Stapedius muscle fibre characterization in the noise exposed and auditory deprived rat.

In skeletal muscle, interventions that unload the muscle cause slow-to-fast myosin heavy chain (MHC) conversions, whereas fast-to-slow conversions are seen when the muscles are engaged in resistance training and endurance exercise. The stapedius muscle (SM) is reported to prevent cochlear damage by noise. This theory may be supported by showing comparable changes of muscle fibre composition when ears are exposed to longstanding noise (SM training). Comparable changes after sound deprivation (SM unloading) would suggest that the SM needs a certain degree of daily activity evoked by environmental sound to sustain its normal composition. We investigated the difference in myosin composition of SM fibres from rats exposed to noise, from auditory deprived rats and from rats exposed to low level ambient noise (control group). Consecutive complete SM cross-sections were processed by enzymehistochemistry to determine acid/alkali lability of myofibrillar adenosine triphosphatase (mATPase) and by immunohistochemistry using MHC antibodies. Fibres were assigned to mATPase type I, IIA, IIX or 'Miscellaneous' categories. Per mATPase category, the fibres were attributed to groups with specific MHC isoform compositions. Auditory deprivation lasting nine weeks was accomplished by closure of the external meatus at the age of three weeks. A slow-to-fast shift was seen in these rats when compared to the control group. The noise exposed group was exposed to 65-90dB sound pressure level during a period lasting nine weeks from the age of three weeks onwards. A shift from an overwhelming presence of type mATPase IIX, as seen in the control group, to type mATPase IIA occurred in the noise exposed group. Also, more MHC IIA/IIX hybrid fibres were found in the mATPase IIX category. An adaptive response to the acoustic environment in the characteristics of the fibres of the SM, comparable to the response in skeletal muscles on unloading and training activity, can be ascertained. This supports the theory that the SM plays an active role in modulating external acoustic energy on entry to the cochlea. Our results are also in favour of another postulated function of the SM, the unmasking of high-frequency signals in low-frequency background noise.

Stapedius muscle fiber characterization during postnatal development in the rat.

The stapedius muscle (SM) is reported to prevent cochlear damage by noise. Functional demands are then the ability of fast contraction with long endurance. At the end of the third postnatal week, the middle ear of the rat is completely pneumatized and according to electrophysiological data, the auditory function starts to match the adult. We investigated the developmental changes in myosin composition of SM fibres using consecutive complete SM cross-sections (taken from rats on post natal day (PND) 7, 14, 16, 21, 28, 42 and 84) which were processed by enzymehistochemistry to determine acid/alkali lability of myofibrillar adenosine triphosphatase (mATPase) and by immunohistochemistry using myosin heavy chain (MHC) antibodies (mAb). Fibres were assigned to mATPase type I, IIA, IIB, IIX or 'Miscellaneous' categories. Per mATPase category, the fibres were attributed to groups with specific MHC isoform compositions. Neonatal MHC expression could not be documented with the mAb used. However, embryonal (Emb) MHC was expressed at PND 7, very little at PND 14; at later PND fibres did not show Emb MHC. In general, the mATPase-based classification did not show large alterations after PND 21. Expression of MHC IIB, which was present in almost 50% of the fibres at PND 7 and 14, diminished to 3% at PND 84. A decrease in number of fibres expressing more than one MHC isoform was found. These results show that the SM is a precociously developing muscle compared to limb muscles and even to the diaphragm. Moreover, it is shown that the expression of the adult MHC isoform phenotype coincides with the onset of auditory function in the third postnatal week.

Effects of isoproterenol on the orbicularis oculi and stapedius muscles.

The effect of the beta-adrenoceptor agonist, isoproterenol, on the orbicularis oculi and stapedius muscles was studied. Rats were injected with isoproterenol (5 mg/kg) every day. The animals were sacrificed after 1 and 2 weeks, and the orbicularis oculi muscle and stapedius muscle were removed. Muscle fibers were separated into single fibers, and cytochrome oxidase enzyme staining was applied with 1% MnCl2, DAB + H2O2. The density of cytochrome oxidase enzyme reaction products and the diameter of individual muscle fibers were analyzed by computer-assisted measurement. The administration of isoproterenol increased the activity of cytochrome oxidase as well as the diameter of muscle fibers in the orbicularis oculi muscle and stapedius muscle. It was suggested that these muscle fibers exhibited increases in functional activity after the administration. The results showed that the diameter and cytochrome oxidase enzyme activity of the orbicularis oculi and stapedius muscles increased. We conclude that isoproterenol induces hypertrophy of the orbicularis oculi and the stapedius muscle fibers, and promotes cytochrome oxidase enzyme activity.

Contraction properties and functional morphology of the avian stapedius muscle.

The influence of the stapedius muscle contraction on middle ear volume and acoustic impedance was investigated in the chicken, Gallus gallus. The time course of twitch responses to electrical stimulation (measured as volume and impedance changes) was found to be largely independent of the stimulus voltage, having a contraction time of 22 ms and a half-relaxation time of 22 ms. The stapedius muscle was therefore characterized as a fast twitch muscle. Slow contraction properties were also revealed: A summation of responses to repetitive stimulation beginning at 2.5 Hz and a slow decline to baseline were seen in volume and impedance change recordings. The morphological characteristics were consonant with that of a homogeneously fast muscle: Only fibres with high ATPase activity were identified and no fibres with "en grappe" or multiple innervation were observed. The slow characteristics were suggested to be due to visco-elastic elements in the middle ear. The chicken stapedius muscle is suggested to be analogous to both the stapedius and the tensor tympani of mammals.