Using in vivo calcium imaging to examine joint neuron spontaneous activity and home cage analysis to monitor activity changes in mouse models of arthritis

Abstract

Studying pain in rodent models of arthritis is challenging. For example, assessing functional changes in joint neurons is challenging due to their relative scarcity amongst all sensory neurons. Additionally, studying pain behaviors in rodent models of arthritis poses its own set of difficulties. Commonly used tests, such as static weight-bearing, often require restraint, which can induce stress and consequently alter nociception. The aim of this study was to evaluate two emerging techniques for investigating joint pain in mouse models of rheumatoid- and osteo-arthritis: In vivo calcium imaging to monitor joint afferent activity and group-housed home cage monitoring to assess pain-like behaviors. Specifically, we examined whether there was increased spontaneous activity in joint afferents and reduced locomotor activity following induction of arthritis. Antigen induced arthritis (AIA) was used to model rheumatoid arthritis and partial medial meniscectomy (PMX) was used to model osteoarthritis. Group-housed home cage monitoring was used to assess locomotor behavior in all mice, and weight bearing was assessed in PMX mice. In vivo calcium imaging with GCaMP6s was used to monitor spontaneous activity in L4 ganglion joint neurons retrogradely labelled with fast blue 2 days following AIA and 13–15 weeks following PMX model induction. Cartilage degradation was assessed in knee joint sections stained with Safranin O and fast green in PMX mice. Antigen induced arthritis produced knee joint swelling and PMX caused degeneration of articular cartilage in the knee. In the first 46 h following AIA, mice travelled less distance and were less mobile compared to their control cage mates. In contrast, no such differences were found between PMX and sham mice when measured between 4–12 weeks post-surgery. A larger fraction of joint neurons showed spontaneous activity in AIA but not PMX mice. Spontaneous activity was mostly displayed by medium-sized neurons in AIA mice and was not correlated with any of the home cage behaviors. Group-housed home cage monitoring revealed locomotor changes in AIA mice, but not PMX mice (with n = 10/group). In vivo calcium imaging can be used to assess activity in multiple retrogradely labelled joint afferents and revealed increased spontaneous activity in AIA but not PMX mice.