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R Klinke

Publications and source records attributed to R Klinke.

At least 19 recordsLinked to original sources

Postnatal cortical development in congenital auditory deprivation.

The study investigates early postnatal development of local field potentials (LFPs) in the primary auditory cortex of hearing and congenitally deaf cats. In hearing cats, LFPs elicited by electrical intracochlear stimulation demonstrated developmental changes in mid-latency range, including reductions in peak and onset latencies of individual waves and a maturation of their shape and latencies during the first 2 months of life. In long latency range (>80 ms), the P(1)/N(1) response appeared after the fourth week of life and further increased in amplitude and decreased in latency, reaching mature shapes between the fourth and sixth months after birth (p.n.). Cortical activated areas became increasingly smaller during the first 3 months of life, reaching mature values at the fourth month p.n. The layer-specific pattern of synaptic activity matured 4 months p.n. In congenitally deaf cats, the developmental pattern was different. The lowest cortical LFP thresholds were significantly smaller than in hearing controls, demonstrating a "hypersensitivity" to sensory inputs. The development of N(b) waves was delayed and altered and the long latency responses became smaller than in controls at the second and third months. The activated areas remained smaller than in controls until the third month, then they increased rapidly and exceeded the activated areas of age-matched controls. From the fourth month on, the activated areas decreased again and smaller synaptic currents were found in deaf cats than in controls. The presented data demonstrate that functional development of the auditory cortex critically depends on auditory experience.

Acoustic Stimulation↗

Absence of cross-modal reorganization in the primary auditory cortex of congenitally deaf cats.

To investigate possible cross-modal reorganization of the primary auditory cortex (field A1) in congenitally deaf cats, after years of auditory deprivation, multiunit activity and local field potentials were recorded in lightly anesthetized animals and compared with responses obtained in hearing cats. Local field potentials were also used for current source-density analyses. For visual stimulation, phase-reversal gratings of three to five different spatial frequencies and three to five different orientations were presented at the point of central vision. Peripheral visual field was tested using hand-held stimuli (light bar-shaped stimulus of different orientations, moved in different directions and flashed) typically used for neurophysiological characterization of visual fields. From 200 multiunit recordings, no response to visual stimuli could be found in A1 of any of the investigated animals. Using the current source-density analysis of local field potentials, no local generators of field potentials could be found within A1, despite of the presence of small local field potentials. No multiunit responses to somatosensory stimulation (whiskers, face, pinna, head, neck, all paws, back, tail) could be obtained. In conclusion, there were no indications for a cross-modal reorganization (visual, somatosensory) of area A1 in congenitally deaf cats.

Animals↗

Hearing after congenital deafness: central auditory plasticity and sensory deprivation.

The congenitally deaf cat suffers from a degeneration of the inner ear. The organ of Corti bears no hair cells, yet the auditory afferents are preserved. Since these animals have no auditory experience, they were used as a model for congenital deafness. Kittens were equipped with a cochlear implant at different ages and electro-stimulated over a period of 2.0-5.5 months using a monopolar single-channel compressed analogue stimulation strategy (VIENNA-type signal processor). Following a period of auditory experience, we investigated cortical field potentials in response to electrical biphasic pulses applied by means of the cochlear implant. In comparison to naive unstimulated deaf cats and normal hearing cats, the chronically stimulated animals showed larger cortical regions producing middle-latency responses at or above 300 microV amplitude at the contralateral as well as the ipsilateral auditory cortex. The cortex ipsilateral to the chronically stimulated ear did not show any signs of reduced responsiveness when stimulating the 'untrained' ear through a second cochlear implant inserted in the final experiment. With comparable duration of auditory training, the activated cortical area was substantially smaller if implantation had been performed at an older age of 5-6 months. The data emphasize that young sensory systems in cats have a higher capacity for plasticity than older ones and that there is a sensitive period for the cat's auditory system.

Age Factors↗

Biophysical characterization of a soluble CD40 ligand (CD154) coiled-coil trimer: evidence of a reversible acid-denatured molten globule.

The CD40 ligand molecule is unique, consisting of a receptor-binding domain anchored by an isoleucine zipper moiety. Exact determination of the multimeric state and its tendency to form molten globules has not been elucidated. Corroborating evidence of a trimerized molecule in aqueous solution was obtained from size-exclusion chromatography, laser light scattering, and analytical ultracentrifugation. A reversible acid-denatured molten globule state was observed from circular dichroism and fluorescence spectroscopy data. The molten globule state was characterized by a loss of tertiary structure with associated retention of secondary structure near pH 3. Once returned to pH 7, the acid-denatured state refolded over the course of 7 days resulting in approximately 90% recovery of the native structure. The molten globule state was characterized by a broadening of structural features in the second-derivative spectra of Fourier transform infrared spectroscopy. A component band at 1650 cm(-1) was shown to be alpha-helix and originate from amide carbonyl vibrations of the isoleucine zipper. Differential scanning calorimetry measurements characterized the pH-sensitive molten globule state at pH 3.3 as one lacking a well-defined unfolding transition with an accompanying baseline shift at 58 degrees C (a consequence of increased heat capacity). The tendency to form molten globules during acid denaturation stress permits an opportunity to study the process of partial protein unfolding with implications concerning stability. Although reversible molten globules can be formed, it is important to recognize the unusual nature since the molten globule state is formed exclusively within the beta-sheet receptor-binding region.

Animals↗

Congenital auditory deprivation reduces synaptic activity within the auditory cortex in a layer-specific manner.

The present study investigates the functional deficits of naive auditory cortices in adult congenitally deaf cats. For this purpose, their auditory system was stimulated electrically using cochlear implants. Synaptic currents in cortical layers were revealed using current source density analyses. They were compared with synaptic currents found in electrically stimulated hearing cats. The naive auditory cortex showed significant deficits in synaptic activity in infragranular cortical layers. Furthermore, there was also a deficit of synaptic activities at longer latencies (>30 ms). The 'cortical column' was not activated in the well-defined sequence found in normal hearing cats. These results demonstrate functional deficits as a consequence of congenital auditory deprivation. Similar deficits are likely in congenitally deaf children.

Animals↗

[Determinants of the effectiveness of electric stimulation of the auditory nerve with cochlear implants: II. Configuration of the stimulating electrodes].

The influence of the electrode configuration on the efficacy of the electrical stimulation were investigated in the auditory nerve. For this purpose normal hearing cats were deafened by intrascalar application of neomycin and stimulated by a NUCLEUS-22 implant. Stimulation was performed in monopolar, bipolar and tripolar mode with all electrodes of the implant. The smallest thresholds were determined for monopolar stimulation, followed by tripolar and bipolar stimulation. The most focused stimulation was achieved by tripolar stimulation, followed by bipolar and monopolar stimulation. Tripolar stimulation is therefore suitable for beam forming in electrical stimulation of the auditory nerve.

Animals↗

[Determinants of the effectiveness of cochlear implants: I. Placement in the cochlea].

The present paper introduces a registration method for the electrical field produced by a cochlear implant in the scala tympani in vitro. It was possible to determine the electrical potentials with high spatial resolution using an additional recording electrode. The results indicate that the efficacy of the stimulation by the more apical electrodes is higher than by the more basal ones. The reason is the decreasing cross-sectional area of the scala tympani, which represents a decreasing shunt between the stimulation electrodes.

Animals↗

Recruitment of the auditory cortex in congenitally deaf cats by long-term cochlear electrostimulation.

In congenitally deaf cats, the central auditory system is deprived of acoustic input because of degeneration of the organ of Corti before the onset of hearing. Primary auditory afferents survive and can be stimulated electrically. By means of an intracochlear implant and an accompanying sound processor, congenitally deaf kittens were exposed to sounds and conditioned to respond to tones. After months of exposure to meaningful stimuli, the cortical activity in chronically implanted cats produced field potentials of higher amplitudes, expanded in area, developed long latency responses indicative of intracortical information processing, and showed more synaptic efficacy than in naïve, unstimulated deaf cats. The activity established by auditory experience resembles activity in hearing animals.

Acoustic Stimulation↗

Monitoring of anaesthesia in neurophysiological experiments.

Cortical activity can be substantially changed by the type of anaesthetic used, and by its dose level. For easy monitoring of depth of anaesthesia we describe the changes in electroencephalogram and electrocardiogram accompanying changes in depth of anaesthesia in the cat. Anaesthesia was induced by the volatile anaesthetic isoflurane. The high-frequency components (around 30 Hz) in the electroencephalogram disappear in deep anaesthesia. The electrocardiogram also shows substantial changes in contamination due to muscle fasciculations with anaesthesia level. Fasciculations appear as noise in the electrocardiogram. The amplitude of the electrical muscle activity contaminating the ECG can be easily used for the maintainance of a constant level of anaesthesia during a neurophysiological experiment.

Anesthesia↗

Incorporation of an isoleucine zipper motif enhances the biological activity of soluble CD40L (CD154).

Recent progress in the understanding of immune function indicates that the interaction of CD40L with its receptor, CD40, plays a pivotal role in both humoral immunity and cell-mediated defense against pathogens. Functional studies of this interaction on both dendritic cells and malignant cells have demonstrated that CD40L also plays an important role in immune surveillance and anti-tumor immunity. CD40L exists in nature predominantly as a membrane-anchored molecule. To develop CD40L as a potential therapeutic, it is important to optimize soluble forms of this molecule that could be used in a clinical setting. Several reports have shown that soluble forms of CD40L, like CD40 antibodies, are biologically active. In the present report we demonstrate that the incorporation of an isoleucine zipper trimerization motif significantly enhances the biological activity of soluble CD40L.

Animals↗

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↗

A stimulator with wireless power and signal transmission for implantation in animal experiments and other applications.

Functional electrical stimulation in alert experimental animals is often hampered, as free mobility of the animals is limited by the leads. On the other hand, portable devices carrying the power supply are not always accepted by the animal. For wireless transmission of power and stimulus code a system designed for implantation is described below. The power for the implant is supplied inductively by a rotating magnetic field. The stimulation signal is radio-transmitted by FM in the 140 MHz range and processed by the implant. Finally, a current source is driven for electrical stimulation. The present design is intended to be used for electrical cochlear stimulation. However, the circuit can also be used for other types of functional electrical stimulation.

Animals↗

A model for prelingual deafness, the congenitally deaf white cat--population statistics and degenerative changes.

Cochlear implantation in congenitally deaf children leads to electrical stimulation of an entirely naive central auditory system. In this case, processes of central auditory maturation are induced by the electric stimuli. For the study of these processes the deaf white cat (DWC) appears to be an appropriate model. However, a knowledge of the basic data of these animals is necessary before such a model may be used. This paper presents these data and is one of a series of publications concerning congenital deafness in children and cochlear implantation. In our strain 72% of the animals are totally deaf as judged by the absence of any brain stem evoked potentials at click intensities up to 120 dB SPL peak equivalent. Primarily, there is a degeneration of the entire organ of Corti during the first postnatal weeks. An absence of acoustically evoked brain stem responses in the early postnatal weeks shows that DWCs probably never have any hearing experience. Months after the degeneration of the organ of Corti, the spiral ganglion starts to degenerate from the midportion of the cochlea. However, even in adult cats (2 years), a sufficient number of functionally intact auditory afferents remain, which are suitable for electrical cochlear stimulation.

Acoustic Stimulation↗

Hair cell loss and regeneration after severe acoustic overstimulation in the adult pigeon.

The extent of hair cell regeneration following acoustic overstimulation severe enough to destroy tall hair cells, was determined in adult pigeons. BrdU (5-bromo-2'-deoxyuridine) was used as a proliferation marker. Recovery of hearing thresholds in each individual animal was measured over a period of up to 16 weeks after trauma. In ears with loss of both short and tall hair cells, little or no functional recovery occurred. In ears with less damage, where significant functional recovery did occur, there were always a few rows of surviving hair cells left at the neural edge of the basilar papilla. In the region of hair cell loss, numerous BrdU labeled cells were found. However, only a small minority of these cells were regenerated hair cells, the majority being monolayer cells. Irrespective of the extent of the region of hair cell loss, regenerated hair cells were observed predominantly in a narrow strip at the transition from the abneural area of total hair cell loss and the neural area of hair cell survival. With increasing damage this strip moved progressively towards the neural edge of the papilla. No regeneration of hair cells was observed in the abneural region of total hair cell loss, even up to 16 weeks after trauma. The results indicate that there is a gradient in the destructive effect of loud sound across the width of the basilar papilla, from most detrimental at the abneural edge to least detrimental at the neural edge. Both tall and short hair cells can regenerate after sound trauma. Whether they do regenerate or not depends on the degree of damage to the area of the papilla where they normally reside. Regeneration of new hair cells occurs only in a narrow longitudinal band, which moves from abneural into the neural direction with increasing damage. In the area neural to this band, hair cells survive the overstimulation. In the area abneural to this band, sound damage is so severe, that no regeneration of hair cells occurs. As a consequence morphological and functional deficits persist.

Acoustic Stimulation↗

Spatial resolution of cochlear implants: the electrical field and excitation of auditory afferents.

This paper investigates the spatial resolution of electrical intracochlear stimulation in order to enable further refinement of cochlear implants. For this purpose electrical potential distributions around a conventional human intracochlear electrode (NUCLEUS-22) were measured in a tank, in cat cadaver cochleae and in living cat cochleae. Potential gradients were calculated where of importance. The values were compared to spatial tuning curves from cat primary auditory afferents in electrical mono-, bi-, and various tripolar stimulation modes. Finally, a lumped element model was developed to elucidate the single fiber data. Tank potential measurements show the principal features of the different stimulation modes but are not sufficient to explain all the features of experimental data from single fibers. Intracochlear potential measurements indicate an increase in spatial resolution in an apical direction. The single fiber data also confirm that a tripolar stimulus configuration provides significantly better spatial resolution than any other stimulation mode presently in use.

Animals↗

A versatile system for the generation and the development of speech coding strategies in cochlear implants.

A signal processor-based (DSP32C) stimulation system for cochlear implants has been developed. This system allows up to 14 stimulation channels (place principle) and conserves the time structure of the acoustic signal (periodicity principle). Any combination of these features is possible; in compressed analogue mode, pulsatile mode, or combinations of both. The system also takes electrical crosstalk into account caused by the electrical properties of the inner ear and compensates for it. Lateral inhibition strategies improve spatial resolution. Finally, the system offers an interface for the generation of quick continuous interleaved sampling strategies (CIS); up to 11,000 pulses/s per channel.

Cochlear Implants↗

Expression of trimeric CD40 ligand in Pichia pastoris: use of a rapid method to detect high-level expressing transformants.

Pichia pastoris is a yeast capable of expressing large amounts of some proteins. When expression vectors are introduced into P. pastoris, individual transformants typically express widely varying amounts of protein. Because clones expressing the highest level of protein occur infrequently during the transformation process, finding them can be very labor-intensive. We developed an immunological based filter screening method that rapidly detects transformants secreting large amounts of a heterologous protein. We have applied this method to the expression of a soluble trimeric form of CD40L, a molecule that regulates B-cell responses. Using this method, we identified transformants with one to 13 copies of the CD40L expression cassette. Maximum expression was obtained with clones containing eight or more copies of the expression cassette, and a clone with eight copies was selected for further analysis. High cell density fermentation of this clone using a mixed glycerol:methanol feed yielded 255 mg CD40L per liter of supernatant.

Amino Acid Sequence↗

Afferent projection patterns in the auditory brainstem in normal and congenitally deaf white cats.

Cochlear implantation in congenitally deaf children is developing to a successful medical tool. Little is known, however, on morphology and pathophysiology of the central auditory system in these auditory deprived children. One form of congenital hearing loss, that seen in the deaf white cat, was investigated to see if there are differences in the afferent pathways from the cochlear nuclei to the inferior colliculus. The retrogradely transported fluorescent tracer diamidino yellow (DY) was injected into different parts of the central nucleus of the inferior colliculus (ICC) of normal cats and deaf white cats. It was found that the main afferent projection patterns in deaf white cats were unchanged in spite of congenital auditory deprivation; minor differences were seen.

Amidines↗