Activation of phospholipase hydrolysis during oxidative cell damage

Kondakova, I. V.; Borunov, E. V.; Antipova, E. V.

doi:10.1007/bf00840611

“…In the last few years, our research group has extensively studied the salivary metabolites by liquid and gas chromatography approaches [45][46][47][48][49][50]. The analysis of saliva by ATR-FTIR and Raman provides complementary, fast, and holistic information on the sample, which includes low-molecular-weight (MW) metabolites and (macromolecules proteins, carbohydrates, and lipids), both having a high diagnostic value for local and systemic disorders.…”

Section: Application Preanalytical Stepmentioning

confidence: 99%

“…Fifteen-min at room temperature led to longer oxygen exposure and maybe changed the enzymatic activity in these muscle tissues, which contributed to the oxidation and instability of the unsaturated fatty acids, as well as the generation of their downstream metabolites, i.e., oxylipins [42,43]. The higher lysophospholipids (Table 1), i.e., LPE14.0, LPE16.1, LPE20.4, LPE22.4, LPG16.1, LPI20.4, LPI22.4, and LPI22.6, in 15-min delayed isolation muscle tissues might be due to the hydrolysis of the cellular membrane induced by the longer time of oxidation exposure and oxidative damage [44,45], and/or tissue degeneration. The significantly increased pyruvate in Gas + Sol with delayed isolation (Table 2) may be due to the oxidation of lactate [46].…”

Section: The Effects Of Sample Isolation Speed On Metabolite Stabilitymentioning

confidence: 99%

Advances in Metabolic Profiling of Biological Samples

2023

0

View full text Add to dashboard Cite

show abstract

“…Fifteen-min at room temperature led to longer oxygen exposure and maybe changed the enzymatic activity in these muscle tissues, which contributed to the oxidation and instability of the unsaturated fatty acids, as well as the generation of their downstream metabolites, i.e., oxylipins [42,43]. The higher lysophospholipids (Table 1), i.e., LPE14.0, LPE16.1, LPE20.4, LPE22.4, LPG16.1, LPI20.4, LPI22.4, and LPI22.6, in 15-min delayed isolation muscle tissues might be due to the hydrolysis of the cellular membrane induced by the longer time of oxidation exposure and oxidative damage [44,45], and/or tissue degeneration. The significantly increased pyruvate in Gas + Sol with delayed isolation (Table 2) may be due to the oxidation of lactate [46].…”

Section: The Effects Of Sample Isolation Speed On Metabolite Stabilitymentioning

confidence: 99%

A Sample Preparation Method for the Simultaneous Profiling of Signaling Lipids and Polar Metabolites in Small Quantities of Muscle Tissues from a Mouse Model for Sarcopenia

He

¹

,

Mever

²

,

Yang

³

et al. 2022

View full text Add to dashboard Cite

The metabolic profiling of a wide range of chemical classes relevant to understanding sarcopenia under conditions in which sample availability is limited, e.g., from mouse models, small muscles, or muscle biopsies, is desired. Several existing metabolomics platforms that include diverse classes of signaling lipids, energy metabolites, and amino acids and amines would be informative for suspected biochemical pathways involved in sarcopenia. The sample limitation requires an optimized sample preparation method with minimal losses during isolation and handling and maximal accuracy and reproducibility. Here, two developed sample preparation methods, BuOH-MTBE-Water (BMW) and BuOH-MTBE-More-Water (BMMW), were evaluated and compared with previously reported methods, Bligh-Dyer (BD) and BuOH-MTBE-Citrate (BMC), for their suitability for these classes. The most optimal extraction was found to be the BMMW method, with the highest extraction recovery of 63% for the signaling lipids and 81% for polar metabolites, and an acceptable matrix effect (close to 1.0) for all metabolites of interest. The BMMW method was applied on muscle tissues as small as 5 mg (dry weight) from the well-characterized, prematurely aging, DNA repair-deficient Ercc1∆/− mouse mutant exhibiting multiple–morbidities, including sarcopenia. We successfully detected 109 lipids and 62 polar targeted metabolites. We further investigated whether fast muscle tissue isolation is necessary for mouse sarcopenia studies. A muscle isolation procedure involving 15 min at room temperature revealed a subset of metabolites to be unstable; hence, fast sample isolation is critical, especially for more oxidative muscles. Therefore, BMMW and fast muscle tissue isolation are recommended for future sarcopenia studies. This research provides a sensitive sample preparation method for the simultaneous extraction of non-polar and polar metabolites from limited amounts of muscle tissue, supplies a stable mouse muscle tissue collection method, and methodologically supports future metabolomic mechanistic studies of sarcopenia.

show abstract

“…The manner of acrosome loss revealed by EM was very similar to that reported for treatment with the detergent Triton X-100 (Sistina et al, in press). LPC has been reported to have a detergent-like action on isolated guinea pig ventricular myocytes (Liu et al, 1991) and mastocytoma P815 cells (Kondakova et al, 1991) and was found to lead to disruption of plasma and acrosomal membranes without a n AR in human spermatozoa (Byrd and Wolf, 1986). The acrosomal membrane invagination into the acrosomal matrix that led to vesiculation of the matrix was essentially identical to that seen after calcium ionophore treatment (Mate and Rodger, 19911, freeze-thawing, and detergent treatment (Sistina et al, in press).…”

Section: Ultrastructure Of Lpc-induced Acrosomal Lossmentioning

confidence: 99%

Lysophosphatidylcholine disrupts the acrosome of tammar wallaby (macropus eugenii) spermatozoa

Sistina

¹

,

Lin

²

,

Rodger

³

1993

Molecular Reproduction Devel

View full text Add to dashboard Cite

The acrosomal status of wallaby spermatozoa was evaluated by light and electron microscopy after incubation in 1-100 microM lysophosphatidylcholine (LPC) for up to 120 min. Treatment with 1 and 10 microM LPC for 120 min did not lead to acrosomal loss, or detectable alteration to the acrosome, as detected by Bryan's staining and light microscopy. Incubation with 25 microM LPC had little effect on acrosomal loss, however statistically significant changes (P < 0.05) in the acrosomal matrix (altered) were detected after 10-min incubation by light microscopy. Around 50% of acrosomes were altered after 20-min incubation in 50 microM LPC (P < 0.001), and 40% of spermatozoa had lost their acrosome after 60-min incubation (P < 0.001). Treatment with 75 and 100 microM LPC led to rapid acrosomal loss from around 50% of spermatozoa within 10 min (P < 0.001), and by 60 min acrosomal loss was 70-80%. LPC, like the diacylglycerol DiC8 (1,2-dioctanoyl-sn-glycerol), is thus an effective agent to induce loss of the relatively stable wallaby sperm acrosome, and it also induces changes within the acrosomal matrix. Ultrastructure of the LPC-treated spermatozoa revealed that the plasma membrane and the acrosomal membranes were disrupted in a manner similar to that seen after detergent treatment (Triton X-100). There was no evidence of point fusion between the plasma membrane overlying the acrosome and the outer acrosomal membrane. The plasma membrane was the first structure to disappear from the spermatozoa. The acrosomal membranes and matrix showed increasing disruption with time and LPC concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

Activation of phospholipase hydrolysis during oxidative cell damage

Cited by 4 publications

References 11 publications

Advances in Metabolic Profiling of Biological Samples

Advances in Metabolic Profiling of Biological Samples

A Sample Preparation Method for the Simultaneous Profiling of Signaling Lipids and Polar Metabolites in Small Quantities of Muscle Tissues from a Mouse Model for Sarcopenia

Lysophosphatidylcholine disrupts the acrosome of tammar wallaby (macropus eugenii) spermatozoa

Contact Info

Product

Resources

About