Unraveling the association between genetic integrity and metabolic activity in pre-implantation stage embryos

D’Souza, Fiona; Pudakalakatti, Shivanand; Uppangala, Shubhashree; Honguntikar, Sachin D.; Salian, Sujith Raj; Kalthur, Guruprasad; Pasricha, Renu; Appajigowda, Divya; Atreya, Hanudatta S.; Adiga, Satish Kumar

doi:10.1038/srep37291

Cited by 18 publications

(20 citation statements)

References 44 publications

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“…The induction of CDDP mediated DNA lesions at the 2-cell stage affected blastocyst formation and hatching potential. These results are in agreement with our previous observation, where embryos carrying induced DNA lesions (IDL) developed normally for the first 2.5 days (60 hpi-hCG), but began to exhibit a developmental delay at 84 hpi-hCG, though they were morphologically indistinguishable from control embryos 8 . On the other hand, the potential of hatched blastocysts to attach on Ex 24 and develop into ICM outgrowths was similar in both control and IDL groups.…”

Section: Discussionsupporting

confidence: 93%

“…Based on previous results 8 , cis-Diamineplatinum(II) dichloride (Cisplatin, CDDP) was used to induce DNA lesions in 2-cell stage mouse embryos. At the 2-cell stage, embryos were randomly assorted to control (N = 392) and exposed to 3 µM CDDP which constituted the IDL group (N = 675).…”

Section: Resultsmentioning

confidence: 99%

“…Early detection of changes in metabolic pattern has the potential to provide an immense advantage in aiding the selection of a competent embryo with high implantation potential 8 , 23 – 25 . Intensities of metabolites in the spent medium was interpreted as uptake or release, compared to the level of metabolites in culture medium without embryos which was used to determine the baseline of metabolite levels 26 .…”

Section: Resultsmentioning

confidence: 99%

“…Preimplantation stage embryos are sensitive to exogenously induced DNA lesions such as radiation 6 and chemotherapeutic agents 7 , 8 and this could be due to a peculiarity in the damage responses of the early-stage embryos 9 – 11 . Due to species specific and embryonic stage specific differences, cell cycle regulations vary between somatic and embryonic cells within a species.…”

Section: Introductionmentioning

confidence: 99%

“…In a similar retrospective approach, we were earlier able to demonstrate that biomarkers from 84 hours post injection of hCG (hpi-hCG) spent embryo culture medium could be used to predict the genetic integrity and blastocyst development 24 h in advance, using Nuclear Magnetic Resonance (NMR) spectroscopy 8 . However, these observations were limited to the developmental competence and genetic integrity of embryos until the blastocyst stage.…”

Section: Introductionmentioning

confidence: 99%

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Spent embryo culture medium metabolites are related to the in vitro attachment ability of blastocysts

D’Souza

Uppangala

Asampille

et al. 2018

Sci Rep

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The metabolomic profile of an embryo culture medium can aid in the advanced prediction of embryonic developmental potential and genetic integrity. But it is not known if this technology can be used to determine the in vitro potential of inner cell mass (ICM) in adherence and proliferation. Here, we investigated the developmental potential of mouse 2-cell embryos carrying cisplatin-induced DNA lesions (IDL), beyond blastocyst stage using ICM outgrowth assay. The genetic integrity of ICM cells was determined by comet assay. The metabolic signatures of spent medium were recorded 84 hours post injection of hCG (hpi-hCG), and after 96 hours of extended in vitro culture (Ex 96) by NMR spectroscopy. We observed that blastocysts that lack the ability to adhere in vitro had an increased requirement of pyruvate (p < 0.01), lactate (p < 0.01), and were accompanied by a significant reduction of pyruvate-alanine ratio in the culture medium. We propose that the aforementioned metabolites from 84 hpi-hCG spent medium be further explored using appropriate experimental models, to prove their potential as biomarkers in the prediction of implantation ability of in vitro derived human embryos in clinical settings.

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Section: Discussionsupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Spent embryo culture medium metabolites are related to the in vitro attachment ability of blastocysts

D’Souza

Uppangala

Asampille

et al. 2018

Sci Rep

Self Cite

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Unlocking the Chemical Diversity of Plant Catharanthus roseus: Nuclear Magnetic Resonance Spectroscopy Approach

Gupta,

Kavya,

Jayalakshmi

et al. 2024

Magnetic Reson in Chemistry

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Catharanthus roseus, also known as Madagascar periwinkle, is a perennial plant renowned for its extensive pharmacological properties. It produces vital chemotherapeutic compounds, including vinblastine and vincristine, and exhibits anti‐inflammatory, antidiabetic, and antioxidant activities. In this study, we utilized a range of two‐dimensional (2D) nuclear magnetic resonance (NMR) techniques, such as 1H–1H correlation spectroscopy (COSY), 1H–1H J‐resolved NMR, and 1H–13C heteronuclear single quantum coherence (HSQC) sensitivity‐enhanced NMR spectroscopy, to identify key metabolites in C. roseus leaf extracts. Given the presence of numerous metabolites with closely spaced multiplet resonances, the 1H NMR spectra often exhibit significant signal overlap, making metabolite identification difficult or even impossible. However, the use of 2D NMR techniques effectively overcame this challenge, allowing for the precise identification of important alkaloids, such as vindoline, vinblastine, serpentine, and ajmalicine, along with essential metabolites like organic acids, amino acids, and carbohydrates. The extract contained a variety of bioactive compounds, including organic acids crucial for the tricarboxylic acid (TCA) cycle, branched‐chain amino acids vital for metabolic functions, and alkaloids with substantial therapeutic potential. This comprehensive study underscores the continued significance of C. roseus in both traditional and modern medicine, emphasizing its intricate metabolic network and its potential in the development of novel therapeutics.

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SOFAST-HMQC—an efficient tool for metabolomics

2017

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Nuclear magnetic resonance (NMR)-based metabolomics relies mostly on 1D NMR; however, the technique is limited by overlap of the signals from the metabolites. In order to circumvent this problem, 2D H-C correlation spectroscopy techniques are often used. However owing to poorer natural abundance and gyromagnetic ratio of C, the acquisition time for 2DH-C heteronuclear single quantum coherence spectroscopy (HSQC) is long. This makes it almost impossible to be used in high throughput study. We have reported the application of selective optimized flip angle short transient (SOFAST) technique coupled to heteronuclear multiple quantum correlation (HMQC) along with nonlinear sampling (NUS) in urine and serum samples. This technique takes sevenfold less experimental time than the conventional H-C HSQC experiment with retention of almost all molecular information. Hence, this can be used for high throughput study. Graphical abstract SOFAST-HMQC is a two-dimensional NMR technique that significantly decreases experimental time without loss of information. This technique is applied in complex biofluid samples that are used for high throughput metabolomics studies and shows promise of better information recovery than conventional two-dimensional NMR technique in shorter time.

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Unraveling the association between genetic integrity and metabolic activity in pre-implantation stage embryos

Cited by 18 publications

References 44 publications

Spent embryo culture medium metabolites are related to the in vitro attachment ability of blastocysts

Spent embryo culture medium metabolites are related to the in vitro attachment ability of blastocysts

Unlocking the Chemical Diversity of Plant Catharanthus roseus: Nuclear Magnetic Resonance Spectroscopy Approach

SOFAST-HMQC—an efficient tool for metabolomics

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