastic astrocytosis. No significant effect on microglia invasion was detected at any investigated time point (0, 3, 5, and 7 weeks). We conclude that cuprizone-induced demyelination provides an adequate animal model to investigate appropriate therapy strategies for the prevention of hippocampus demyelination.

Brain-region-specific astroglial responses in vitro after LPS exposure.

Astroglia is well-known to be integrated in the complex regulation of neuroinflammation in the central nervous system. Astrocytes become activated and synthesize cytokines, chemokines, and prostanoids during degenerative and vulnerable processes and interact with other immune-competent cells. Degenerative disorders often occur in a brain-region-specific fashion suggesting differences in the activity and reactivity of innate immune cells. We have investigated the potency of lipopolysaccharides (LPS) to differently stimulate astrocytes from the cortex and midbrain. Astroglial cultures were prepared from Bagg albino/c mice and exposed to LPS. Astrocytes from both brain areas already differed in their capacity and profile of cytokine expression under basal unstimulated conditions. In response to LPS, we observed both a region-specific pattern of up-regulation of distinct cytokines and differences in the extent and time-course of activation. Our data demonstrate that astrocytes reveal a region-specific basal profile of cytokine expression and a selective area-specific regulation of cytokines upon LPS-induced inflammation. This makes astrocytes likely candidates to be responsible for region-specific incidence rates of neurological and neurodegenerative disorders.