Abstract
Astyanax mexicanus consists of eyed, river-dwelling "surface" fish, and multiple, independently evolved cave populations, which have converged on troglobitic traits such as eye loss and reduced metabolism. However, considerably less is known about constructive adaptations, which include a larger olfactory epithelium in cavefish. It is unknown how this relates to the olfactory bulb (OB), which is the first stage of olfactory sensory processing in the brain. The goal of the present study is to begin to define the structure and functional organization of the OB in A. mexicanus, and to begin to understand how it was transformed via cave adaptation. We addressed these questions using whole-mount immunohistochemistry and in vivo Ca(2+) imaging from the OB of developmentally matched surface and Pachón cavefish. The cavefish OB was significantly larger than surface fish by 14 days post fertilization (dpf), which was accompanied by a broad and proportional increase in synaptic input to most glomerular regions. Increases in the size of the OB were accompanied by increases in the number of neurons expressing tyrosine hydroxylase and calretinin, the latter of which occurred primarily in the medial OB and could not be explained as a compensatory response to a larger OB. In vivo Ca(2+) imaging from the dorsal OB of surface and cavefish in response to a panel of chemical stimuli revealed odor-evoked responses that were spatially organized and highly conserved across the two populations. Surprisingly, the medial OB was consistently activated by any change in water flow in both populations, although the number of water-responsive neurons was significantly greater in cavefish when measurements were performed using either in vivo imaging or the neuronal activity marker phospho-ERK. Water-responding neurons were similarly present in the olfactory epithelium in both populations, along with neurons expressing the mechanosensitive ion channel Piezo2, with significantly more Piezo2-expressing neurons present in cavefish. Therefore, cavefish exhibit enhanced multisensory integration of olfactory and mechanosensory input in the earliest stage of olfactory sensory processing in the brain.