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CPPG is a highly potent group III metabotropic glutamate receptor (mGluR4/6/7/8) antagonist with selective affi nity to mGluR4. The aim of the present study was to determine the effects of CPPG on spatial memory in the water maze, and on the activity of MMP-2 and MMP9 metalloproteases in the hippocampus of rats without and after hypoxia. Short-term hypoxia (2% O2 and 98% N2) enhances excitatory synaptic transmission and impairs spatial acquisition in the water maze. We observed that CPPG prolonged only escape latency during the fi rst day of training in water maze in group without hypoxia, while it did not infl uence escape latency, distance travelled and swim speed during training of rats after hypoxia. It did not change previously acquired escape response during free-swim trial in either group of rats. Hypoxia signifi cantly increased the activity of the proform of MMP-9 in the hippocampus. Gel zymography showed elevations of MMP-2 and MMP-9 activity in the hippocampus of rats without or after hypoxia when CPPG was administered. Concluding, CPPG has no effects on defi cits of behavioural processes induced by hypoxia in the water maze. Enhanced activity of MMP-2 and MMP9 in the hippocampus induced by CPPG does not correspond to the infl uence on acquisition in the water maze. The reconfi guration of extracellular matrix by MMP-2 and MMP-9 in hippocampus is new central effect of CPPG. Support: grant No 3-10614L from the State Committee for Scientifi c Research, Warszawa, Poland.
Insulin insufficiency and increased glucose levels are the major features of diabetes type 1 leading to cognitive dysfunctions and neurodegeneration. A reason why different brain structures are characterized by diverse response to increased glucose level is not known. Our previous study showed increased ceramide levels in the brains of rats with diabetes induced by streptozotocin (STZ) injection, which was abolished by myriocin, the inhibitor of serine palmitylotransferase. Ceramides may be important mediators of neuropathological changes and its elevation was found in many brain disorders. The main goal of our present study was to verify if hippocampus, prefrontal cortex and cerebellum response differently to hyperglycemia/hypoinsulinemia in terms of changes in sphingolipids concentrations. We attempted to identify potential source of ceramides by measuring the sphingomyelinase concentrations and by blocking the ceramide de novo synthesis pathway. We found that in cerebellum and hippocampus of hyperglycemic animals sphingolipids concentrations underwent subtle modifications while prefrontal cortex exhibited massive changes in ceramides and SMs content. Total ceramide levels was significantly elevated in prefrontal cortex of diabetic rats, which was reduced by myriocin, while rats exposed to STZ showed only small increase of total SM in this brain structure. The increased content of ceramides containing SAFAs (saturated fatty acids) in prefrontal cortex was diminished by myriocin. SAFA-contained SMs did not present changes. Elevation of MUFA- (monounsaturated fatty acids), and PUFA-ceramides (polyunsaturated fatty acids) in prefrontal cortex of STZ-treated rats was reduced by myriocin, similarly as MUFA-SMs augmentation. PUFA-ceramides and PUFA-SMs experienced only slight modifications. Both – ceramides and omega-6 – SMs increased dramatically and were downregulated by myriocin. We conclude that the prefrontal cortex may be particularly sensitive to hyperglycemic conditions and hypoinsulinemia. Moreover, the novo synthesis seems to be an important pathway of ceramide generation since usage of myriocin strongly reduced ceramide levels enhanced by STZ injection. Augmentation in ceramide content was correlated with enhancement of SMs production. These unexpected results may be explained by the incorporation of redundant ceramides into SMs, a mechanism by which the toxic level of ceramides is reduced in the brain. Supported by grant 123-27575 P from the State Committee for Scientific Research, Warsaw, Poland
Ceramide (Cer) and sphingomyelin (SM) are members of sphingolipid (SL) family. Their concentrationsin the brain undergo substantial changes in many pathologies. They are also important players in diabetes-linked brain dysfunctions, in which increased content of ceramides can be toxic to neurons. The aim of the study was to evaluate selected parameters of sphingolipids and insulin pathways in prefrontal cortex (PC) and hippocampus (H) of rats with experimentally induced hyperglycemia. STZ-rat model of type 1 diabetes and high fat diet model of insulin resistance were used. Analyses of studied parameters were performed by GLC, IHC and Elisa. We found the augmented levels of ceramides in H and PC and only minor in striatum and cerebellum of rats with STZ-induced diabetes. Similar expressions of Cer were confirmed by IHC. Myriocin, an inhibitor of an enzyme of ceramide de novo synthesis pathway, reduced ceramide generation in hyperglycemic brains, particularly in PC, which was reflected in altered Cer synthase activities. In addition, we reported the fluctuations in sphingomyelin levels in investigated structures. The level of insulin did not change in H and PC of STZ-treated rats. An expression of insulin receptor and its phosphorylated form decreased in both structures, but was restored after myriocin administration. Similarly, Akt and phosphorylated Akt changed in these structures suggesting important role of de novo Cer synthesis in intracellular pathways of insulin. In the rat model of high fat diet, which leads to insulin resistance, the sphingolipid pattern (Cer and SM) was altered in H and PC as well. Metformin, the drug of choice in diabetes type 2, influenced the content of the above SLs in these structures, suggesting the additional central activity of antidiabetic treatment. We conclude that ceramide and SM may be important mediators of diabetes- accompanied brain dysfunction.
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