Efficiency of photosynthesis in a Chl d-utilizing cyanobacterium is comparable to or higher than that in Chl a-utilizing oxygenic species.

The cyanobacterium Acaryochloris marina uses chlorophyll d to carry out oxygenic photosynthesis in environments depleted in visible and enhanced in lower-energy, far-red light. However, the extent to which low photon energies limit the efficiency of oxygenic photochemistry in A. marina is not known. Here, we report the first direct measurements of the energy-storage efficiency of the photosynthetic light reactions in A. marina whole cells, and find it is comparable to or higher than that in typical, chlorophyll a-utilizing oxygenic species. This finding indicates that oxygenic photosynthesis is not fundamentally limited at the photon energies employed by A. marina, and therefore is potentially viable in even longer-wavelength light environments.

Hair-cell innervation by spiral ganglion cells in adult cats.

A horseradish peroxidase technique was used to trace the peripheral terminations of two types of ganglion cells in adult cats. It was found that large, usually bipolar ganglion cells end on inner hair cells and small, usually pseudomonopolar ganglion cells end on outer hair cells. Thus, a virtually complete segregation of afferent neural inputs from the two types of hair cells was directly confirmed.

Single unit activity in the posteroventral cochlear nucleus of the cat.

Single unit activity in the posteroventral cochlear nucleus (PVCN) was recorded for a variety of stimulus conditions. The units were classified according to their response characteristics. The locations of units were plotted onto a three-dimensional block model of the cochlear nucleus. Certain types of units that responded best to the onsets of stimuli were located predominantly in the octopus cell region of the PVCN. The remainder of the PVCN, which contains a rather heterogeneous collection of small and multipolar cells, was found to contain several types of units with the dominant type being "chopper" units.

Single unit activity in the dorsal cochlear nucleus of the cat.

Single unit activity was examined in three component layers of the dorsal cochlear nucleus (DCN): the molecular layer, the fusiform cell layer, and the polymorphic layer (deep DCN). Electrophysiological units were classified into types on the basis of their activity under a variety of stimulus conditions. In the molecular layer spike activity was small and difficult to isolate. Almost all units in the fusiform cell layer could be classified as either "pauser" or "buildup" units. Classification of units in the deep DCN was sometimes difficult, but "pauser," "chopper," and some "on" units were found. The "on" types of units tended to be located in the more superficial part of the deep DCN. Unit locations were referred to a three-dimensional block model of the cochlear nucleus.

Unmyelinated axons of the auditory nerve in cats.

This paper describes some central terminations of type II spiral ganglion neurons as labeled by extracellular injections of horseradish peroxidase (HRP) into the auditory nerve of cats. After histological processing with diaminobenzidine, both thick (2-4 microns) and thin (0.5 microns) fibers of the auditory nerve were stained. Whenever traced, thick fibers always originated from type I spiral ganglion neurons and thin fibers always from type II ganglion neurons. Because the labeling of type II axons faded as fibers projected into the cochlear nucleus, this report is limited to regions of the ventral cochlear nucleus near the auditory nerve root. The central axons of type II neurons are unmyelinated, have simple yet variable branching patterns in the cochlear nucleus, and form both en passant and terminal swellings. Under the light microscope, most swellings are located in the neuropil but they are also found in the vicinity of cell bodies, nodes of Ranvier of type I axons, and blood vessels. Eighteen en passant swellings in the neuropil were located by light microscopy and resectioned for electron microscopy; two of these swellings exhibited ultrastructural features characteristic of chemical synapses. The data indicate that inputs from outer hair cells might be able to influence auditory processing in the cochlear nucleus through type II primary neurons.

Central trajectories of type II spiral ganglion neurons.

Previous attempts to trace the central pathways of the thin axons from type II spiral ganglion neurons have been hampered by technical difficulties such as fading of the reaction product as distance increases from the injection site (Ryugo et al.: Soc. Neurosci. Abstr. 12:779, '86; Brown: J. Comp. Neurol. 260:591-604, '87). By using small rodents (gerbils and mice), which have short auditory nerves, we have succeeded in filling the entire central axon and terminals of type II neurons after peripheral injections of horseradish peroxidase. The general course of the type II fibers within the auditory nerve and cochlear nucleus is similar to that of type I fibers except that terminals from type II neurons are often found in regions of the cochlear nucleus that have high densities of granule cells.

Number and distribution of stapedius motoneurons in cats.

Cell bodies of stapedius motoneurons were identified by retrograde transport of horseradish peroxidase (HRP) following injections into the stapedius muscle. Large injections were made in an attempt to label all stapedius motoneurons. To control for labeling of non-stapedial neurons resulting from spread of HRP, we determined the locations of brainstem neurons labeled by HRP applied to the facial nerve, the chorda tympani nerve, the auricular branch of the vagus nerve, the tensor tympani muscle, and the cochlea. In three cats analyzed in detail, 1,133-1,178 neurons projecting to the stapedius muscle were identified. Arguments are given which suggest that in these three cats all stapedius motoneurons were labeled. The labeled stapedius neurons may all be motoneurons because they all stain positively for acetylcholinesterase and have medium-coarse Nissl bodies. Most stapedius motoneurons were located around the motor nucleus of the facial nerve. Staphedius motoneurons were also found near the descending limb of the facial-nerve root, in the peri-olivary neuropil, and in the reticular formation with the ascending fibers of the facial-nerve root.

A block model of the cat cochlear nucleus.

A three-dimensional block model of the cochlear nucleus of the cat was constructed from histologic sections. Boundaries of various subdivisions, based on cytoarchitectonic criteria, were included in the model. Usage of the block model in correlating physiological and anatomical data is illustrated by localizing characteristic waveforms of gross evoked responses and characteristic frequencies of single units.

Curious oddments of auditory-nerve studies.

Three interesting theoretical issues are presented to illustrate how certain isolated observations on auditory-nerve activity can be puzzling until other, seemingly unrelated phenomena are documented. The issues are (1) disinhibition; (2) 'peak-splitting'; and (3) independence of spike generation in primary neurons innervating the same inner-hair cell. (1) The issue of disinhibition is important for theories of lateral inhibition. For auditory-nerve fibers, the question can he phrased, 'If the rate of discharge to a tone at the characteristic frequency (CF) of a unit can he reduced by adding a second tone off the CF, is it possible to suppress this reduction by adding a third tone, even further off the CF?' The data are insufficient to conclude that disinhibition is found for auditory-nerve fibers and other explanations are available to account for the results of three-tone experiments. (2) Normally, only a single peak in the histogram of responses to low tones is phase-locked, but at high stimulus levels, the histograms will show two, or even three, peaks per stimulus cycle ('peak-splitting'). At still higher levels, the histograms again show only a single peak, but it is phase-shifted from the original peak for low stimulus levels. This complex sequence of events can be accounted for by simple models. (3) Although simultaneous recordings from pairs of auditory-nerve fibers have failed to show non-stimulus related correlations between spike trains, it has not been directly demonstrated that any two recorded fibers innervate the same hair cell. However, an indirect argument is offered to support the idea that fibers innervating a single inner-hair cell must have independent spike generators.

The effect of brainstem lesions on brainstem auditory evoked potentials in the cat.

Brainstem auditory evoked potentials (BAEPs) were recorded before and after cuts were made in either the midline trapezoid body (TB), the lateral lemniscus (LL), or the combined dorsal and intermediate acoustic striae (DAS/IAS) in 23 anesthetized cats. Monaural and binaural rarefaction clicks were presented at a rate of 10 per s, and the potentials recorded from a vertex electrode referenced to either earbar or to the neck. The potentials were filtered so that fast and slow components could be examined separately and special efforts were exerted to obtain stable conditions so that small changes in waveforms could be significant. Lesions of the DAS/IAS produced negligible changes in either the fast or slow waves. Lesions of the midline TB reduced the amplitudes of peaks P3 through P5, while greatly reducing the amplitude of the slow wave. Complete lesions of the LL always reduced the amplitude of the slow wave. Lesions of the ventral part of the LL were more likely to reduce the amplitude of P4-P5. Our interpretations of these lesion experiments are based on the idea that individual fast peaks of the BAEP represent compound action potentials of fiber pathways. According to this view, only synchronized activity generated in populations of neurons that are both favorably oriented in space and significant in number, will contribute to the fast peak.

Generators of the brainstem auditory evoked potential in cat. III: Identified cell populations.

This paper examines the relationship between different brainstem cell populations and the brainstem auditory evoked potential (BAEP). First, we present a mathematical model relating the BAEP to underlying cellular activity. Then, we identify specific cellular generators of the click-evoked BAEP in cats by combining model-derived insights with key experimental data. These data include (a) a correspondence between particular brainstem regions and specific extrema in the BAEP waveform, determined from lesion experiments, and (b) values for model parameters derived from published physiological and anatomical information. Ultimately, we conclude (with varying degrees of confidence) that: (1) the earliest extrema in the BAEP are generated by spiral ganglion cells, (2) P2 is mainly generated by cochlear nucleus (CN) globular cells, (3) P3 is partly generated by CN spherical cells and partly by cells receiving inputs from globular cells, (4) P4 is predominantly generated by medial superior olive (MSO) principal cells, which are driven by spherical cells, (5) the generators of P5 are driven by MSO principal cells, and (6) the BAEP, as a whole, is generated mainly by cells with characteristic frequencies above 2 kHz. Thus, the BAEP in cats mainly reflects cellular activity in two parallel pathways, one originating with globular cells and the other with spherical cells. Since the globular cell pathway is poorly represented in humans, we suggest that the human BAEP is largely generated by brainstem cells in the spherical cell pathway. Given our conclusions, it should now be possible to relate activity in specific cell populations to psychophysical performance since the BAEP can be recorded in behaving humans and animals.

Single-neuron labeling and chronic cochlear pathology. IV. Stereocilia damage and alterations in rate- and phase-level functions.

The rate and phase of auditory-nerve response to tone bursts were studied as a function of stimulus level in normal and acoustically traumatized animals. The rate- and phase-level functions of normal auditory-nerve fibers are often separable into a low-intensity component (component I) and high-intensity component (component II), as defined by a dip in the rate function and a simultaneous abrupt shift in the phase function at stimulus levels near 90 dB SPL [10,12,9]. Baseline data are established by defining the relation between stimulus frequency and the characteristic frequency and spontaneous discharge rate of a fiber normally required for the appearance of these two components in the response. Abnormalities of the level functions are shown to occur in acoustically traumatized ears. Noise-induced threshold shift is often characterized by selective attenuation of component I. In some instances, it appears that component I has been eliminated, leaving a response which is identical in threshold, phase and maximum discharge rate to a normal component II. Results of single-unit labeling in such a case suggest that the selective attenuation of component I is associated with selective loss of the tallest row of stereocilia on the inner hair cells (IHCs). It is suggested that component I is normally generated through an interaction between the outer hair cells and the tall row of IHC stereocilia, while component II requires only the shorter row of IHC stereocilia.

Single unit clues to cochlear mechanisms.

In recent years studies on isolated hair cells have suggested that there is an inherent tuning of hair cells determined by their mechanical and electrical properties. However, tuning for mammalian cochleas appears to be much more complicated since there are typically two types of receptor cells (inner and outer hair cells) imbedded in a highly organized framework of supporting cells, membranes and fluids. The major neural output of the cochlea can be monitored by recording the activity of myelinated axons of spiral ganglion cells, not only under normal conditions, but also when the discharge patterns are altered by ototoxic drugs, acoustic trauma or olivocochlear bundle stimulation. A model system with two excitatory influences, one sharply tuned and highly sensitive, and a second, broadly tuned and relatively insensitive, can account for much of the existing data. Results from single-neuron marking studies support the notion that these two influences probably involve interactions between inner and outer hair cells. More global influences such as the endocochlear potential also can act on auditory-nerve fibers through the hair-cell systems. Thus, the inherent frequency selectivity of the receptor cell is only one of many factors that determine the tuning of mammalian auditory-nerve fibers.

Generators of the brainstem auditory evoked potential in cat. II. Correlating lesion sites with waveform changes.

Brainstem regions involved in generating the brainstem auditory evoked potential (BAEP) were identified by examining the effects of lesions on the click-evoked BAEP in cats. An excitotoxin, kainic acid, was injected into various parts of the cochlear nucleus (CN) or into the superior olivary complex (SOC). The locations of the resulting lesions were correlated with the changes produced in the various extrema of the BAEP waveforms. The results indicate that: (1) the earliest BAEP extrema (P1, N1 (recorded between vertex and the earbar ipsilateral to the stimulus) and P1a, P1b, (vertex to contralateral earbar)) are generated by cells with somata peripheral to the CN; (2) P2 is primarily generated by posterior anteroventral CN (AVCNp) and anterior posteroventral CN (PVCNa) cells; (3) SOC, anterior anteroventral CN (AVCNa), AVCNp, and PVCNa cells are involved in generating P3; (4) AVCNa cells are the main CN cells involved in P4, N4, and P5 generation; (5) both ipsilateral and contralateral SOC cells have a role in generating monaurally evoked P4 and P5; and (6) P5 is generated by cells with characteristic frequencies below 10 kHz. From (2) and (4), it is clear that P2 and P4-P5 are generated by cells in distinct, parallel pathways.

Binaural auditory processing in multiple sclerosis subjects.

In order to relate human auditory processing to physiological and anatomical experimental animal data, we have examined the interrelationships between behavioral, electrophysiological and anatomical data obtained from human subjects with focal brainstem lesions. Thirty-eight subjects with multiple sclerosis were studied with tests of interaural time and level discrimination (just noticeable differences or jnds), brainstem auditory evoked potentials and magnetic resonance (MR) imaging. Interaural testing used two types of stimuli, high-pass (> 4000 Hz) and low-pass (< 1000 Hz) noise bursts. Abnormal time jnds (Tjnd) were far more common than abnormal level jnds (70% vs 11%); especially for the high-pass (Hp) noise (70% abnormal vs 40% abnormal for low-pass (Lp) noise). The HpTjnd could be abnormal with no other abnormalities; however, whenever the BAEPs, LpTjnd and/or level jnds were abnormal HpTjnd was always abnormal. Abnormal wave III amplitude was associated with abnormalities in both time jnds, but abnormal wave III latency with only abnormal HpTjnds. Abnormal wave V amplitude, when unilateral, was associated with a major HpTjnd abnormality, and, when bilateral, with both HpTjnd and LpTjnd major abnormalities. Sixteen of the subjects had their MR scans obtained with a uniform protocol and could be analyzed with objective criteria. In all four subjects with lesions involving the pontine auditory pathway, the BAEPs and both time jnds were abnormal. Of the twelve subjects with no lesions involving the pontine auditory pathway, all had normal BAEPs and level jnds, ten had normal LpTjnds, but only five had normal HpTjnds. We conclude that interaural time discrimination is closely related to the BAEPs and is dependent upon the stimulus spectrum. Redundant encoding of low-frequency sounds in the discharge patterns of auditory neurons, may explain why the HpTjnd is a better indicator of neural desynchrony than the LpTjnd. Encroachment of MS lesions upon the pontine auditory pathway always is associated with abnormal BAEPs and abnormal interaural time discrimination but may have normal interaural level discrimination. Our data provide one of the most direct demonstrations in humans of relationships among auditory performance, evoked potentials and anatomy. We present a model showing that many of these interrelationships can be readily interpreted using ideas developed from work on animals, even though these relationships could not have been predicted with confidence beforehand. This work provides a clear advance in our understanding of human auditory processing and should serve as a basis for future studies.

Effects of multiple sclerosis brainstem lesions on sound lateralization and brainstem auditory evoked potentials.

Magnetic resonance (MR) imaging, brainstem auditory evoked potentials (BAEPs), and tests of interaural time and level discrimination were performed on sixteen subjects with multiple sclerosis (MS). Objective criteria were used to define MR lesions. Of the eleven subjects in whom no pontine lesions were detected and the one subject who had pontine lesions that did not encroach upon the auditory pathways, all had normal BAEPs and interaural level discrimination, although a few had abnormal interaural time discrimination. Of four subjects with lesions involving the pontine auditory pathway, all had both abnormal BAEPs and abnormal interaural time discrimination; one also had abnormal interaural level discrimination. Analysis of the data suggest the following: waves I and II are generated peripheral to the middle of the ventral acoustic stria (VAS); wave III is generated ipsilaterally in the region of the rostral VAS, caudal superior olivary complex (SOC) and trapezoid body (TB); and waves V and L are generated contralaterally, rostral to the SOC-TB. The region of the ipsilateral rostral SOC-TB is implicated as part of the pathway involved in the generation of waves V and L. Interaural time discrimination of both high and low frequency stimuli were affected by all brainstem lesions that encroached on auditory pathways. A unilateral lesion in the region of the LL affected interaural time discrimination for low-frequency stimuli less severely than bilateral lesions of the LL or a unilateral lesion of the VAS. The only interaural level discrimination abnormality occurred for a subject with a unilateral lesion involving the entire rostral VAS. It appears that detailed analysis of lesion locations coupled with electrophysiological and psychophysical data holds promise for testing hypotheses concerning the function of various human auditory brainstem structures.

Generators of the brainstem auditory evoked potential in cat. I. An experimental approach to their identification.

This paper is the first in a series aimed at identifying the cellular generators of the brainstem auditory evoked potential (BAEP) in cats. The approach involves (1) developing experimental procedures for making small selective lesions and determining the corresponding changes in BAEP waveforms, (2) identifying brainstem regions involved in BAEP generation by examining the effects of lesions on the BAEP and (3) identifying specific cell populations involved by combining the lesion results with electrophysiological and anatomical information from other kinds of studies. We created lesions in the lower brainstem by injecting kainic acid which is generally toxic for neuronal cell bodies but not for axons and terminals. This first paper describes the justifications for using kainic acid, explains the associated problems, and develops a methodology that addresses the main difficulties. The issues and aspects of the specific methods are generally applicable to physiological and anatomical studies using any neurotoxin, as well as to the present BAEP study. The methods chosen involved (1) measuring the BAEP at regular intervals until it reached a post-injection steady state and perfusing the animals with fixative shortly after the last BAEP recordings were made, (2) using objective criteria to distinguish injection-related BAEP changes from unrelated ones, (3) making control injections to identify effects not due to kainic acid toxicity, (4) verifying the anatomical and functional integrity of axons in lesioned regions, and (5) examining injected brainstems microscopically for cell loss and cellular abnormalities indicating dysfunction. This combination of methods enabled us to identify BAEP changes which are clearly correlated with lesion locations.

Auditory-nerve activity in cats exposed to ototoxic drugs and high-intensity sounds.

The response characteristics of auditory-nerve fibers in normal cats are compared with those in cats exposed to kanamycin and high-intensity sounds. The pathophysiology is characterized by an elevation of the tuning-curve "tips," which is sometimes associated with hypersensitivity of the "tails". Plots of unit thresholds are correlated with patterns of sensory-cell losses in the cochlea. There can be significant shifts in unit threshold without significant loss of hair cells; however, significant hair cell loss is always accompanied by highly abnormal unit thresholds. The presence of inner hair cells seems to be essential for the long-term survival of spiral ganglion cells. An incidental observation is that in the "normal" animal there is almost always a prominent "notch" at 3-4 kHz in the plots of threshold at characteristic frequency, which may have been produced by environmental noise.

Recording auditory-nerve potentials as an office procedure.

Recording auditory-nerve potentials from human subjects is already a routine procedure in the laboratory. In order to bring such recording capabilities into the office of practicing otologists, a number of difficulties had to be overcome. First, a small signal averager was built and incorporated into a stimulus generating and response recording system. The entire system was made portable and self-sufficient. The effects of electrical interference and background acoustic noise were shown to be tolerable. After studies of how responses vary with electrode location, electrodes were designed to be placed on the ear canal so that no invasive procedures were necessary. Methods were found to simplify the procedure so that recordings can be made in a matter of minutes by one person working alone.

Fundamental considerations in designing auditory implants.

The usual premise underlying developmental work on cochlear prostheses as prospective cures for profound deafness is that the auditory nerve can be electrically stimulated in such a manner that communicative skills can be developed or maintained. Physiologic recordings from single fibers in the cat's auditory nerve and attempts to model these responses have generated a description of how the auditory nerve codes complex sounds such as speech. This work suggests that certain minimal cues might have to be present at the level of the auditory nerve in order that adequate discrimination of specific speech signals can take place. The prospects for achieving a useful prosthesis in the near future will be evaluated in terms of what can be expected from current attempts to code the artificial stimulation properly.