Subdivisions of the mesencephalon and isthmus in the lizard Gekko gecko as revealed by ChAT immunohistochemistry.

The distribution of cholinergic cell bodies and fibers was examined in the mesencephalon and isthmus of Gekko gecko. Distinct groups with prominent labeled cells were observed in the cranial nerve motor nuclei and isthmic nuclei, and weak labeled cell bodies and fibers were observed in the mesencephalic nucleus of the trigeminal nerve and the central nucleus of the torus semicircularis. After discussing the topological relationships within the tectum and isthmus, we unify the nomenclature of the caudal deep mesencephalic nucleus in lizards and the rostral magnocellular nucleus isthmi in turtles that is similar in terms of the preisthmic position, nontopographic connections with the tectum, and the same midbrain origin to the magnocellular preisthmic nucleus in birds, and may be homologous to the superficial cuneiform nucleus in mammals. None of them belong to the cholinergic nucleus isthmi, as the latter has isthmus origin and topographic reciprocal connections with the tectum. We also discuss the origin and intrinsic function of the inner longitudinal tract of the thick ChAT-ir fibers that course through the mesencephalon and diencephalon. We review the subdivisions of the mesencephalon and isthmus of Gekko gecko as revealed by ChAT immunohistochemistry, as well as the limits of the diencephalo-mesencephalic, mesencephalic-isthmo, and isthmo-rhombocephalic by the ChAT-ir cell- and fiber-poor distribution, and discuss the caudal limit of the isthmus. Our research on the subdivisions of the mesencephalon and isthmus in G. gecko as revealed by ChAT immunohistochemistry will serve as the neuroanatomical basis for subsequent relevant studies of Gekko gecko.

Calcium-binding protein immunoreactivity characterizes the auditory system of Gekko gecko.

Geckos use vocalizations for intraspecific communication, but little is known about the organization of their central auditory system. We therefore used antibodies against the calcium-binding proteins calretinin (CR), parvalbumin (PV), and calbindin-D28k (CB) to characterize the gecko auditory system. We also examined expression of both glutamic acid decarboxlase (GAD) and synaptic vesicle protein (SV2). Western blots showed that these antibodies are specific to gecko brain. All three calcium-binding proteins were expressed in the auditory nerve, and CR immunoreactivity labeled the first-order nuclei and delineated the terminal fields associated with the ascending projections from the first-order auditory nuclei. PV expression characterized the superior olivary nuclei, whereas GAD immunoreactivity characterized many neurons in the nucleus of the lateral lemniscus and some neurons in the torus semicircularis. In the auditory midbrain, the distribution of CR, PV, and CB characterized divisions within the central nucleus of the torus semicircularis. All three calcium-binding proteins were expressed in nucleus medialis of the thalamus. These expression patterns are similar to those described for other vertebrates.

Development of N-methyl-D-aspartate receptor subunits in avian auditory brainstem.

N-methyl-D-aspartate (NMDA) receptor subunit-specific probes were used to characterize developmental changes in the distribution of excitatory amino acid receptors in the chicken's auditory brainstem nuclei. Although NR1 subunit expression does not change greatly during the development of the cochlear nuclei in the chicken (Tang and Carr [2004] Hear. Res 191:79-89), there are significant developmental changes in NR2 subunit expression. We used in situ hybridization against NR1, NR2A, NR2B, NR2C, and NR2D to compare NR1 and NR2 expression during development. All five NMDA subunits were expressed in the auditory brainstem before embryonic day (E) 10, when electrical activity and synaptic responses appear in the nucleus magnocellularis (NM) and the nucleus laminaris (NL). At this time, the dominant form of the receptor appeared to contain NR1 and NR2B. NR2A appeared to replace NR2B by E14, a time that coincides with synaptic refinement and evoked auditory responses. NR2C did not change greatly during auditory development, whereas NR2D increased from E10 and remained at fairly high levels into adulthood. Thus changes in NMDA NR2 receptor subunits may contribute to the development of auditory brainstem responses in the chick.

Development of NMDA R1 expression in chicken auditory brainstem.

NMDA receptor subunit 1 (NR1) expression in the chicken cochlear nuclei was examined using immunohistochemistry and quantitative Western blots. An antibody raised in mouse against a highly conserved domain of NR1 recognized the same 115 kDa protein band in chicken brain. Quantitative Western blotting of cochlear nucleus protein showed no significant change in NR1 expression from E18 to adult. The nucleus angularis (NA) initiated NR1 expression before E12 that became more prominent after hatching. NR1-ir first appeared in the nucleus magnocellularis (NM) and nucleus laminaris (NL) at E10. From E12 to E19, NM exhibited a gradient in NR1 expression with medial, higher best frequency cell bodies being more immunoreactive than lateral, lower best frequency cell bodies. This gradient disappeared by E20. The distribution of NR1 in NL also changed during development. NR1 label was present in NL cell bodies between E10 and E13. From E14 onwards, NR1-ir characterized both cell bodies and neuropil. After hatching, NR1-ir levels were higher in NL than NM. The superior olive first expressed NR1 at E12. Neuropil staining was more intense than cell bodies until after hatching. In contrast to the functional decrease observed in mammals and chick, NR1-ir expression remained high in the chicken auditory brainstem into adulthood. Both chickens and rodents retain high levels of NR-1.

Contact call-driven zenk mRNA expression in the brain of the budgerigar (Melopsittacus undulatus).

Contact call-driven zenk (zif268, egr1, NGF1A, Krox 24) mRNA expression was mapped with in situ hybridization histochemistry in a vocal learning parrot, the budgerigar (M. undulatus). Relative to controls, call stimulation induced high zenk mRNA expression in all auditory areas including those closely associated with the vocal system within the anterior forebrain (Brauth et al. (2001) J. Comp. Neurol. 432, 481; (2002) Learn. Memory 9, 76). Thus there is a high correspondence between the distributions of neurons exhibiting contact call-driven zenk protein and mRNA expression in budgerigars. Field L2a, an area reported previously to express only perinucleolar zenk protein localization (Brauth et al. (2002) Learn. Memory 9, 76) also showed zenk mRNA expression.