Proteomic profiling of proteins in the dorsal horn of the spinal cord in dairy cows with chronic lameness

Abstract

Chronic lameness affects bovine welfare and has a negative economic impact in dairy industry. Moreover, due to the translational gap between traditional pain models and new drugs development for treating chronic pain states, naturally occurring painful diseases could be a potential translational tool for chronic pain research. We therefore employed liquid chromatography tandem mass spectrometry (LC-MS/MS) to stablish the proteomic profile of the spinal cord samples from lumbar segments (L2-L4) of chronic lame dairy cows. Data were validated and quantified through software tool (Scaffold® v 4.0) using output data from two search engines (SEQUEST® and X-Tandem®). Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis was performed to detect proteins interactions. LC-MS/MS identified a total amount of 177 proteins; of which 129 proteins were able to be quantified. Lame cows showed a strong upregulation of interacting proteins with chaperone and stress functions such as Hsp70 (p < 0.006), Hsc70 (p < 0.0079), Hsp90 (p < 0.015), STIP (p > 0.0018) and Grp78 (p <0.0068), and interacting proteins associated to glycolytic pathway such as; γ-enolase (p < 0.0095), α-enolase (p < 0.013) and hexokinase-1 (p < 0.028). It was not possible to establish a clear network of interaction in several upregulated proteins in lame cows. Non-interacting proteins were mainly associated to redox process and cytoskeletal organization. The most relevant down regulated protein in lame cows was myelin basic protein (MBP) (p < 0.02). Chronic inflammatory lameness in cows is associated to increased expression of stress proteins with chaperone, metabolism, redox and structural functions. A state of endoplasmic reticulum stress and unfolded protein response (UPR) might explain the changes in protein expression in lame cows; however, further studies need to be performed in order to confirm these findings.

Changes in serum biomarkers of inflammation in bovine besnoitiosis

“…In addition, higher antibody levels have been associated with severely chronically affected animals [ 37 ]. Moreover, high values of globulins have been described in cows with chronic diseases [ 38 ], high values of total proteins in cows diagnosed with endometritis [ 39 ] and chronic lameness [ 40 ]. Globulin concentrations above 50 g/l are an indicator of chronic inflammation [ 41 ].…”

The Neuroimmune Interface and Chronic Pain Through the Lens of Production Animals

“…Neuroimmune contributions to chronic pain associated with lameness have recently been confirmed in sheep, goats ( Deng et al, 2018 ), and cattle ( Herzberg et al, 2020a , b ). The lumbar spinal cord dorsal horn in chronically lame dairy cows had significantly higher concentrations of several cytokines (IL-1α, IL-13, IFN-γ, IFN-α, CXCL10, CXCL9, and TNF-α) compared to non-lame counterparts ( Herzberg et al, 2020b ).…”

“…There has been increasing interest in examining chronic pain and the neuroimmune consequences of injury and disease in livestock production and welfare. While still in its infancy there have been a few studies investigating the cellular, molecular, and genetic influences on the development of pain following husbandry procedures ( Sandercock et al, 2019 ) and disease ( Deng et al, 2018 ; Herzberg et al, 2020a ). The fundamental principles of the neuroimmune interface that we are hypothesizing build upon the wealth of knowledge that is currently available to neuroimmune hypothesis in rodents and in humans and will likely have relevance to the livestock sector.…”

“…In production animals, such as sheep, cattle, and pigs, the term chronic pain is used more broadly, often to describe pain persisting past the point of tissue healing or past the acute phase ( Kent et al, 1999 ; McCracken et al, 2010 ; McLennan, 2018 ; Williams, 2019 ; Herzberg et al, 2020a ; Adcock, 2021 ), with less focus on a specified time point. A well-recognized example of chronic pain in livestock is lameness caused by various pathogens and environmental factors including Dichelobacter nodosus causing footrot in sheep and osteoarthritis of infectious, traumatic, or degenerative origin ( Herzberg et al, 2020a ). A more unique aspect of the life of a farm animal compared to humans, is the necessary husbandry practices required to manage animals in a production system.…”

“…Research into the long-lasting effects of painful husbandry procedures in livestock is limited ( Viñuela-Fernández et al, 2007 ; Herzberg et al, 2020b ; Small et al, 2021 ). However, growing interest in improving the welfare of animals is driving further investigation of the long-term consequences of various husbandry practices and chronic disease in livestock ( Troncoso et al, 2018 ; Larrondo et al, 2019 ; Sandercock et al, 2019 ; Herzberg et al, 2020a ).…”