Do the tensor tympani and tensor veli palatini muscles of man form a functional unit? A histochemical investigation of their putative connections.

The discussion among anatomists and otolaryngologists about the muscles originating from the Eustachian tube and the connections between the tensor tympani and tensor veli palatini muscles started in the 1860s. From then on, a considerable number of contradictory hypotheses and data have been presented. However, before discussing whether or not these two muscles form a functional unit, interest should focus on the question of whether it is even possible. The cartilaginous portion of the Eustachian tube with all muscles originating from it, including the whole tensor tympani muscle, was dissected from five perfusion-fixed cadavers and removed in toto. Complete longitudinal serial sections of 10 microm were made in the axis of the tensor tympani muscle. Sections were alternatingly stained according to Cason's and Maskar's techniques. The macroscopic aspect (under the operating microscope) of a tendinous connection between the two muscles under consideration could be proven by the histochemical methods used in all cases. Based on our findings and the literature reviewed we are convinced that the tensor tympani and tensor veli palatini muscles of man constitute a functional unit. This represents an important step forward towards the understanding of the possible functions the tensor tympani muscle serve in man.

Autosomal-dominant, prelingual, nonprogressive sensorineural hearing loss: localization of the gene (DFNA8) to chromosome 11q by linkage in an Austrian family.

A four-generation family suffering from an autosomal-dominant, congenital, nonprogressive, nonsyndromic hearing loss was found in a rural region of Austria. The hearing loss was moderate to severe, a pure tone audiogram showing a U-shaped form with maximum loss at 2, 000 Hz. An initial genome search led to a lod score of 3.01 with markers on chromosome 15. This locus was registered as DFNA8 in the HUGO data base. Further sampling of the family, however, yielded data that reduced the maximal lod score with chromosome 15 markers to 1.81. The genome search was restarted using an ABITM genotyper, which eventually detected several positive two-point lod scores with markers from the long arm of chromosome 11. The highest value was 3. 6, which was seen with the marker D11S934. Haplotype analysis excluded the gene from the chromosomal region proximal from D11S898 and distal to D11S1309. These results place the gene in the region of the hearing loss gene DFNA12. Recent evidence suggests that the somewhat different phenotypes found in these two families are due to two different mutations in the human alpha-tectorine gene (Verhoeven et al., 1998).

Mutations in the human alpha-tectorin gene cause autosomal dominant non-syndromic hearing impairment.

The tectorial membrane is an extracellular matrix of the inner ear that contacts the stereocilia bundles of specialized sensory hair cells. Sound induces movement of these hair cells relative to the tectorial membrane, deflects the stereocilia, and leads to fluctuations in hair-cell membrane potential, transducing sound into electrical signals. Alpha-tectorin is one of the major non-collagenous components of the tectorial membrane. Recently, the gene encoding mouse alpha-tectorin (Tecta) was mapped to a region of mouse chromosome 9, which shows evolutionary conservation with human chromosome 11q (ref. 3), where linkage was found in two families, one Belgian (DFNA12; ref. 4) and the other, Austrian (DFNA8; unpublished data), with autosomal dominant non-syndromic hearing impairment. We determined the complete sequence and the intron-exon structure of the human TECTA gene. In both families, mutation analysis revealed missense mutations which replace conserved amino-acid residues within the zona pellucida domain of TECTA. These findings indicate that mutations in TECTA are responsible for hearing impairment in these families, and implicate a new type of protein in the pathogenesis of hearing impairment.

Emergence of myogenic and endothelial cell lineages in avian embryos.

The roles of cell cycles and of cell-cell interactions in the emergence of myogenic and endothelial cell lineages were studied in avian embryos using the quail-chicken marker system. Quail embryos were treated with drugs preventing either DNA replication or the movement of cells. Portions of drug-treated or untreated quail blastoderms were grafted into chicken wing buds. After an incubation for an additional 4 to 10 days, the embryos were analyzed for the presence of quail muscle or quail endothelial cells by the Feulgen reaction and by immunostaining. Both cell lineages differ in the time of their commitment as well as in the conditions necessary for their emergence. Muscle cells did not differentiate from unincubated blastoderms nor did they develop from drug-treated blastoderms. These results corroborate that the commitment of myogenic cells occurs during gastrulation and indicate that this commitment requires both DNA replication and cellular movements allowing cell-cell and/or cell-matrix interactions. Endothelial cells, on the contrary, developed both from drug-treated and from unincubated blastoderms, indicating that their commitment occurs before and independent of gastrulation and does not require DNA replication during gastrulation.

Differentiation of endothelial cells in avian embryos does not depend on gastrulation.

Unincubated quail eggs were treated with Cytochalasin B. By this means, gastrulation of the blastodiscs was inhibited. Fragments of these blastodiscs were grafted into wings buds of chick embryos, and the differentiation fate of graft-derived cells was studied. Results show that only endothelial cells differentiate from the grafts. They were even found outside the graft site in vessels made up of a chimeric endothelium. It can be concluded that determination, differentiation and migration of endothelial cells does not depend on gastrulation.