Bone conduction pathways confer directional cues to salamanders.

Sound and vibration are generated by mechanical disturbances within the environment, and the ability to detect and localize these acoustic cues is generally important for survival, as suggested by the early emergence of inherently directional otolithic ears in vertebrate evolutionary history. However, fossil evidence indicates that the water-adapted ear of early terrestrial tetrapods lacked specialized peripheral structures to transduce sound pressure (e.g. tympana). Therefore, early terrestrial hearing should have required nontympanic (or extratympanic) mechanisms for sound detection and localization. Here, we used atympanate salamanders to investigate the efficacy of extratympanic pathways to support directional hearing in air. We assessed peripheral encoding of directional acoustic information using directionally masked auditory brainstem response recordings. We used laser Doppler vibrometry to measure the velocity of sound pressure-induced head vibrations as a key extratympanic mechanism for aerial sound reception in atympanate species. We found that sound generates head vibrations that vary with the angle of the incident sound. This extratympanic pathway for hearing supports a figure-eight pattern of directional auditory sensitivity to airborne sound in the absence of a pressure-transducing tympanic ear.

Seismic sensitivity and bone conduction mechanisms enable extratympanic hearing in salamanders.

The tympanic middle ear is an adaptive sensory novelty that evolved multiple times in all the major terrestrial tetrapod groups to overcome the impedance mismatch generated when aerial sound encounters the air-skin boundary. Many extant tetrapod species have lost their tympanic middle ears, yet they retain the ability to detect airborne sound. In the absence of a functional tympanic ear, extratympanic hearing may occur via the resonant qualities of air-filled body cavities, sensitivity to seismic vibration, and/or bone conduction pathways to transmit sound from the environment to the ear. We used auditory brainstem response recording and laser vibrometry to assess the contributions of these extratympanic pathways for airborne sound in atympanic salamanders. We measured auditory sensitivity thresholds in eight species and observed sensitivity to low-frequency sound and vibration from 0.05-1.2 kHz and 0.02-1.2 kHz, respectively. We determined that sensitivity to airborne sound is not facilitated by the vibrational responsiveness of the lungs or mouth cavity. We further observed that, although seismic sensitivity probably contributes to sound detection under naturalistic scenarios, airborne sound stimuli presented under experimental conditions did not produce vibrations detectable to the salamander ear. Instead, threshold-level sound pressure is sufficient to generate translational movements in the salamander head, and these sound-induced head vibrations are detectable by the acoustic sensors of the inner ear. This extratympanic hearing mechanism mediates low-frequency sensitivity in vertebrate ears that are unspecialized for the detection of aerial sound pressure, and may represent a common mechanism for terrestrial hearing across atympanic tetrapods.

Evolution and development of time coding systems.

Precise temporal coding is a hallmark of both the electrosensory and auditory systems. Selective pressures to improve accuracy or encode more rapid changes have produced a suite of convergent physiological and morphological features that contribute to temporal coding. Comparative studies of temporal coding can also point to shared computational strategies, and suggest how selection might act to improve coding.

Expression of the Kv3.1 potassium channel in the avian auditory brainstem.

The Shaw-like potassium channel Kv3.1, a delayed rectifier with a high threshold of activation, is expressed in the time coding nuclei of the bird auditory brainstem. In both barn owls and chickens, Kv3.1 mRNA was expressed in the cochlear nucleus magnocellularis (NM) and the nucleus laminaris (NL). Western blot analysis showed that an antibody raised against the synthetic peptide sequence of rat Kv3.1 (rKv3.1) specifically recognized the same 92 kDa protein bands in both rat and chicken synaptosomal preparations. Immunohistochemical analyses using this anti-rKv3.1 antibody revealed a prominent gradient in Kv3.1 immunoreactivity along the tonotopic axis of the barn owl NM and NL and a less prominent gradient in the chicken. The precise localization of the Kv3.1 immunoproduct was resolved by electron microscopy. In both the owl and the chicken, Kv3.1 was targeted postsynaptically in NM and NL. The major difference in localization of Kv3.1 protein between the two birds was the expression of Kv3.1 in the NM axons and terminals in the region of the barn owl NL. This location of Kv3.1 channels supports its postulated function in reducing the width of action potentials as they invade the presynaptic terminal. The presynaptic localization may be a specialization for enabling neurons in owl NM to transmit high-frequency temporal information with little jitter.

Transforming growth factor beta from multiple myeloma cells inhibits proliferation and IL-2 responsiveness in T lymphocytes.

Multiple myeloma (MM) is a cancer of plasma cells, characterized by profound suppression of host immune responses. Here we show that MM cell lines significantly suppress the proliferation, blasting, response to interleukin-2 (IL-2), and expression of CD25 by concanavalin A (Con A)-activated or allostimulated peripheral blood T lymphocytes. T cells arrest in the G1 stage of the cell cycle, and do not enter the IL-2 autocrine growth pathway. T cell inhibition was mediated by a soluble factor. MM cell lines did not produce IL-10 but did produce large amounts of transforming growth factor beta1 (TGF-beta1). T cells were assessed for their ability to respond to IL-2 when co-cultured with MM cells in the presence or absence of the TGF-beta inhibitor, TGF-beta latency-associated peptide (LAP). MM cells suppressed IL-2 responses but this inhibition was completely reversed by TGF-beta LAP. A CD25-, IL-2-dependent blast cell line was not inhibited by MM cells or rhTGF-beta, confirming the specificity of the inhibition mechanism for the IL-2 autocrine growth pathway. We conclude that MM cells suppress T cells in their entry into the autocrine IL-2/CD25 pathway and in response to IL-2, and that TGF-beta has a significant role to play.

The cytoarchitecture of the nucleus angularis of the barn owl (Tyto alba).

The cochlear nucleus angularis (NA) of the barn owl (Tyto alba) was analyzed using Golgi, Nissl, and tract tracing techniques. NA forms a column of cells in the dorsolateral brainstem that partly overlaps with, and is rostral and lateral to, the cochlear nucleus magnocellularis (NM). Highest best frequencies are mapped in lateral NA (NAl), intermediate in medial NA (NAm), and lowest in the foot region (NAf). Cell density followed the tonotopic axis and decreased with decreasing best frequency. NA contained four major cell classes: planar, radiate, vertical, and stubby. Planar and radiate classes were further subdivided into bipolar and multipolar types according to their number of primary dendrites. Planar neurons were confined to an isofrequency band, whereas radiate neurons had dendrites that could extend across an isofrequency band. Vertical cells had long dendrites oriented perpendicularly to isofrequency bands. Stubby cells were the most numerous and were confined to an isofrequency band because of their short dendrites. Neurons in each of these four classes projected to the inferior colliculus and dorsal nucleus of the lateral lemniscus.

Central projections of auditory nerve fibers in the barn owl.

The central projections of the auditory nerve were examined in the barn owl. Each auditory nerve fiber enters the brain and divides to terminate in both the cochlear nucleus angularis and the cochlear nucleus magnocellularis. This division parallels a functional division into intensity and time coding in the auditory system. The lateral branch of the auditory nerve innervates the nucleus angularis and gives rise to a major and a minor terminal field. The terminals range in size and shape from small boutons to large irregular boutons with thorn-like appendages. The medial branch of the auditory nerve conveys phase information to the cells of the nucleus magnocellularis via large axosomatic endings or end bulbs of Held. Each medial branch divides to form 3-6 end bulbs along the rostrocaudal orientation of a single tonotopic band, and each magnocellular neuron receives 1-4 end bulbs. The end bulb envelops the postsynaptic cell body and forms large numbers of synapses. The auditory nerve profiles contain round clear vesicles and form punctate asymmetric synapses on both somatic spines and the cell body.

Organization of the nucleus magnocellularis and the nucleus laminaris in the barn owl: encoding and measuring interaural time differences.

The circuit from the cochlear nucleus magnocellularis to the nucleus laminaris supports the encoding and measurement of interaural time differences in the auditory brainstem. Specializations for the encoding of temporal information include the few and/or short dendrites and thick axons of the magnocellular and laminaris neurons, and the high degree of convergence in the circuit. Magnocellular cells have large cell bodies covered with somatic spines. The cells have few dendrites, and the number of dendrites decreases from low to high best frequency regions of the nucleus. Magnocellular neurons receive both auditory nerve terminals and GABAergic terminals with symmetric synapses and terminals filled with pleomorphic vesicles. The axonal projections of magnocellular neurons to the nucleus laminaris form maps of interaural time difference. About 100 magnocellular afferents from each side converge on each laminaris neuron, and the terminals from each side do not occupy separate domains on the cell. These terminals form punctate asymmetric synapses on both the dendrites and the cell bodies of laminaris neurons. Laminaris neurons also receive GABAergic terminals which form symmetric synapses. Laminaris neurons have oval cell bodies covered with very short dendrites. The cells in the low best frequency region of the nucleus laminaris have longer dendrites.

Low-frequency pathway in the barn owl's auditory brainstem.

The cytology of the nucleus magnocellularis and the nucleus laminaris in the barn owl, as well as the axonal pathways connecting them, were studied. The interest was focussed on those regions of both nuclei coding the low-frequency end of the tonotopic spectrum (below approximately 2 kHz) because many previous reports on a variety of bird species had indicated significant differences to higher frequencies, both in morphology and physiology. Standard light- and electron microscopy, as well as immunocytochemistry and tract-tracing techniques, were used. The nucleus magnocellularis contains a distinct stellate cell type in the low-frequency region, in addition to neurons classified as a small version of the principal cell. In the nucleus laminaris, two cell types were characterized as distinct to the low-frequency region: stellate neurons with long, smooth dendrites, and multipolar neurons with thick, spiny dendrites. The low-frequency projections from the nucleus magnocellularis showed two terminal fields in the nucleus laminaris: one containing a rough tonotopic representation and a second one where all low-frequency projections converged. In addition, the anatomical basis for delay lines, which are known to play an important role in the coding of interaural time differences at higher frequencies, was not observed. The morphological differences observed at low frequencies in both nuclei, compared to the well-studied higher-frequency regions, may reflect inherent limitations to the accuracy in the processing of interaural phase disparities at low frequencies.

Development of the time coding pathways in the auditory brainstem of the barn owl.

The barn owl's head grows after hatching, causing interaural distances to more than double in the first 3 weeks posthatch. These changes expose the bird to a constantly increasing range of interaural time cues. We have used Golgi and ultrastructural techniques to analyze the development of the connections and cell types of the nucleus magnocellularis (NM) and the nucleus laminaris (NL) with reference to the growth of the head. The time coding circuit is formed but immature at the time of hatching. In the month posthatch, the auditory nerve projection to the NM matures, and appears adult-like by posthatch day (P)21. NM neurons show a late growth of permanent dendrites starting at P6. Over the first month, these dendrites change in length and number, depending upon rostrocaudal position, to establish the adult pattern in which high best frequency neurons have few or no dendrites. These changes are not complete by P21, when NM neurons still have more dendrites than in the adult owl. The neurons of NL have many short dendrites before hatching. Their number is greatly reduced by P6, and then does not change during later development. Like NM neurons, NL neurons and dendrites grow in the first month posthatch, and at P21, NL dendrites are longer than those in the adult owl. Thus, the auditory brainstem circuits grow in the first month after hatching, but are not yet mature at the time the head reaches its adult size.

Choline acetyltransferase-immunoreactive cochlear efferent neurons in the chick auditory brainstem.

Cholinergic neurons in the chick auditory brainstem were studied with the aid of an antiserum to choline acetyltransferase (ChAT), the biosynthetic enzyme for acetylcholine. ChAT-immunoreactive (ChAT-I) neurons were found in a ventrolateral and a dorsomedial cell group. The ventrolateral group is a rostrocaudally directed column of cells that surround the superior olive (SO), are ventromedial to the ventral facial nucleus (VIIv), and are lateral to the nucleus pontis lateralis (PL) as far rostrally as the nucleus subceruleus ventralis. Cells in the dorsomedial group were found in the pontine reticular formation medial to the dorsal facial nucleus and lateral to the abducens nerve root. Occasionally, small ChAT-I cells were found in the crossed dorsal cochlear tract and in the medial vestibular nucleus near the dorsal border of the caudal nucleus magnocellularis (NM). No ChAT-I neurons or fibers were observed in NM, nucleus angularis, nucleus laminaris, in the nuclei of the lateral lemniscus, or in the nucleus mesencephalicus lateralis pars dorsalis. To determine which cholinergic neurons project to the cochlea, a double-labeling technique was used combining ChAT-I and the retrograde transport of biotinylated dextran amine (BDA) from the inner ear. Double-labeled cells were found bilaterally in both the ventrolateral and dorsomedial cell groups, with the exception of large ChAT-I cells dorsal to the SO, which do not appear to project to the cochlea. Cholinergic cells that project to the cochlea were classified into three morphological groups: multipolar, elongate, and round-to-oval. Both the ventrolateral and the dorsomedial cell groups appear to have a mixture of these different cell types. The average somal area of cholinergic cochlear efferents was 246 microns 2. Only about 70% of the cochlear efferent neurons, however, are cholinergic.

A Golgi study of the cell types of the dorsal torus semicircularis of the electric fish Eigenmannia: functional and morphological diversity in the midbrain.

The dorsal torus semicircularis (torus) of the gymnotiform fish Eigenmannia was examined in Golgi-impregnated material. These results were correlated with those of a previous HRP study which used retrograde labelling techniques to identify the efferent cell types of the torus (Carr et al., '81, J. Comp Neurol. 203:649-670). The torus is a laminated midbrain nucleus of the electrosensory system. It receives somatotopically ordered electrosensory input from the medulla and caudal lobe of the cerebellum, proprioceptive input from the descending nucleus of the trigeminal nerve, and input from the optic tectum. The torus projects to the nucleus praeeminentialis, the optic tectum, nucleus electro-sensorius, parts of the central posterior thalamus, the pretectum, the lateral mesencephalic reticular formation (LMRA), the reticular formation, and the inferior olive. The torus has 12 laminae and 48 cell types by Golgi criteria. There are three major orientations to the dendritic fields of the toral neurons: purely horizontal neurons with dendrites confined to a single lamina, multipolar neurons whose dendrites often do not respect laminar boundaries, and vertical cells with dendrites that travel in the vertical bundles of dendrites and axons which pierce the torus at regular intervals. There are four major groups of vertically oriented neurons. The first has a predominantly horizontal dendritic tree with one or two vertical dendrites which connect the cell to a distant lamina. The second consists of "U"-shaped neurons with a horizontal arbor and two major dendrites which ascend in adjacent vertical bundles. The third group is made up of bilaminar neurons which receive input from two vertically separated dendritic arbors, and the fourth group is purely vertical in orientation. A group of four tegmental cell types in the LMRA also send their dendrites into the efferent tracts of the torus, and into lamina IX. The torus is similar in complexity and number of cell types to the mammalian inferior colliculus. The large number of cell types in these midbrain sensory nuclei, compared to the number of afferent inputs (seven or more for the torus) is notable and may reflect the parcellation of function associated with the parallel processing of these inputs.

Peripheral organization and central projections of the electrosensory nerves in gymnotiform fish.

The electrosensory system of weakly electric gymnotiform fish is described from the receptor distribution on the body surface to the termination of the primary afferents in the posterior lateral line lobe (PLLL). There are two types of electroreceptor(ampullary and tuberous) and a single type of lateral line mechanoreceptor (neuromast). Receptor counts in Apteronotus albifrons show that (1) neuromasts are distributed as in other teleosts; (2) ampullary receptors number 151 on one side of the head and 208 on one side of the body; (3) tuberous receptors were estimated to number 3,000-3,500 on one side of the head and 3,500-5,000 on one side of the body. The distribution of each receptor type is described. Each receptor is innervated by a single primary afferent. Electrosensory afferents have myelinated cell bodies in the ganglion of the anterior lateral line nerve (ALLN). The distribution of these ganglion cell diameters is strongly bimodal in Apteronotus and Eigenmannia: The smaller-diameter cells may be those which innervate ampullary electroreceptors, the larger-diameter tuberous electroreceptors. Transganglionic HRP transport techniques were used to determine the first-order connections of the anterior lateral line nerve in six species of gymnotiform fish. Small branches of the ALLN were labeled so as to determine the somatotopic organization in the PLLL. The PLLL is divided into four segments from medial to lateral, termed medial, centromedial, centrolateral, and lateral segments (Heiligenberg and Dye, '81). Representations of the head are found rostrally in each zone, and the trunk is mapped caudally in each zone. Thus there are four body maps in the PLLL. The medial segment receives ampullary input (Heiligenberg and Dye, '82) and maps the dorsoventral body axis mediolaterally, as does the tuberous centrolateral segment. The tuberous centromedial and lateral segments map the dorsoventral axis lateromedially. Thus the medial and centromedial segments meet belly to belly, the centromedial and centrolateral segments meet back to back, and the centrolateral and lateral segments meet belly to belly. Adjacent electrosensory maps within the PLLL are therefore always mirror images.

Distribution of GABAergic neurons and terminals in the auditory system of the barn owl.

Antisera to GAD (glutamic acid decarboxylase) and GABA were used to determine the distribution of GABAergic cells and terminals in the brainstem and midbrain auditory nuclei of the barn owl. The owl processes time and intensity components of the auditory signal in separate pathways, and each pathway has a distinctive pattern of GAD- and GABA-like immunoreactivity. In the time pathway, all the cells of the cochlear nucleus magnocellularis and nucleus laminaris receive perisomatic GABAergic terminals, and small numbers of GABAergic neurons surround both nuclei. The ventral nucleus of the lateral lemniscus (anterior division) contains both immunoreactive terminals and some GABAergic neurons. In the intensity pathway, dense immunoreactive terminals are distributed throughout the cochlear nucleus angularis, which also contains a small number of GABAergic neurons. The superior olive contains two GABAergic cell types and immunoreactive terminals distributed throughout the neuropil. All the neurons of the nucleus of the lateral lemniscus (ventral part) appear to be GABAergic, and this nucleus also contains a moderate number of immunoreactive terminals. Immunoreactive terminals are distributed throughout the neuropil of the ventral nucleus of the lateral lemniscus (posterior division), whereas multipolar and small fusiform GABAergic neurons predominate in the dorsal regions of the nucleus. The time and intensity pathways combine in the inferior colliculus. The central nucleus of the inferior colliculus contains a larger number of fusiform and stellate GABAergic neurons and a dense plexus of immunoreactive terminals, whereas the external nucleus contains slightly fewer immunoreactive cells and terminals. The superficial nucleus contains dense, fine immunoreactive terminals and a small number of GABAergic neurons.

Calbindin-like immunoreactivity in the central auditory system of the mustached bat, Pteronotus parnelli.

With the aid of a polyclonal antibody specific for Calbindin D-28k, we studied the distribution of this calcium-binding protein in the central auditory system of the mustached bat, Pteronotus parnelli. Components of the cochlear nucleus (CN) that were calbindin-positive (cabp(+] included the root of the auditory nerve, multipolar and globular bushy cells in the anteroventral CN, multipolar and octopus cells in the posteroventral CN, and small and medium-size cells in the dorsal CN. Not stained were spherical bushy cells of the anteroventral CN and pyramidal/fusiform cells in the dorsal CN. In the superior olivary complex, labeled cells were found in the lateral and medial nuclei of the trapezoid body, the ventral and ventromedial periolivary nuclei, and the anterolateral periolivary nucleus. No cellular labeling was seen in the lateral superior olive. In the medial superior olive, only marginal cells were cabp(+). Labeled fibers could be seen surrounding the gosts of unlabeled cells in both the latter nuclei. Most cells in the intermediate nucleus and the columnar division of the ventral nucleus of the lateral lemniscus were cabp(+). However, the dorsal nucleus was cabp(-). A group of cabp(+) cells was also seen in the paralemniscal zone. The inferior colliculus had a relatively low density of cabp(+) cells. Labeled cells were more common in the caudal half of the central nucleus, and in the external nucleus and dorsal cortex. In the auditory thalamus, nearly every cell in the medial geniculate body was cabp(+), but those in the suprageniculate nucleus and in the posterior group did not stain. Small cells in the intermediate layer and giant cells in the deep layers of the superior colliculus were densely cabp(+). In the pons, cabp(+) cells and neuropil could be seen in the medial and lateral pontine nuclei (pontine gray). In conclusion, calbindin-like immunoreactivity was found in most of the brainstem auditory system, as well as in regions associated with acoustic orientation or control of vocalization. However, except for a minority of cells of the medial superior olive, it is conspicuously absent from the nuclei receiving binaural input below the level of the inferior colliculus.

Efferent projections of the posterior lateral line lobe in gymnotiform fish.

The posterior lateral line lobe (PLLL) of gymnotoid fish has efferent projections to two midbrain regions: the nucleus praeeminentialis dorsalis (n.P.d.) and the torus semicircularis dorsalis (T.Sd.). Both ipsilateral and contralateral connections are present; the n.P. d. receives nearly equal input from both sides while the T.Sd. receives a stronger contralateral input. The PLLL projection to n.P.d. merely maps medial PLLL to ventral n.P.d. and lateral PLLL to dorsal n.P.d., thus preserving the separate topography and relative orientation of the four electrosensory maps found in the PLLL. Only PLLL pyramidal cells (basilar and nonbasilar pyramids) contribute to this projection. The four PLLL electrosensory maps converge onto T.Sd. so that they map the dorsal body surface onto medial T.Sd. and the ventral body surface onto lateral T.Sd. Pyramidal cells, spherical cells, and multipolar cells contribute to this projection. A small commissural connection links homologous segments of the PLLL; these fibers arise from polymorphic cells of the PLLL.

Laminar organization of the afferent and efferent systems of the torus semicircularis of gymnotiform fish: morphological substrates for parallel processing in the electrosensory system.

The torus semicircularis of Gymnotiform fish is an enlarged laminated midbrain structure which receives lemniscal input from electrosensory, mechanoreceptive lateral line, and auditory systems. The electrosensory input in confined to the dorsal torus, while the auditory and mechanoreceptive systems project to the ventral torus. Anterograde and retrograde techniques were used were used to determine the connections of the dorsal torus in Apteronotus and Eigenmannia. The dorsal torus can be divided into nine major laminae, each of which has distinct afferent and efferent connections. The dorsal torus receives five afferent inputs: (1) A contralateral topographic input from the posterior lateral line lobe (PLLL) projects to laminae III, V, VI, VII, VIIIB, and VIIID. (2) Eurydendroid cells of the caudal lobe of the cerebellum project contralaterally to lamina VIIIB. (3) A portion of the descending nucleus of V projects to laminae VIIIA, VIIIC, and IX. (4) Lamina I is a cap of fine myelinated fibers which may originate in the torus longitudinalis. They project to laminae II and III. (5) The ipsilateral optic tectum projects to the dorsal torus. The dorsal torus projects to six major targets: (1) Laminae VII, VIII, and IX project bilaterally to a lateral region of the diencephalon above n. preglomerulosus, herein named n. electrosensorius. An area below the dorsal thalamus receives a smaller ipsilateral projection. (2) Laminae II, V, VIvn, VII, VIII, and IX project topographically to the deeper laminae of the ipsilateral optic tectum. This projection is in spatial register with the visual map in the superficial layers of the tectum. (3) Lamina VIIID projects ipsilaterally to the lateral reticular formation. (4) All laminae other than I, VI, and VIIIB project topographically to ahe ipsilateral n. praeeminentialis, which provides a powerful descending projection to the PLLL. (5) Lamina IX projects to a dorsal pretectal area. (6) The ipsilateral inferior olive receives a projection from the dorsal torus.

Localization of AMPA-selective glutamate receptors in the auditory brainstem of the barn owl.

AMPA receptor subunit-specific antibodies were used to determine if the distribution of excitatory amino acid receptors in the owl's auditory brainstem and midbrain nuclei reflected specializations for temporal processing. Each auditory nucleus displays characteristic levels of immunostaining for the AMPA receptor subunits GluR1-4, with high levels of the subtypes which exhibit rapid desensitization (GluR4 and 2/3). In the auditory brainstem, levels of GluR2/3 and GluR4 were very high in the cochlear nucleus magnocellularis and the nucleus laminaris. The different cell types of the cochlear nucleus angularis and the superior olive were characterized by heterogeneous GluR2/3 and 4 immunostaining. GluR1 levels were very low or undetectable. In the lemniscal nuclei, most neurons contained low levels of GluR1, and dense GluR2/3 and GluR4 immunoreactivity, with high levels of GluR4 in the dendrites. Levels of GluR4 were higher in the anterior portion of the ventral nucleus of the lateral lemniscus. The divisions of the inferior colliculus could be distinguished on the basis of GluR1-4 immunoreactivity, with high levels of GluR4 and moderate levels of GluR1 in the external nucleus. No major differences were observed between the pathways for encoding time and sound level cues.

Development of calretinin immunoreactivity in the brainstem auditory nuclei of the barn owl (Tyto alba).

The early development of calretinin immunoreactivity (CR-IR) was described in the auditory nuclei of the brainstem of the barn owl. CR-IR was first observed in the auditory hindbrain at embryonic day (E17) and a day later (E18) in the inferior colliculus. In each of the auditory nuclei studied, CR-IR did not develop homogeneously, but began in the regions that map high best frequencies in the adult barn owl. In the hindbrain, CR-IR was first observed in the rostromedial regions of the cochlear nucleus magnocellularis and the nucleus laminaris, and in the dorsal regions of the nucleus angularis and in the nucleus of the lateral lemniscus. In the inferior colliculus, CR-IR began in the ventral region of the central core. The edge of these gradients moved along the future tonotopic axes during the development of all nuclei studied, until adult patterns of CR-IR were achieved about a week after hatching.

A clinical trial of radioimmunotherapy with 67Cu-2IT-BAT-Lym-1 for non-Hodgkin's lymphoma.

UNLABELLED: Encouraged by the results of 131I-Lym-1 therapy trials for patients with B-cell non-Hodgkin's lymphoma (NHL), this phase I/II clinical trial of 67Cu-2IT-BAT-Lym-1 was conducted in an effort to further improve the therapeutic index of Lym-1-based radioimmunotherapy. Lym-1 is a mouse monoclonal antibody that preferentially targets malignant lymphocytes. 67Cu has beta emissions comparable to those of 131I but has gamma emissions more favorable for imaging. The macrocyclic chelating agent 1,4,7,11-tetraazacyclotetradecane-N,N',N",N"'-tetraacetic acid binds 67Cu tightly to form a stable radioimmunoconjugate in vivo. METHODS: All 12 patients had stage III or IV NHL that had not responded to standard therapy; 11 had intermediate- or high-grade NHL. At 4-wk intervals, patients received up to four doses of 67Cu-2IT-BAT-Lym-1, 0.93 or 1.85-2.22 GBq/m2 (25 or 50-60 mCi/m2), with the lower dose used when NHL was detected in the bone marrow. RESULTS: 67Cu-2IT-BAT-Lym-1 provided good imaging of NHL and favorable radiation dosimetry. The mean radiation ratios of tumor to body and tumor to marrow were 28:1 and 15:1, respectively. Tumor-to-lung, -kidney and -liver radiation dose ratios were 7.4:1, 5.3:1 and 2.6: 1, respectively. This 67Cu-2IT-BAT-Lym-1 trial for patients with chemotherapy-resistant NHL had a response rate of 58% (7/12). No significant nonhematologic toxicity was observed. Hematologic toxicity, especially thrombocytopenia, was dose limiting. CONCLUSION: 67Cu remains an option for future clinical trials. This study established 67Cu-2IT-BAT-Lym-1 as a safe, effective treatment for patients with NHL.