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.

Acoustic noise during functional magnetic resonance imaging.

Functional magnetic resonance imaging (fMRI) enables sites of brain activation to be localized in human subjects. For studies of the auditory system, acoustic noise generated during fMRI can interfere with assessments of this activation by introducing uncontrolled extraneous sounds. As a first step toward reducing the noise during fMRI, this paper describes the temporal and spectral characteristics of the noise present under typical fMRI study conditions for two imagers with different static magnetic field strengths. Peak noise levels were 123 and 138 dB re 20 microPa in a 1.5-tesla (T) and a 3-T imager, respectively. The noise spectrum (calculated over a 10-ms window coinciding with the highest-amplitude noise) showed a prominent maximum at 1 kHz for the 1.5-T imager (115 dB SPL) and at 1.4 kHz for the 3-T imager (131 dB SPL). The frequency content and timing of the most intense noise components indicated that the noise was primarily attributable to the readout gradients in the imaging pulse sequence. The noise persisted above background levels for 300-500 ms after gradient activity ceased, indicating that resonating structures in the imager or noise reverberating in the imager room were also factors. The gradient noise waveform was highly repeatable. In addition, the coolant pump for the imager's permanent magnet and the room air-handling system were sources of ongoing noise lower in both level and frequency than gradient coil noise. Knowledge of the sources and characteristics of the noise enabled the examination of general approaches to noise control that could be applied to reduce the unwanted noise during fMRI sessions.

Imaging subcortical auditory activity in humans.

There is a lack of physiological data pertaining to how listening humans process auditory information. Functional magnetic resonance imaging (fMRI) has provided some data for the auditory cortex in awake humans, but there is still a paucity of comparable data for subcortical auditory areas where the early stages of processing take place, as amply demonstrated by single-unit studies in animals. It is unclear why fMRI has been unsuccessful in imaging auditory brain-stem activity, but one problem may be cardiac-related, pulsatile brain-stem motion. To examine this, a method eliminating such motion (using cardiac gating) was applied to map sound-related activity in the auditory cortices and inferior colliculi in the brain stem. Activation in both the colliculi and cortex became more discernible when gating was used. In contrast with the cortex, the improvement in the colliculi resulted from a reduction in signal variability, rather than from an increase in percent signal change. This reduction is consistent with the hypothesis that motion or pulsatile flow is a major factor in brain-stem imaging. The way now seems clear to studying activity throughout the human auditory pathway in listening humans.

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.

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.

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.

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.

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.

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.

Comparison of cat and human brain-stem auditory evoked potentials.

Brain-stem auditory evoked potentials (BAEPs) elicited by clicks were recorded from both humans and cats. The responses of the two species were compared as functions of click level, click rate, ear stimulated, and electrode position. Since the BAEPs appear to have both high- and low-frequency components, the responses were filtered to analyze these components separately. The similarities and differences in the behavior of the peaks of the two species support the view that the first three (positive and negative) high-frequency peaks which are comparably numbered have similar generators, but the later comparably numbered peaks do not. The presence of binaural interaction beginning with P4 and PV suggests a correspondence between peaks P4 through P5 in cat with PV through PVI, respectively, in human. The similarity in behavior of these peaks also support this correspondence. Furthermore, when conduction times are estimated from interpeak latencies, this correspondence of peaks agrees more closely with the relative pathway lengths in the two species, than does the correspondence based on comparable numbering.

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.

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.

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.

Click-evoked gross potentials and single-unit thresholds in acoustically traumatized cats.

Click-evoked gross potentials were recorded from the round windows of 29 cats previously exposed to high-level sounds. The latency and amplitude of the gross neural components of these responses were determined and compared with the patterns of threshold shift measured in single auditory nerve fibers from the same 29 animals. Both of these electrophysiological measures were compared with the patterns of hair cell loss as seen in celloidin sections through the temporal bone. The correlations between single-unit abnormalities and cochlear pathology in these cases have been documented elsewhere. In this report, the correlations between gross-potential abnormalities and cochlear pathology are examined. The diagnostic potential of these correlations is discussed.

Speech coding in the auditory nerve: I. Vowel-like sounds.

Discharge patterns of auditory-nerve fibers in anesthetized cats were recorded in response to a set of nine steady-state, two-formant vowels presented at 60 and 75 dB SPL. The largest components in the discrete Fourier transforms of period histograms were almost always harmonics of the vowel fundamental frequency that were close to one of the formant frequencies, the fundamental frequency or the fiber characteristic frequency (CF). For any fiber, the position of its CF relative to the formant frequencies (F1 and F2) appears to determine which of these components dominates the response. Specifically, the response characteristics of the tonotopically arranged array of fibers can be described in terms of five CF regions: (1) a low-CF region below F1 in which the largest response components are the harmonics of the fundamental frequency closest to CF; (2) a region centered around CF = F1 in which the first formant and its harmonics are the largest components; (3) an intermediate region between F1 and F2 with prominent components at both the fiber CF and the fundamental frequency; (4) a region centered around CF = F2 in which harmonics close to the second formant are the largest for frequencies above the fundamental; and (5) a high-CF region in which response spectra tend to show broad, multiple peaks at the formant and fundamental frequencies. These CF regions are related to the phonetic descriptions of vowels. For example, the extent of the low-CF region is largest for "open" vowels (which have a high F1), and the intermediate region is distinct only for "spread" vowels for which F1 and F2 are more than 1.5-2 octaves apart. For all vowels, response activity for the majority of fibers is concentrated near the formant frequencies, in contrast to responses to broadband noise for which components near CF are dominant.

Speech coding in the auditory nerve: III. Voiceless fricative consonants.

Responses of auditory-nerve fibers in anesthetized cats were recorded for synthetic voiceless fricative consonants. The four stimuli (/x/, /s/, /s/, and /f/) were presented at two levels corresponding to speech in which the levels of the vowels would be approximately 60 and 75 dB SPL, respectively. Discharge patterns were characterized in terms of PST histograms and their power spectra. For both stimulus levels, frequency regions in which the stimuli had considerable energy corresponded well with characteristic-frequency (CF) regions in which average discharge rates were the highest. At the higher level, the profiles of discharge rate against CF were more distinctive for the stimulus onset than for the central portion. Power spectra of PST histograms had large response components near fiber characteristic frequencies for CFs up to 3-4 kHz, as well as low-frequency components for all fibers. The relative amplitudes of these components varied for the different stimuli. In general, the formant frequencies of the fricatives did not correspond with the largest response components, except for formants below about 3 kHz. Processing schemes based on fine time patterns of discharge that were effective for vowel stimuli generally failed to extract the formant frequencies of fricatives.