Seasonal changes and steroid control of perineuronal nets in the song control system

Perineuronal nets (PNNs) are aggregations of extracellular matrix components surrounding the soma of some neurons, mainly GABAergic interneurons expressing the calcium binding protein parvalbumin (PV+). In mammals, the development of PNNs limits synaptogenesis around PV+ neurons and PNNs have been validated as a marker characterizing the end of critical periods for some types of learning. In oscines, song learning is limited to critical periods either during ontogeny in close-ended learners such as zebra finches or during specific phases of the annual cycle in open-ended learners such as canaries or starlings. In zebra finches, PNN expression increases when the song crystalizes and this increase is markedly inhibited if juveniles are deprived from a tutor, which is known to delay the closure of the critical period for song learning. Nothing is known however about a possible role of PNNs in adult seasonal plasticity of open-ended learners. We compared PNNs expression and their colocalization with PV+ neurons in photosensitive, photostimulated, and photorefractory starlings. Although this treatment affected as expected the testes volumes, testosterone concentrations and volumes of song control nuclei, it did not markedly change the expression of PNNs or PV+ neurons in song control or auditory nuclei. In a second experiment, brains of female canaries implanted with testosterone for 1, 2, 9, or 21 days displayed an increase of the total numbers of PV+ neurons and PNNs in HVC, the total number of PNNs in nucleus robustusarcopallialis (RA), the density of PNNs in Area X and the %PV+ neurons surrounded by PNNs in RA and Area X. Interestingly the density of PNNs in song control nuclei progressively decreases from zebra finches to canaries to starlings in parallel with the increased song plasticity in these species supporting the notion that PNNs may limit brain and thus song plasticity in a species-typical manner.