A New and Simple Method for Studying the Microanatomy of Nephrons and Collecting Tubules

A simple procedure for studying the microanatomy of the kidney by light microscopy is described. Specimens are dried by the critical-point method and later impregnated with a mounting medium. The retention of air within the lumen of nephrons and collecting tubules permits their visualization. The method allows the observation of renal microanatomy without the distortions produced by microdissection or the loss of structural relationships among different kidney components. It also permits observation of the same specimen by both light and scanning electron microscopy.     

ECG wave form, short term heart rate variability and plasma catecholamine concentrations in intrauterine growth-retarded guinea-pig fetuses

Electrocardiogram waveform, short term heart rate variability and catecholamine concentrations were studied with maternally-induced anesthesia in eleven growth-retarded guinea-pig fetuses and their normal-sized littermates at 63 days of gestation. Intrauterine growth retardation was induced by unilateral uterine artery ligation performed between day 32 and 35. In the growth-retarded group fetal weight was reduced by 45%. Blood gases, acid-base status and oxygen content were similar in the two groups. The growth-retarded guinea-pig fetuses were hypoglycemic and demonstrated a rise in hemoglobin concentration. The T/QRS ratio (T wave amplitude/QRS amplitude) was similar in both groups. The short-term heart rate variability was significantly reduced in the growth-retarded group. Plasma catecholamine concentrations were increased in growth-retarded fetuses but differed only significantly for noradrenaline compared to controls. We suggest that similar T/QRS ratio in both groups of fetuses indicates that aerobic myocardial metabolism is maintained among growth-retarded fetuses. The mechanism behind the reduced variability is unclear.     

Amyloidogenic Propensity of a Natural Variant of Human Apolipoprotein A-I: Stability and Interaction with Ligands

A number of naturally occurring mutations of human apolipoprotein A-I (apoA-I) have been associated with hereditary amyloidoses. The molecular mechanisms involved in amyloid-associated pathology remain largely unknown. Here we examined the effects of the Arg173Pro point mutation in apoA-I on the structure, stability, and aggregation propensity, as well as on the ability to bind to putative ligands. Our results indicate that the mutation induces a drastic loss of stability, and a lower efficiency to bind to phospholipid vesicles at physiological pH, which could determine the observed higher tendency to aggregate as pro-amyloidogenic complexes. Incubation under acidic conditions does not seem to induce significant desestabilization or aggregation tendency, neither does it contribute to the binding of the mutant to sodium dodecyl sulfate. While the binding to this detergent is higher for the mutant as compared to wt apoA-I, the interaction of the Arg173Pro variant with heparin depends on pH, being lower at pH 5.0 and higher than wt under physiological pH conditions. We suggest that binding to ligands as heparin or other glycosaminoglycans could be key events tuning the fine details of the interaction of apoA-I variants with the micro-environment, and probably eliciting the toxicity of these variants in hereditary amyloidoses.     

More Than a Pore: The Interplay of Pore-Forming Proteins and Lipid Membranes

Pore-forming proteins (PFPs) punch holes in their target cell membrane to alter their permeability. Permeabilization of lipid membranes by PFPs has received special attention to study the basic molecular mechanisms of protein insertion into membranes and the development of biotechnological tools. PFPs act through a general multi-step mechanism that involves (i) membrane partitioning, (ii) insertion into the hydrophobic core of the bilayer, (iii) oligomerization, and (iv) pore formation. Interestingly, PFPs and membranes show a dynamic interplay. As PFPs are usually produced as soluble proteins, they require a large conformational change for membrane insertion. Moreover, membrane structure is modified upon PFPs insertion. In this context, the toroidal pore model has been proposed to describe a pore architecture in which not only protein molecules but also lipids are directly involved in the structure. Here, we discuss how PFPs and lipids cooperate and remodel each other to achieve pore formation, and explore new evidences of protein-lipid pore structures.