Steroid sex hormones induce dramatic seasonal changes in reproductive related behaviors and their underlying neural substrates in seasonally breeding vertebrates. For example, in adult white-crowned sparrows, increased Spring photoperiod raises circulating testosterone, causing morphological and electrophysiological changes in song-control nuclei, which modify song behavior for the breeding season. We investigated how photoperiod and steroid hormones induce these changes in morphology, electrophysiology, and behavior. Neurons in a song premotor nucleus, the robust nucleus of the arcopallium (RA), show increased intrinsic spontaneous firing rate and soma size when birds are in breeding condition. Using combinations of systemic and unilateral local intracerebral hormonal manipulations, we show that long-day photoperiod accelerates the effects of systemic testosterone on RA neurons via the estradiol-synthesizing enzyme aromatase (CYP19A1); these changes require inputs from the afferent song control nucleus HVC (used as a proper name) and steroid receptor activation within HVC; local coactivation of androgen and estrogen receptors (ARs and ERs, respectively) within HVC, but not RA, is sufficient to cause neuronal changes in RA; activation of ARs in RA is also permissive. Using bilateral local intracerebral hormone-receptor blockade, we found that ARs and ERs in the song-control nucleus HVC mediate systemic testosterone-induced changes in song stereotypy but not rate. This novel transsynaptic effect of gonadal steroids on activity and morphology of RA neurons is part of a concerted change in key premotor nuclei, enabling stereotyped song.

In numerous species of birds, individuals or species that sing larger numbers of song types have larger song control nuclei in their brains. The direction of the cause and effect relationship between the complexity of song behavior and brain space is unknown, however. The hypothesis that birds that learn large song repertoires develop large song nuclei was therefore tested with a songbird, the marsh wren (Cistothorus palustris). Males were hand-reared and tutored in the laboratory with either small (n = 8 males heard 5 song types) or large (n = 8 males heard 45 song types) repertoires. When the birds were adults, the number of song types each male sang was first determined, and then the volume and certain cellular attributes of the song nuclei HVC and RA were measured. The two groups of wrens showed large behavioral differences in the size of their learned song repertoires, but did not differ in either the volumes of HVC and RA or in neuronal size, number, or density within these nuclei. These results suggest that the relationship between behavioral song complexity and brain space found in this and other species develops largely independently of early song learning experience and the later production of those songs.

There is pronounced seasonal plasticity in the morphology of the neural circuits that regulate song behavior in adult songbirds, primarily in response to changes in plasma testosterone (T) levels. Most song nuclei have androgen receptors. Afferent input from the telencephalic nucleus HVc (also known as the ''high vocal center'') is necessary for seasonal growth of the direct efferent target nuclei RA and area X. We asked here whether T-stimulated growth of HVc is sufficient to induce growth of its efferent nuclei. Intracerebral T implants were placed unilaterally near HVc or RA in photosensitive adult male white-crowned sparrows for one month. The T implant near HVc produced significant growth of the ipsilateral (but not contralateral) HVc, RA, and area X, and increased neuronal number in the ipsilateral HVc. The T implant near RA did not produce selective growth of ipsilateral RA, HVc, or area X. Intracerebral T implants did not elevate plasma T levels, nor did they stimulate growth of two peripheral androgen sensitive targets, the syrinx and the cloacal protuberance. These results suggest that seasonal growth of the adult song circuits results from T acting directly on HVc, which then stimulates the growth of RA and area X transynaptically.

Photoperiod and hormonal cues drive dramatic seasonal changes in structure and function of the avian song control system. Little is known, however, about the patterns of gene expression associated with seasonal changes. Here we address this issue by altering the hormonal and photoperiodic conditions in seasonally-breeding Gambel's white-crowned sparrows and extracting RNA from the telencephalic song control nuclei HVC and RA across multiple time points that capture different stages of growth and regression. We chose HVC and RA because while both nuclei change in volume across seasons, the cellular mechanisms underlying these changes differ. We thus hypothesized that different genes would be expressed between HVC and RA. We tested this by using the extracted RNA to perform a cDNA microarray hybridization developed by the SoNG initiative. We then validated these results using qRT-PCR. We found that 363 genes varied by more than 1.5 fold (>log2 0.585) in expression in HVC and/or RA. Supporting our hypothesis, only 59 of these 363 genes were found to vary in both nuclei, while 132 gene expression changes were HVC specific and 172 were RA specific. We then assigned many of these genes to functional categories relevant to the different mechanisms underlying seasonal change in HVC and RA, including neurogenesis, apoptosis, cell growth, dendrite arborization and axonal growth, angiogenesis, endocrinology, growth factors, and electrophysiology. This revealed categorical differences in the kinds of genes regulated in HVC and RA. These results show that different molecular programs underlie seasonal changes in HVC and RA, and that gene expression is time specific across different reproductive conditions. Our results provide insights into the complex molecular pathways that underlie adult neural plasticity.

Seasonal plasticity in the morphology of telencephalic nuclei that control song behavior has been reported for diverse species of songbirds. The only published report of a lack of seasonal changes in the song nuclei of a seasonally breeding bird is that of Baker et al. in the Nuttall's subspecies of white‐crowned sparrow (Zonotrichia leucophrys nuttalli) . In this study, they brought wild birds into the laboratory and exposed them to either “summer” or “winter” photoperiods. Previous studies have shown that exposing wild‐caught white‐crowned sparrows to long‐day photoperiods in the laboratory may not induce circulating concentrations of testosterone (T) as high as those seen in wild breeding birds. Changes in circulating T are primarily responsible for the seasonal morphological changes in the song nuclei. To determine whether there is seasonal plasticity of the song system in this subspecies, we measured circulating T, morphological attributes of the song nuclei, and song behavior in wild Nuttall's white‐crowned sparrows during the spring and fall. Testis size and circulating T concentrations were greater in spring than fall birds. The absolute volumes of the song nuclei HVc, RA, and Area X, and their volumes relative to those of either the total telencephalon or three thalamic nonsong nuclei, were significantly greater in the spring than fall sparrows. Song behavior also changed seasonally; fall birds sang shorter songs than did spring birds. These results show that there is seasonal plasticity of the song system in wild Nuttall's white‐crowned sparrows. Seasonal plasticity can now be regarded as a common feature of the seasonally breeding songbirds studied thus far. © 1998 John Wiley & Sons, Inc. J Neurobiol 34: 69–82, 1998

The neural circuits that regulate song behavior in adult songbirds undergo pronounced seasonal changes in morphology, primarily in response to changes in plasma testosterone (T). Most song nuclei have T receptors. We asked whether seasonal growth and maintenance of nuclei within these circuits are direct responses to the effects of T or its metabolites or are mediated indirectly via the effects of T on afferent nuclei. Photosensitive white-crowned sparrows were exposed to one of three treatments. (1) The neostriatal nucleus HVc (also known as the "high vocal center") was lesioned unilaterally, and the birds were exposed to long-day (LD) photoperiods and breeding levels of T for 30 d. (2) Birds were exposed to LD plus T (LDϩT) for 30 d; then HVc was lesioned, and the birds were killed after an additional 30 d exposure to LDϩT. (3) HVc was lesioned, and the sparrows were housed on short-day (SD) photoperiods in the absence of T treatment for 30 d. In both LDϩT groups, the direct efferent targets of HVc, the robust nucleus of the archistriatum (RA) and area X, were smaller ipsilateral to the lesion. The lesion did not prevent growth of the hypoglossal motor nucleus, which does not receive direct afferent input from HVc. RA and area X were also smaller ipsilateral to the lesion in the SD birds. These results indicate that afferent input is required both for the growth of adult song circuits in response to typical breeding photoperiod and hormone conditions and for the maintenance of efferent nuclei in either their regressed or enlarged states.

No abstract

In songbirds, neurons that regulate learned song behavior undergo extensive seasonal plasticity in their number and size in relation to the bird's reproductive status. Seasonal plasticity of these brain regions is primarily regulated by changes in circulating concentrations of testosterone. Androgen receptors are present in all of the major song nuclei, but it is unknown whether levels of androgen receptor mRNA in the telencephalic song regions HVC, the robust nucleus of the arcopallium, and the lateral magnocellular nucleus of the anterior nidopallium change as a function of season in whitecrowned sparrows. To determine whether seasonal changes in levels of androgen receptor mRNA are specific to the song control system, we also measured levels of androgen receptor mRNA in a limbic nucleus, the the lateral division of the bed nucleus of the stria terminalis (the lateral division of the bed nucleus of the stria terminalis). We found that levels of androgen receptor mRNA were higher in HVC and the lateral division of the bed nucleus of the stria terminalis of birds in the breeding-condition compared with the nonbreeding-condition; however, we observed no seasonal differences in levels of androgen receptor mRNA in either the the robust nucleus of the arcopallium or the lateral magnocellular nucleus of the anterior nidopallium. These results are consistent with previous observations that seasonal plasticity of the song nuclei results fromtestosteronacting directly on HVC, which then exerts transsynaptic trophic effects on its efferent targets. The seasonal change in the expression of androgen receptor in HVC may be one component of the cellular mechanisms underlying androgenic effects on seasonal plasticity of the song control nuclei.