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Biomedical subjects

C P Richter

Publications and source records attributed to C P Richter.

At least 19 recordsLinked to original sources
PubMed2006

Survival and morphology of auditory neurons in dissociated cultures of newborn mouse spiral ganglion.

We have systematically characterized neuronal survival and growth in cultures derived from newborn/postnatal day 1 mouse cochlea. Dissociated cultures of the cochlear spiral ganglion provide an experimental environment in which to examine molecular mechanisms of survival, development and physiology of auditory neurons. To relate survival to the total number of neurons present in the source tissue, three cochleas from different newborn CD-1 mice were embedded in Araldite resin and serially sectioned at 5 mum thickness. All neurons were counted. To avoid overcounting, each section served as a lookup section for the next, giving 8240+/-423 (S.D.) neurons per ganglion. Cultures maintained in the presence of adjacent non-neural tissue, brain-derived neurotrophic factor, neurotrophin 3, leukemia inhibitory factor (LIF) and 10% fetal bovine serum returned the best overall survival (30%) at 42 h post-plating. Best overall survival required the continuous presence of a serum component(s) larger than 100,000 MW. Plating efficiency (number of neurons that attach to the well after 4 h) was similar in the presence or absence of LIF. Inclusion of LIF maintained 100% survival of plated neurons over 42 h of culture; without LIF, a large fraction of the neurons did not survive. LIF appeared to maintain survival by preferentially preserving a population of bipolar neurons, while having little effect on the number of monopolar neurons. This work provides quantitative measures of survival and morphology of auditory neurons in vitro. The results support the idea that survival of spiral ganglion neurons in vivo may depend on interactions with adjacent, non-neural tissue and raise the possibility that maintenance of bipolar morphology after hair cell damage may require biochemical mechanisms in addition to those induced by neurotrophins.

Animals↗
PubMed2001

A reconsideration of sound calibration in the mouse.

Although it is traditional to perform sound calibrations in anesthetized animals by placing a probe-tube microphone near the tympanic membrane, these measurements are inaccurate at high stimulus frequencies where hearing must be quantified in the mouse. Hence, our motivation to develop another approach using the mouse ear canal as a coupler. Results of real-ear-canal calibrations indicate that an average calibration can be used to estimate sound pressure levels in the three mouse strains tested. Similar estimates were also obtained using a tubing coupler, whose volume was comparable to that of the ear canal, thereby offering a simpler alternative. In addition, real-head calibrations were also performed to provide a procedure that can be used in situations where the ear is not dissected, as in measurements of the auditory brainstem response. Calibrations for open, rather than closed, sound-delivery systems were also evaluated using a modified method of substitution.

Acoustics↗
PubMed2001

Cochlear dimensions obtained in hemicochleae of four different strains of mice: CBA/CaJ, 129/CD1, 129/SvEv and C57BL/6J.

Because homologies between mice and human genomes are well established and hereditary abnormalities are similar in both, mice present a valuable animal model to study hereditary hearing disorders in humans. One of the manifestations of hereditary hearing disorders might be in the structure of cochlear elements, such as the gross morphology of the cochlea. Cochlear dimensions, however, are one factor that determines inner ear mechanics and thus hearing function. Therefore, gross cochlear dimension might be important when different strains of mice are compared regarding their hearing. Although several studies have examined mouse inner ear structures on a sub-cellular level, only few have studied cochlear gross morphology. Moreover, the sparse data available were acquired from fixed and dehydrated tissue. Dehydration, however, produces severe distortion of gel-like cochlear structures such as the tectorial membrane and the basilar membrane hyaline matrix. In this study, the hemicochlea technique, which allows fresh mouse cochlear material to be viewed from a radial perspective, was used to provide an itemized study of the dimensions of gross cochlear structures in four mouse strains (CBA/CaJ, 129/SvEv, 129/CD1 and C57BL/6J). Except for the CBA/CaJ, these strains are known to possess genes for age-related hearing loss. The measurements showed no major differences among the four strains. However, when compared with previous data, the thickness measures of the basilar membrane were up to 10 times larger. Such differences are likely to result from the different techniques used to process the material. The hemicochlea technique eliminates much of the distortion caused by dehydration, which was present in previous experiments.

Animals↗
PubMed2000

Development of the gerbil inner ear observed in the hemicochlea.

A frequency-dependent change in hearing sensitivity occurs during maturation in the basal gerbil cochlea. This change takes place during the first week after the onset of hearing. It has been argued that the mass of a given cochlear segment decreases during development and thus increases the best frequency. Changes in mass during cochlear maturation have been estimated previously by measuring the changes in cochlear dimensions. Fixed, dehydrated, embedded, or sputter-coated tissues were used in such work. However, dehydration of the tissue, a part of most histological techniques, results in severe distortion of some aspects of cochlear morphology. The present experiments, using a novel preparation, the hemicochlea, show that hydrated structures, such as the tectorial membrane and the basilar membrane hyaline matrix, are up to 100% larger than estimated previous studies. Therefore, the hemicochlea was used to study the development of cochlear morphology in the gerbil between the day of birth and postnatal day 19. We used no protocols that would have resulted in severe distortion of cochlear elements. Consequently, a detailed study of cochlear morphology yields several measures that differ from previously published data. Our experiments confirm growth patterns of the cochlea that include a period of remarkably rapid change between postnatal day 6 and 8. The accelerated growth starts in the middle of the cochlea and progresses toward the base and the apex. In particular, the increase in height of Deiters' cells dominated the change, "pushing" the tectorial membrane toward scala vestibuli. This resulted in a shape change of the tectorial membrane and the organ of Corti. The tectorial membrane was properly extended above the outer hair cells by postnatal day 12. This time coincides with the onset of hearing. The basilar membrane hyaline matrix increased in thickness, whereas the multilayered tympanic cover layer cells decreased to a single band of cells by postnatal day 19. Before and after the period of rapid growth, the observed gross morphological changes are rather small. It is unlikely that dimensional changes of cochlear structures between postnatal days 12 and 19 contribute significantly in the remapping of the frequency-place code in the base of the cochlea. Instead, structural changes affecting the stiffness of the cochlear partition might be responsible for the shift in best frequency.

Animals↗
PubMed1999

Sodium, potassium, chloride and calcium concentrations measured in pigeon perilymph and endolymph.

According to Davis' (1965) model of the inner ear, a potential difference between the endocochlear potential and the hair cell resting potential drives the transduction current across the apical hair cell membrane. It is assumed that the endocochlear potential (EP) consists of two components. The first is a diffusion potential, which depends on the ionic composition of endolymph and perilymph and on the permeability of the perilymph-endolymph barrier. The second is an electrogenic component which is determined by active ion transport across the perilymph-endolymph barrier. In birds, the EP is between +8 and +20 mV. Little is known about the underlying mechanisms responsible for the measured EP in birds. The present paper studies whether ionic compositions of endo- and perilymph might explain the EP in birds. Concentrations of Na+, K+, Ca2+ and Cl- in pigeon scala vestibuli, scala tympani and scala media were determined with ion-selective microelectrodes. Na+, K+, Ca2+ and Cl- were 150.0, 4.2, 1.4 and 117.0 mM in perilymph (scala tympani and scala vestibuli). In scala media, the concentrations of K+, Ca2+ and Cl- were 140.6, 0.23 and 142.1 mM.

Animals↗
PubMed1999

Evidence for an anion exchange mechanism for uptake of conjugated bile acid from the rat jejunum.

Absorption of conjugated bile acids from the small intestine is very efficient. The mechanisms of jejunal absorption are not very well understood. The aim of this study was to clarify the mechanism of absorption of conjugated bile acid at the apical membrane of jejunal epithelial cells. Brush-border membrane vesicles from intestinal epithelial cells of the rat were prepared. Absorption of two taurine-conjugated bile acids that are representative of endogenous bile acids in many variate vertebrate species were studied. In ileal, but not jejunal brush-border membrane vesicles, transport of conjugated bile acids was cis-stimulated by sodium. Transport of conjugated bile acids was trans-stimulated by bicarbonate in the jejunum. Absorption of conjugated dihydroxy-bile acids was almost twice as fast as of trihydroxy-bile acids. Coincubation with other conjugated bile acids, bromosulfophthalein, and DIDS, as well as by incubation in the cold inhibited the transport rate effectively. Absorption of conjugated bile acids in the jejunum from the rat is driven by anion exchange and is most likely an antiport transport.

Absorption↗
PubMed1998

Morphology of the unfixed cochlea.

Our knowledge of cochlear geometry is based largely upon anatomical observations derived from fixed, dehydrated, embedded and/or sputter-coated material. We have now developed a novel preparation, the hemicochlea, where for the first time living cochlear structures can be observed in situ and from a radial perspective. The experiments were performed on the Mongolian gerbil. Ion substitution experiments suggest that no significant swelling or shrinkage occurs when the preparation is bathed in normal culture medium, so long as calcium concentration is kept at endolymph-like (20 microM) levels. The tectorial membrane-reticular lamina relationship appears to remain well preserved. Hensen's stripe maintains a close relationship with the inner hair cell stereociliary bundle, unless the mechanical coupling becomes disturbed. In addition, standard fixation and/or dehydration procedures are used to quantify changes due to shrinkage artifacts. Various morphometric gradients are examined in unfixed specimens from apical, middle, and basal turns.

Animals↗
PubMed1998

Basilar membrane vibration in the gerbil hemicochlea.

Excised gerbil cochleae were cut along the mid-modiolar plane (hemicochlea). Along one-half turn of this preparation, fluorescent microbeads were placed on the basilar membrane (BM). The BM was vibrated with click stimuli (50 micros) produced mechanically by a piezo pusher. The stimulus delivery probe could be positioned either more apical or more basal from the beads. Vibration patterns were measured with a wide bandwidth photomultiplier from the movements of the beads. When the probe was positioned more basal, the responses to click stimuli were brief, damped sinusoids. According to the fast Fourier transforms (FFTs) of the averaged time wave forms, the best frequency between successive beads decreased toward the apex (0.8 octave/mm). Sharpness of tuning of the normalized FFT spectra (NQ10dB) on average was 1.5. Response amplitude at a fixed input level, measured at different beads away from the stimulation site, dropped exponentially (58 dB/mm). In addition, for each individual bead, amplitude dropped linearly with decreasing stimulus intensity. In experiments where the stimulating probe was placed more apical, two major properties were observed: first, beads revealed only the spectral components present in the motion of the probe. Second, magnitude reduction of the displacement of the cochlear partition was greater, on average 155 dB/mm, indicating a lack of significant propagation in the reverse direction.

Acoustic Stimulation↗
PubMed1997

Modeling stochastic spike train responses of neurons: an extended Wiener series analysis of pigeon auditory nerve fibers.

A general method for developing data-based, stochastic nonlinear models of neurons by means of extended functional series expansions was applied to neural activities of pigeon auditory nerve fibers responding to Gaussaian white noise stimuli. To determine Wiener series representations of the investigated neurons the fast orthogonal search algorithm was used. The results suggest that nonlinearities are only instantaneous and that the signal transduction of the investigated sensory system can be described by cascades of dynamic linear and static nonlinear devices. However, only slight improvements result from the nonlinear terms. Considerable improvements are, nevertheless, possible by generalizing the ordinary Wiener series, so that prior neural activity can be taken into account. These extended series were used to develop stochastic models of spiking neurons. The models are able to generate realistic interspike interval distributions and rate-intensity functions. Finally, it will be shown that the irregularity in real and modeled action potential trains has advantages concerning the decoding of neural responses.

Action Potentials↗
PubMed1996

Development of activity patterns in auditory nerve fibres of pigeons.

Little is known about inner ear development in pigeons. This paper addresses the question of maturation in activity patterns of pigeon auditory nerve fibres, Pigeons that were 1, 2 and 4 weeks and 1, 2, 3 and 4 years old were investigated. Adult-like activity patterns are found 4 weeks post-hatching. Spontaneous activities of fibres in immature animals (about 40 spikes/s) are half that found in adults. Spontaneous discharge rate does not increase with decreasing characteristic frequency (CF) of the fibre if the animals are immature. Rate threshold are less sensitive in immature animals. Differences between the age groups are generally significant if the CFs of the fibres are below 1.3 kHz. Sharpness of tuning is already adult-like in l-week-old animals. Inter-spike time interval histograms (ISTH) of auditory fibres recorded in animals of all age groups often show Poisson-like distributions. Preferred intervals are found in 10% of the ISTHs of fibres in immature animals but in 30% of adults. Cross-correlations between heart beats of the animal and spontaneous activities show good correlation for about 70% of the fibres in immature animals. With the growth of the animals, the number of fibres showing correlation of spontaneous activities and heart beats decreases to about 40%. The basilar papilla of a 1-week-old animal is smaller than in an adult animal (by 10% in length and by 10% in width), judge by scanning electron microscopy (SEM), Changes of activity patterns in this study are likely to be a result of maturation of the middle ear. In addition to the latter, development of the inner ear is conceivable.

Aging↗
PubMed1995

Rate-intensity-functions of pigeon auditory primary afferents.

Rate-intensity-functions (RI-functions) were determined in 150 primary auditory afferents in anaesthetized pigeon. Acoustic stimulation was either at characteristic frequency (CF) or half an octave below or above CF. Stimulated at CF, 37% of the fibres showed saturating RI-functions, whereas 50% showed sloping and 13% straight RI-functions. In the sloping RI-functions, a bend was found about 20 dB above the fibres' thresholds. For non-CF stimuli, the general shape of the RI-functions remained constant. However, the maximum evoked discharge rates were lower for frequencies below CF and higher for frequencies above CF. The data show that a population of neurones, the sloping and straight ones, code stimulus intensities over a wide intensity range. In combination with the scatter of the thresholds, intensity ranges greater than 100 dB are conceivable. It was concluded that the nonlinearities found in pigeon are not caused by basilar membrane (BM) mechanics, rather an origin at the hair cell-afferent nerve fibre system has to be considered.

Acoustic Stimulation↗
PubMed1994

Artifact-free application of drugs into the avian endolymphatic space.

A push-pull system is described that allows artifact-free application of drugs into very small volumes such as, e.g., the endolymphatic space of the inner ear. The system consists of two syringes, the pistons of which are fixed together and driven by an Oriel motordrive. The syringes are connected to the pipette holders and microcapillaries by PVC tubes. One capillary is used to expel a certain volume, whilst the other one aspirates the same volume. A balanced exchange of fluids within a given space without an overall change in volume is therefore possible.

Action Potentials↗
PubMed1994

Effects of endolymphatic and perilymphatic application of salicylate in the pigeon. I: Single fiber activity and cochlear potentials.

The effects of salicylate on the mammalian cochlea function are well documented. However, there is a lack of reports on salicylate effects on the avian auditory periphery and it might well be that salicylate is not ototoxic at all in submammalian vertebrates. We therefore recorded single fiber activities, compound action potential (CAP) and endocochlear potential (EP) during application of salicylate (calculated final concentration of about 2-18 mmol/l) into the scala media of pigeons. We furthermore recorded CAP and EP during perilymphatic perfusion of salicylate (2-20 mmol/l). Salicylate applied into the scala media led to an elevation of tip threshold in single fibers ranging from 5 to 35 dB. The characteristic frequencies of the fibers were not changed. This effect on auditory nerve fibers was reflected in an elevation of CAP thresholds. The mean spontaneous discharge rate was either slightly increased or remained unchanged in the majority of fibers. Perilymphatic salicylate perfusion also led to an elevation of CAP thresholds that was reversible following subsequent perfusion with artificial perilymph. The EP remained unchanged in both application modes. The effects of salicylate were dose dependent and more pronounced in the mid- to high-frequency range. These results are consistent with an action of salicylate on the process (electrical or mechanical, or both) responsible for the sensitivity and frequency selectivity in the avian peripheral hearing organ.

Action Potentials↗
PubMed1994

Effects of endolymphatic and perilymphatic application of salicylate in the pigeon. II: Fine structure of auditory hair cells.

Large doses of salicylate are known to cause reversible ototoxic effects including fine structural alterations of the auditory hair cells in mammals. To investigate possible fine structural correlates of salicylate effects on pigeon auditory hair cells, the basilar papillae following perilymphatic or endolymphatic application of salicylate were fixed and processed for transmission electron microscopy. The pigeon auditory hair cells possessed organelles typically described in avians. A single or multi-layered array of cisternae along the cytoplasmic side of the lateral plasma membrane, i.e. subsurface cisternae that are characteristic for mammalian outer hair cells, was not seen. The most prominent fine structural alterations of hair cells after salicylate application were an increase in the luminal width of smooth and rough endoplasmic reticulum as well as the frequent occurrence of prominent single-membrane-bound vesicles filled with electron-dense bodies. Based on the assumption that subsurface cisternae represent a specialized form of endoplasmic reticulum, the present findings indicate that the structural correlates of salicylate toxicity are similar in mammalian and avian auditory hair cells.

Animals↗
PubMed1994

Preferred intervals in birds and mammals: a filter response to noise?

Quasi-periodic spontaneous activity (preferred intervals, PIs) has been reported from avian primary auditory afferents. In mammals, PIs have not been reported, as yet. As the length of PIs is close to 1/characteristic frequency, it has been suggested that this type of spontaneous activity indicates particular mechanisms in avian inner ear transduction. However, the present paper shows that pigeon auditory fibres possessing preferred intervals in their spontaneous activity always belong to the most sensitive and the most sharply-tuned fibres recorded. This leads to the assumption that preferred intervals are the response of narrow-band filters to noise. This view is supported by three additional findings: (i) Near-threshold noise provokes PIs in avian fibres that show no spontaneous PIs. (ii) Similarly, PIs can also be evoked in mammalian (gerbil) auditory afferents by low level noise. (iii) Phase-locking of auditory afferents can be achieved by sound stimuli 10-20 dB below rate threshold. It is argued that no conclusions may be drawn from the presence of PIs about the nature of the underlying filter.

Acoustic Stimulation↗
PubMed1978

"Dark-active" rat transformed into "light-active" rat by destruction of 24-hr clock: function of 24-hr clock and synchronizers.

In alternating 12-hr periods of light and dark the rat is active mainly in the dark. Its activity in the dark (beginning at 1800) depends exclusively on release of activity by the 24-hr clock. In the light (beginning at 0600) the 24-hr clock inhibits activity; the normal rat becomes totally inactive in the light except for activity resulting from external stimulation. After section of the connections between the optic chiasma and the hypothalamus, some rats become totally and permanently inactive in the dark. This sectioning destroys the 24-hr clock. After destruction of the clock removes inhibition of activity in the light period, the rat becomes active promptly at start of the light period--i.e., becomes a "light-active" animal. In the normal rat, activity becomes synchronized to start of the dark (by the electric clock at 1800), regardless of the amounts of activity. Destruction of the 24-hr clock eliminates the synchronizer at 1800. However, almost at once, activity, eating, and drinking are kept together by a second synchronizer, start of the light (by the electric clock at 0600). This may explain the ability of the rat to survive after destruction of the 24-hr clock.

Animals↗