Subdivisions of the auditory midbrain (n. mesencephalicus lateralis, pars dorsalis) in zebra finches using calcium-binding protein immunocytochemistry.

The midbrain nucleus mesencephalicus lateralis pars dorsalis (MLd) is thought to be the avian homologue of the central nucleus of the mammalian inferior colliculus. As such, it is a major relay in the ascending auditory pathway of all birds and in songbirds mediates the auditory feedback necessary for the learning and maintenance of song. To clarify the organization of MLd, we applied three calcium binding protein antibodies to tissue sections from the brains of adult male and female zebra finches. The staining patterns resulting from the application of parvalbumin, calbindin and calretinin antibodies differed from each other and in different parts of the nucleus. Parvalbumin-like immunoreactivity was distributed throughout the whole nucleus, as defined by the totality of the terminations of brainstem auditory afferents; in other words parvalbumin-like immunoreactivity defines the boundaries of MLd. Staining patterns of parvalbumin, calbindin and calretinin defined two regions of MLd: inner (MLd.I) and outer (MLd.O). MLd.O largely surrounds MLd.I and is distinct from the surrounding intercollicular nucleus. Unlike the case in some non-songbirds, however, the two MLd regions do not correspond to the terminal zones of the projections of the brainstem auditory nuclei angularis and laminaris, which have been found to overlap substantially throughout the nucleus in zebra finches.

Trigeminal and spinal dorsal horn (dis)continuity and avian evolution.

The organization of the dorsal horn in the avian spinal cord differs in different species. For instance, in pigeons and doves, cranes, cuckoos, songbirds, ratites and tinamous, the dorsal horn is organized in laminar fashion, such that laminae II and III are sandwiched between lamina I dorsally and lamina IV ventrally, as they are in mammals and other nonavian amniotes. In most other avian species, including chickens, however, the organization of the dorsal horn is not strictly laminar, in that the structures homologous to laminae II and III lie side by side rather than in dorsoventral order. Because spinal and trigeminal dorsal horns are generally thought to be continuous, the question arises as to the organization of the trigeminal dorsal horn in these species. We examined this question in chickens, first by defining II and III of trigeminal and spinal dorsal horns using calcium-binding protein immunohistochemistry, and second by determining the caudal extent of the projections of the three branches of the trigeminal nerve using injections of cholera toxin B chain. It was found (1) that the trigeminal dorsal horn and the spinal dorsal horn of the first 2 cervical segments are organized in laminar fashion, but further caudally, II and III in the spinal dorsal horn gradually come to be arranged side by side and (2) that the descending trigeminal tract terminates no further caudal than the 3rd spinal segment. Therefore, unlike spinal nerves, trigeminal nerve branches do not project to II and III, once these cease to be organized in laminar fashion. These findings imply some kind or organizational discontinuity of trigeminal and spinal dorsal horns in the chicken and perhaps in other species with a side-by-side arrangement of II and III. It has also been suggested that the condition in which the spinal dorsal horn structures homologous to laminae II and II lie side by side may define a novel clade of birds. This suggestion was reexamined within the context of a modern phylogenetic framework based on 32 kb of nuclear DNA, and using a parsimony reconstruction of dorsal horn character states. The original suggestion of a novel clade was not supported. Instead, it appears that the side-by-side condition evolved very early in the radiation of Aves and that independent reversion to a laminar dorsal horn condition has evolved at least 4-5 times.

Connections of the auditory brainstem in a songbird, Taeniopygia guttata. II. Projections of nucleus angularis and nucleus laminaris to the superior olive and lateral lemniscal nuclei.

Three nuclei of the lateral lemniscus are present in the zebra finch, ventral (LLV), intermediate (LLI), and dorsal (LLD). LLV is separate from the superior olive (OS): it lies closer to the spinal lemniscus and extends much further rostrally around the pontine periphery. LLI extends from a caudal position ventrolateral to the principal sensory trigeminal nucleus (LLIc) to a rostral position medial to the ventrolateral parabrachial nucleus (LLIr). LLD consists of posterior (LLDp) and anterior (LLDa) parts, which are largely coextensive rostrocaudally, although LLDa lies medial to LLDp. All nuclei are identifiable on the basis of cytochrome oxidase activity. The cochlear nucleus angularis (NA) and the third-order nucleus laminaris (NL) project on OS predominantly ipsilaterally, on LLV and LLI predominantly contralaterally, and on LLD contralaterally only. The NA projections are heavier than those of NL and differ from them primarily in their terminations within LLD: NA projects to LLDp, whereas NL projects to LLDa. In this the projections are similar to those in the barn owl (Takahashi and Konishi [1988] J Comp Neurol 274:212-238), in which time and intensity pathways remain separate as far as the central nucleus of the inferior colliculus (MLd). In contrast, in the zebra finch, although NA and NL projections remain separate within LLD, the projections of LLDa and LLDp become intermixed within MLd (Wild et al., J Comp Neurol, this issue), consistent with the intermixing of the direct NA and NL projections to MLd (Krützfeldt et al., J Comp Neurol, this issue).

Connections of the auditory brainstem in a songbird, Taeniopygia guttata. I. Projections of nucleus angularis and nucleus laminaris to the auditory torus.

Auditory information is important for social and reproductive behaviors in birds generally, but is crucial for oscine species (songbirds), in particular because in these species auditory feedback ensures the learning and accurate maintenance of song. While there is considerable information on the auditory projections through the forebrain of songbirds, there is no information available for projections through the brainstem. At the latter levels the prevalent model of auditory processing in birds derives from an auditory specialist, the barn owl, which uses time and intensity parameters to compute the location of sounds in space, but whether the auditory brainstem of songbirds is similarly functionally organized is unknown. To examine the songbird auditory brainstem we charted the projections of the cochlear nuclei angularis (NA) and magnocellularis (NM) and the third-order nucleus laminaris (NL) in zebra finches using standard tract-tracing techniques. As in other avian species, the projections of NM were found to be confined to NL, and NL and NA provided the ascending projections. Here we report on differential projections of NA and NL to the torus semicircularis, known in birds as nucleus mesencephalicus lateralis, pars dorsalis (MLd), and in mammals as the central nucleus of the inferior colliculus (ICc). Unlike the case in nonsongbirds, the projections of NA and NL to MLd in the zebra finch showed substantial overlap, in agreement with the projections of the cochlear nuclei to the ICc in mammals. This organization could suggest that the "what" of auditory stimuli is as important as "where."

Connections of the auditory brainstem in a songbird, Taeniopygia guttata. III. Projections of the superior olive and lateral lemniscal nuclei.

Sequential to companion articles that report the projections of the cochlear nucleus angularis (NA) and the third-order nucleus laminaris (NL) to the central nucleus of the inferior colliculus (MLd) and to the superior olive (OS) and lateral lemniscal nuclei (LLV, LLI, and LLD) (Krützfeldt et al., J Comp Neurol, this issue), we here describe the projections of the latter group of nuclei using standard tract-tracing methods. OS projects on LLV and both have further ascending projections on LLI, LLD, and MLd. LLV also provides auditory input to the song system, via nucleus uvaeformis, and to the thalamo-telencephalic auditory system, via nucleus ovoidalis (Ov), thus bypassing MLd. The two divisions of LLD (LLDa and LLDp) project across the midline via the commissure of Probst each to innervate the homologous contralateral nucleus and MLd. Both, particularly LLDp, also project on Ov. Injections in LLD and LLV resulted in anterograde labeling of caudal nucleus basorostralis (Bas) in the frontal telencephalon, but retrograde tracing so far suggests that only LLI is a real source of this projection (Wild and Farabaugh [1996] J Comp Neurol 365:306-328). OS and LLV also have descending projections on the ipsilateral NA, NM, and NL, and LLV also projects on OS. The ascending inputs to MLd and more rostral nuclei may contribute importantly to mechanisms of auditory pattern (song) recognition. Consistent with previous studies, some of the descending projections may be inhibitory.

Definition and novel connections of the entopallium in the pigeon (Columba livia).

The avian entopallium (E) is the major thalamorecipient zone, within the telencephalon, of the tectofugal visual system. Because of discrepancies concerning the structure of this nuclear mass in pigeons, and in light of recent evidence concerning entopallial projections in other avian species, we here redefine and chart some novel entopallial projections in the pigeon by using a combination of cytochrome oxidase (CO) activity, calcium binding protein immunohistochemistry (CBPi), normal histology, and tract tracing. We show that 1) E is defined by the accurate overlap of CO activity and the dense terminations of thalamic (rotundal) efferents; 2) the perientopallium (Ep), E's overlying belt region, receives a relatively sparse rotundal input and is a major source of projections to wider regions of the hemisphere; and 3) E can be subdivided into internal (Ei) and external (Ex) portions on the basis of normal histology, CBPi, and differential projections. Thus, Ei, but not Ex, makes a reciprocal connection with a distinct nucleus in the ventrolateral mesopallium and is a major source of projections to the lateral striatum. These findings suggest the necessity for a revision of the original proposal of a strictly serial flow of visual information through the entopallial complex and further regions of the hemisphere and also require a modification of the long-standing view that E is comparable to only one specific lamina (IV) of extrastriate visual cortex of mammals. Rather, E appears to be composed of a variety of neuronal types possibly equivalent to those in several neocortical laminae.

Definition and connections of the entopallium in the zebra finch (Taeniopygia guttata).

In birds the entopallium (formerly known as the core region of ectostriatum) is the major thalamorecipient zone, within the telencephalon, of the tectofugal visual system. Here we sought to redefine the entopallium in the zebra finch, particularly with respect to a laterally adjacent zone, known as the perientopallium (formerly known as the periectostriatal belt), and to determine its projections. We show that the entopallium can be defined by the almost complete overlap of dense terminations of thalamic rotundal afferents and intense cytochrome oxidase activity and parvalbumin immunoreactivity. The perientopallium, on the other hand, can be defined by relatively sparse projections from nucleus rotundus, a calretinin-positive plexus of nerve fibers, and weak cytochrome oxidase activity and parvalbumin immunoreactivity. Within the entopallium, medial and lateral parts can be distinguished on the basis of cell packing density, differential patterns of parvalbumin immunoreactivity and cytochrome oxidase activity, and different projections. We show that the entopallium projects laterally and diffusely to the perientopallium and nidopallium (formerly the neostriatum) and specifically and densely to a teardrop-shaped nucleus in the ventrolateral mesopallium (formerly known as the hyperstriatum ventrale), here called MVL (abbreviation used as a proper name). This latter projection arises predominantly from medial parts of the entopallium, which also receives a reciprocal projection from MVL, and projects to the lateral striatum. These findings suggest that the entopallium can be divided into medial and lateral parts having different functions, one of which is to provide for an extratelencephalic outflow from the medial part, via the lateral striatum. The findings also challenge the idea that informational flow through the various stations of the telencephalic tectofugal visual system is largely sequential and, together with findings in the chicken (Alpar and Tömböl), suggest instead that further substantial projections to telencephalic visual areas in birds can arise independently from both E (entopallium) and Ep (perientopallial belt).