Abstract
The purpose of this study was to evaluate the feasibility of a 1H-NMR-based metabolomic approach to explore the metabolomic signature of different topographical areas of vitreous humor (VH) in an animal model. Five ocular globes were enucleated from five goats and immediately frozen at −80°C. Once frozen, three of them were sectioned, and four samples corresponding to four different VH areas were collected: the cortical, core, and basal, which was further divided into a superior and an inferior fraction. An additional two samples were collected that were representative of the whole vitreous body. 1H-NMR spectra were acquired for twenty-three goat vitreous samples with the aim of characterizing the metabolomic signature of this biofluid and identifying whether any site-specific patterns were present. Multivariate statistical analysis (MVA) of the spectral data were carried out, including Principal Component Analysis (PCA), Hierarchical Cluster Analysis (HCA), and Partial Least Squares Discriminant Analysis (PLS-DA). A unique metabolomic signature belonging to each area was observed. The cortical area was characterized by lactate, glutamine, choline, and its derivatives, N-acetyl groups, creatine, and glycerol; the core area was characterized by glucose, acetate, and scyllo-inositol; and the basal area was characterized by branched-chain amino acids (BCAA), betaine, alanine, ascorbate, lysine, and myo-inositol. We propose a speculative approach on the topographic role of these molecules that are mainly responsible for metabolic differences among the as-identified areas. 1H-NMR-based metabolomic analysis has shown to be an important tool for investigating the VH. In particular, this approach was able to assess in the samples here analyzed the presence of different functional areas on the basis of a different metabolite distribution.
Highlights
Over the last years, the vitreous humor (VH) has gained a pivotal role in comprehending and explaining several ocular diseases, and other medical disciplines are even becoming more interested in analyzing this anatomical structure
The aim of the present study was to evaluate the feasibility of a 1H-Nuclear Magnetic Resonance (NMR)-based metabolomic approach to investigate the metabolomic composition of different VH areas in an animal model
The choice of goat samples was driven by the fact that the ocular globes contain approximately 4 ml VH, which is an adequate amount for 1H-NMR analyses of the different topographic areas
Summary
The vitreous humor (VH) has gained a pivotal role in comprehending and explaining several ocular diseases, and other medical disciplines are even becoming more interested in analyzing this anatomical structure. Forensic science scholars are well aware of the importance of its investigation to identify several causes of death and, more importantly, to estimate the time since death (Post-Mortem Interval; PMI) [1] and to diagnose acute and chronic drug intoxication [2] and other causes of death (such as heat-related deaths, hyperglycemia, and dehydration) [3,4]. A proteomic investigation suggested that VH composition may reflect sitespecific features [10]; the same may be hypothesized of a metabolomic signature. To investigate this latter hypothesis, we performed a metabolomic analysis of VH in an animal model
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