ADD10 protects renal cells from cold injuries by improving energy metabolism

Cassim, Shamir; Martin, Pierre‐Yves; Pascolo-Rebouillat, Emanuelle

doi:10.1016/j.bbrc.2022.10.009

Cited by 1 publication

(3 citation statements)

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“…2A, C). It was previously shown that basal and maximal respiration rates are proportional to cell viability (Armstrong et al, 2018;Cassim et al, 2022). Apparently, small cell viability differences do not significantly affect respiration parameters.…”

Section: Resultsmentioning

confidence: 89%

“…Our study revealed a notable association between slight differences in cell viability, as determined by the trypan blue exclusion test, and significant effects on uncoupled respiration stability. Basal respiration of trypan-positive cells with either digitonin-permeabilized (Horbay et al, 2012;Pesta & Gnaiger, 2012;Rose et al, 2019) or otherwise damaged (Cassim et al, 2022) plasma membrane is usually more than 5-fold lower compared to intact cells due to metabolite efflux from cytoplasm. The respiration of permeabilized pancreatic acinar cells can be restored by the addition of exogenous oxidative substrates and ADP .…”

Section: Resultsmentioning

confidence: 99%

“…Mitochondrial functional activity in cells is highly correlated with plasma membrane integrity. In renal cells, cold injury causes disruption of cell morphology, a 70% lactate dehydrogenase activity release, and nearly complete loss of both basal and uncoupled cell respiration (Cassim et al, 2022). In cardiac myocytes, hypoxia-reoxygenation induces a reduction of maximal uncoupled respiration in parallel with an increase of cell necrosis measured with a combination of calcein-AM and plasma membrane impermeant ethidium homodimer-1 (Pfleger et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Uncoupled respiration stability of isolated pancreatic acini as a novel functional test for cell vitality

Zub,

Manko,

Manko

et al. 2023

Biol. Stud.

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Background. Assessment of cell viability is crucial in cell studies. Testing plasma membrane integrity is a traditional approach of evaluating cell viability. Mitochondrial functional capacity closely correlates with plasma membrane integrity and overall cell health. This study aimed to investigate whether any aspect of mitochondrial adaptive capacity in isolated pancreatic acini is associated with the quality of isolated pancreatic acini preparations, as determined by the dye exclusion method. Materials and Methods. Experiments were carried out on male Wistar rats weighing 250–300 g. A suspension of isolated pancreatic acini was obtained using collagenase. The rate of oxygen consumption of rat isolated pancreatic acini was measured with Clark oxygen electrode. Basal respiration of isolated pancreatic acini was recorded for approximately 2 min. Afterwards, the mitochondrial adaptive capacity was examined using FCCP in concentrations from 0.5 to 2 μM. Uncoupled respiratory stability was calculated as a ratio of respiration rate at high and low FCCP concentrations. Plasma membrane integrity was assessed with trypan blue staining. A total of 74 preparations of isolated pancreatic acini were used in this study. Results. In all experiments, 92–99 % of pancreatic acinar cells exhibited negative trypan blue staining, indicating intact plasma membranes. The basal and maximal uncoupled respiration rates were not affected by the fraction of trypan-negative cells. However, acini preparations with <less than 95 % plasma membrane integrity had significantly lower uncoupled respiration rates when exposed to a high concentration of FCCP (2 µM), indicating reduced stability of uncoupled respiration. Conclusions. Results of the study suggest that the stability of uncoupled respiration can serve as a novel metabolic functional test to complement the existing methods for assessing cell vitality.

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Section: Resultsmentioning

confidence: 89%