2‐D DIGE and MS/MS analysis of protein serum expression in rats housed in concrete and clay cages in winter

Abstract

In a previous study, we examined the physiological responses of male Sprague-Dawley rats over a 4-week exposure to concrete and clay cages. No general toxicological changes were observed in rats exposed to either of the two cage types in summer. Under winter conditions, however, various general toxicological effects were detected in rats housed in concrete cages, although rats housed in clay cages showed no such effects. The infrared thermographic examination indicated that skin temperature in the concrete-housed rats was abnormally low, but not so in the clay-housed rats. We examined proteomic changes in the serum of rats housed in winter in concrete and clay cages using two-dimensional differential in-gel electrophoresis and mass spectrometry/mass spectrometry. Five proteins were identified and quantitatively validated; all were cold stress-induced, acute phase proteins that were either up-regulated (haptoglobin) or down-regulated (alpha-1-inhibitor III, alpha-2u globulin, complement component 3, and vitamin D-binding protein) in the concrete-housed rats. These results suggest that the 4-week exposure to a concrete cage in winter elicited a typical systemic inflammatory reaction (i.e. acute phase response) in the exposed rats.

Mass spectrometry of peptides and proteins from human blood

“…The use of propyl sulfate chromatography of intact proteins followed by high resolution, 2D IEF/SDS page gels produced only approximately 300 proteins by MASCOT (Pieper et al, 2003a). Potential biomarkers have been detected using 2D-PAGE methods and in some cases confirmed by independent immunological methods (Dowling et al, 2007;Jackson et al, 2007;White et al, 2007;Barba de la Rosa et al, 2008;Jmeian & El Rassi, 2008;Kim et al, 2008a;Kolialexi et al, 2008;Schlautman et al, 2008;Atkinson et al, 2009;Hamrita et al, 2009). Shifts in observed mass or iso-electric point may reflect physiologically significant processing and or posttranslational modifications (Kapinsky et al, 2001;Johnson et al, 2007).…”

“…Analysis of plasma from patients with squamous cell carcinoma of the uterus revealed potential biomarkers confirmed by Western blot ). Thus, comparison of chromatographically prepared blood fluids by 2D electrophoresis has yielded many changes in the amounts, modifications or processing of relatively abundant blood proteins (Barba de la Rosa et al, 2008;Atkinson et al, 2009), revealing the presence of cancerous or pre cancerous states (He et al, 2003, preeclampsia , or the response to environmental factors (Kim et al, 2008a). The use of novel electrophoresis buffers and mediums may permit the analysis of treatments and protein interactions in blood fluids Manabe, Jin, & Tani, 2007).…”

Proteomic analysis reveals oxidative stress response as the main adaptative physiological mechanism in cows under different production systems

“…In this regard, several aspects of cattle management have also been shown to alter redox markers, such as diet [24,25], transport [26] and in transition dairy cows [27]. It has been also described that cold stress induces an acute phase reaction in rats (and indeed cows living in the mountains were more exposed to cold than cows living close to humans) [28]. The increased levels of fecal corticosterone in Bruna cows living in the mountains compared to those living in better conditions indicates that these animals indeed suffer a certain degree of chronic stress.…”