Civilization diseases associated with memory disorders are important health problems occurring due to a prolonged life span. The manuscript shows the results of an in vivo study targeting the emergence of two drug candidates with anti-amnestic properties. The preceding quantitative structure–activity relationship (QSAR) studies provided information on the ability of berberine and magnoflorine to cross the blood–brain barrier (BBB). In the light of these findings, both compounds were purified from crude plant extracts of barberries: berberine—from Berberis siberica using a method published earlier, and magnoflorine—from Berberis cretica by centrifugal partition chromatography (solvent system: ethyl acetate:butanol:water-0.6:1.5:3 v/v/v). Both the compounds were evaluated for their memory enhancing and scopolamine inhibitory properties in an in vivo passive avoidance (PA) test on mice towards short-term and long-term memory. Cognition enhancing properties were observed at the following doses: 5 mg/kg (i.p.) for berberine and 20 mg/kg (i.p.) for magnoflorine. In addition, both the tested isoquinolines with the co-administered scopolamine were found to block long-term but not short-term memory impairment. No influence on the locomotor activity was observed for the tested doses. The results confirmed a marked central activity of magnoflorine and showed the necessity to lower the dosage of berberine. Optimized purification conditions have been elaborated for magnoflorine.
Calretinin (CR) as a buffer and sensor protein plays an important role in regulatory processes of Ca 2+ and anty-apoptotic cellular protection. In the present study, immunohistochemical peroxidase-antiperoxidase (PAP) method was used in order to determine the numbers, morphology, morphometry and distribution pattern of CR in neurons of the chinchilla's claustrum (Cl) and endopiriform nucleus (EN). In Cl and EN the presence of several classes of neurons with different immunoreactivity to CR was found. In Cl, CR-immunoreactive (IR) neurons were predominantly found in close vicinity to insular border while CR-IR neurons were evenly scattered throughout EN. In general, immunoreaction to CR was observed in neuronal cytoplasm, nucleus and in fibres-like nerve extensions. Statistical analysis showed the differences between average large diameter as well as cross-sectional area of CR-IR neurons present in Cl and EN. It is suggested, that CR acting as a calcium binding protein may play a role in neuronal network. Further co-localization studies are necessary to fully elucidate neurophysiology and neuropathology of the chinchilla's Cl and EN neurons.
Influence of oral administration of HMB to pregnant dams on calbindin expression in the dentate gyrus of the hippocampus during postnatal development in spiny mice offspring Summary The aim of the study was to investigate the morphology, density and immunostaining intensity of calbindin (CB)-positive neurons of dentate gyrus (DG) in new-born (P0) and 21-day-old (P21) male Acomys cahirinus mice from dams receiving β-hydroxy-β-methylbutyrate (HMB) during pregnancy. Different substances administrated to pregnant dams may affect the calcium ion homeostasis which is crucial for the proper brain development of their offspring. DG with hilus (H) plays an important role in memory and learning processes. Calcium levels in DG are regulated by buffering proteins like calbindin D28k (CB). Experimental dams were orally treated with HMB at a dose of 0.2 g/kg b.w. Half of new-born animals were euthanised after birth and the rest after the 21 st day of life. The brains were dissected and embedded in paraffin blocks using a routine histological technique. In order to demonstrate CB protein expression an immunohistochemical peroxidase-antiperoxidase reaction was conducted. The results of the study did not reveal important morphological alterations. There were no statistically significant changes in the density of the studied cells either in P0 and P21 animals. However, the authors have demonstrated a statistically significant increase of the average CB-immunostaining intensity in nuclei and cytoplasm in both age groups. It may be a result of a compensation effect to alterations that occurred under the influence of HMB. On the basis of the conducted research, it may be assumed that HMB activity in DG may provide long-term consequences.
AbstractαCaMKII, widely occurring in the central nervous system, plays a significant role in cognitive processes. It is well known that diabetes is a risk factor that may trigger brain atrophy, cognitive dysfunction and finally lead to memory loss. Antioxidants richly present in bilberry fruits are believed to have significant effects on diabetes-related brain dysfunctions mainly due to their abilities to modulate neurotransmitter release that lead to reduction of the negative impact of free radicals on cognitive processes. The aim of the present research was to immunohistochemically investigate the expression patterns of αCaMKII in hippocampal neurons from non-diabetic, diabetic and diabetic rats fed with an extract of bilberry fruit. The obtained results show that in comparison to the control group, in diabetic rats hippocampal neurons immunoreactive (ir) to αCaMKII were swollen and the lengths of the neuronal fibres were reduced. Further study shows that in diabetic rats fed with bilberry fruit, αCaMKII-positive nerve fibres were significantly longer when compared to the groups of diabetic and control rats. Additionally, we observed statistically significant changes in the average larger diameter of αCaMKII-ir hippocampal neurons between groups of diabetic rats (with vs. without supplement of bilberry fruit). The results of the present work suggest that antioxidants present in bilberry fruits influence the morphology of and possibly exhibit beneficial and neuroprotective effects on hippocampal neurons during diabetes. It is likely that changes in the appearance of αCaM-KII-expressed hippocampal neurons may reflect the diabetes-evoked rise in Ca 2+ level in the cerebral nerve terminals. The present research extends our knowledge of preventive mechanisms for cognitive dysfunctions occurring in the brain during diabetes.
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