Early-life metabolic dysfunction impairs cognition and mitochondrial function in mice.

The impact of overnutrition early in life is not restricted to the onset of cardiovascular and metabolic diseases, but also affects critical brain functions related to cognition. This study aimed to evaluate the relationship between peripheral metabolic and bioenergetic changes induced by a two-hit protocol and their impact on cognitive function in juvenile mice. Three-week-old male C57BL/6 mice received a high-fat diet (HFD) or control diet for 7 weeks, associated with two low doses of streptozotocin (STZ) or vehicle. Despite the absence of obesity, HFD+STZ impaired glucose metabolism and induced a trend towards cholesterol increase. The two-hit protocol impaired recognition and spatial memories in juvenile mice, without inducing a depressive-like behavior. HFD+STZ mice presented increased immunoreactivity for GFAP and a trend towards a decrease in NeuN in the hippocampus. The treatment caused a bioenergetic impairment in the hippocampus, characterized by a decrease in both O2 consumption related to ATP production and in the maximum respiratory capacity. The thermogenic capacity of brown adipose tissue was impaired by the two-hit protocol, here verified through the absence of a decrease in O2 consumption after uncoupled protein-1 inhibition and an increase in the reserve respiratory capacity. Impaired mitochondrial function was also observed in the liver of HFD+STZ juvenile mice, but not in their heart. These results indicate that exposure to HFD+STZ early in life has a detrimental impact on the bioenergetic and mitochondrial function of tissues with metabolic and thermogenic activities, which is likely related to hippocampal metabolic changes and cognitive impairment.

Application based on the Canny edge detection algorithm for recording contractions of isolated cardiac myocytes.

BACKGROUND AND OBJECTIVE: The isolated cardiomyocyte preparation is amenable to several experimental approaches not suitable to the myocardial tissue, which has allowed the gain of important information on the pathophysiology of the cardiac muscle. Thus, the development of techniques for functional studies in this preparation is important. The goal of the present study was to develop a computer program to extract contraction traces generated by cyclic cell shortening from cardiomyocyte video image files. METHODS: The Canny algorithm, widely used for computer vision, was implemented for cell edge recognition and continuous tracking, so that changes in cardiomyocyte length could be monitored. The program was applied to demonstrate the effect of classical inotropic maneuvers on contraction parameters, as well as to assess the development of spontaneous activity in response to defibrillator-like electrical shocks in rat isolated cardiomyocytes. RESULTS: The method resulted in successful monitoring of variations in cell length during both electrically-triggered and post-shock spontaneous contractions, of which the rate was significantly related to shock strength. CONCLUSIONS: The proposed approach might be useful for analysis of contractile activity of isolated muscle cells, and allows detection of even the typically low-amplitude noisy spontaneous contractile events. Additionally, the experimental data suggest that the rate of spontaneous contraction could be used as an index of shock-induced electrical membrane damage.

The influence of cell dimensions on the vulnerability of ventricular myocytes to lethal injury by high-intensity electrical fields / Influência das dimensões celulares sobre a vulnerabilidade de miócitos ventriculares ao efeito letal de campos elétricos de alta intensidade

Application of high intensity electric fields (HIEF) to the myocardium is commonly used for cardiac defibrillation/cardioversion. Although effective at reversing life-threatening arrhythmias, HIEF may cause myocyte damage due to membrane electropermeabilization. In this study, the influence of cell length and width on HIEF-induced lethal injury was analyzed in isolated rat cardiomyocytes in parallel alignment with the field. The field-induced maximum variation of membrane potential (ΔVmax) was estimated with the Klee-Plonsey model. The studied myocyte population was arranged in two group pairs for comparison: the longest vs. the shortest cells, and the widest vs. narrowest cells. Threshold field intensity was significantly lower in the longest vs. shortest myocytes, whereas cell width influence was not significant. The threshold ΔVmax was comparable in all groups. Likewise, a significant leftward shift of the lethality curve (i.e., relationship of the probability of lethality vs. field intensity) of the longest cells was observed, evidencing greater sensitivity to HIEF-induced damage. However, the lethality curve as a function of ΔVmax was similar in all groups, confirming a prediction of the Klee-Plonsey model. The similar results for excitation and injury at threshold and HIEF stimulation, respectively, indicate that: a) the effect of cell length on the sensitivity to the field would be attributable to differences in field-induced membrane polarization that lead to excitation or lethal electroporation; b) the Klee-Plonsey model seems to be reliable for analysis of cell interaction with HIEF; c) it is possible that increased cell length in hypertrophied hearts enhances myocyte fragility upon defibrillation/cardioversion.

Campos elétricos de alta intensidade (HIEF) são aplicados ao miocárdio durante desfibrilação e cardioversão. Embora eficazes na reversão de arritmias potencialmente letais, HIEF podem lesar cardiomiócitos por eletropermeabilização da membrana. Neste estudo, a influência das dimensões celulares sobre o efeito letal de HIEF foi estudada em cardiomiócitos isolados de rato alinhados paralelamente ao campo. A máxima variação do potencial de membrana induzida pelo campo (ΔVmax) foi calculada com o modelo de Klee-Plonsey. As células estudadas foram distribuídas em dois pares de grupos de acordo com seu comprimento e largura. A intensidade limiar do campo não dependeu da largura celular, mas sim do comprimento (menor nas células mais longas, p < 0.001), enquanto ΔVmax no limiar foi comparável entre os grupos. Nas células mais longas, observou-se desvio à esquerda (p < 0.01) da curva que descreve a relação entre probabilidade de letalidade e a intensidade do campo, evidenciando maior sensibilidade à ação deletéria de HIEF. Porém, a curva de letalidade em função de ΔVmax foi semelhante em todos os grupos, o que confirma a predição pelo modelo de Klee-Plonsey. A similaridade de resultados com estimulação limiar e com HIEF indica que: a) o efeito do comprimento celular sobre a sensibilidade ao campo poderia ser atribuído a diferenças no grau de polarização da membrana durante a aplicação do estímulo; b) o modelo de Klee-Plonsey parece ser confiável para a análise da interação espacial da célula com HIEF; c) é possível que o maior comprimento celular em miócitos hipertrofiados os torne mais susceptíveis a lesão durante desfibrilação/cardioversão.