Zusammenfassung
Diese Arbeit vergleicht die embryonale Entwicklung von Adenylatzyklase und die Markierung von Phospholipiden im Innenohr in vivo und in vitro. Die Innenohranlage von CBA-Mäusen wurde am 16. Tag der Entwicklung explantiert und 3–12 Tage in vitro kultiviert. Die Gestationszeit in vivo beträgt 21 Tage.
Während der ersten Woche wächst das Innenohr in vitro nur wenig, gemessen am Proteingehalt. Während der gleichen Zeit erhöht sich der Proteingehalt des Labyrinthes in vivo etwa dreifach.
Die Aktivität der Adenylatzyklase ist vergleichbar in vivo und in vitro bis zum 19. Tag der Entwicklung. Danach übersteigt die spezifische Aktivität des Enzymes in vitro die Aktivität in vivo um das drei- bis fünffache, was auf einen Mangel an Kontrollmechanismen schließen läßt.
Phospholipide werden mit 32P in vivo und in vitro qualitativ ähnlich markiert, obwohl auch hier quantitative Unterschiede bestehen.
Nach dieser Untersuchung scheint die Bedeutung der Organkultur für Entwicklungsstudien auf die Zeit vor der Geburt beschränkt zu sein.
Summary
Adenylate cyclase activity and phospholipid labeling were compared during embryonic development of the mouse inner ear in vivo and in vitro. Inner ears were explanted on the 16th gestational day and cultured in vitro for 3–12 days. The gestation time in vivo is 21 days.
During the 1st week in vitro there is very little growth of the inner ear with regard to total protein content. In contrast, the labyrinth increases its protein content threefold during the corresponding period of time in vivo.
The activity of adenylate cyclase develops parallel in vivo and in vitro until the 19th gestational day whereafter the specific activity of the enzyme in vitro surpasses that of the enzyme in vivo three- to fivefold suggesting a lack of control mechanisms in organ culture. Phospholipids are labeled by 32P in an essentially similar quantitative relationship in vivo and in vitro, while some quantitative differences exist.
According to the present study the usefulness of the organ culture for the investigation of inner ear development appears limited to a culture period corresponding to an age prior to birth.
This is a preview of subscription content, log in via an institution to check access.
References
Anniko M, Van De Water TR, Nordemar H (1978) Organ culture of the 16th gestation day mouse labyrinth. A model suggestion for pre- and post-partum development. Acta Otolaryngol (Stockh) 86: 52–55
Anniko M, Nordemar H, Van De Water TR (1979) Embryogenesis of the inner ear. I. Development and differentiation of the inner ear. I. Development and differentiation of the mammalian crista ampullaris in vivo and in vitro. Arch Otorhinolaryngol (NY) 224: 285–299
Anniko M (1980) Embryogenesis of the inner ear. III. Formation of the tectorial membrane in vivo and in vitro. Anat Embryol (Berl) 160: 301–313
Anniko M, Nordemar H (1980) Embryogenesis of the inner ear. IV. Post-natal maturation of the secretory epithelia of the inner ear in correlation with the elemental composition in the endolymphatic space. Arch Otorhinolaryngol (NY) 229: 281–288
Bagger-Sjöbäck D, Filipek CS, Schacht J (1980) Characteristics and drug responses of cochlear and vestibular adenylate cyclase. Arch Otorhinolaryngol (NY) 228: 217–222
Li CW, Van De Water TR, Ruben RJ (1976) In vitro study of fate mapping of the mouse otocyst. Trans Am Acad Ophthalmol Otolaryngol 82: 273–280
Lodhi S, Weiner ND, Mechigian I, Schacht J (1980) Ototoxicity of aminoglycosides correlated with their action on monomolecular films of polyphosphoinositides. Biochem Pharmacol 29: 597–601
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275
Schacht J (1978) Purification of polyphosphoinositides by chromatography on immobilized neomycin. J Lipid Res 19: 1063–1067
Schacht J (1979) Isolation of an aminoglycoside receptor from guinea pig inner ear tissues and kidney. Arch Otorhinolaryngol (NY) 224: 129–134
Steiner AL (1974) Assay of cyclic nucleotides by radioimmunoassay methods. In: Harman JG, O'Malley BW (eds) Methods in enzymology, vol 38. Academic Press, New York, pp 96–105
Stockhorst E, Schacht J (1977) Radioactive labeling of phospholipids and proteins by cochlear perfusion in the guinea pig and the effects of neomycin. Acta Otolaryngol (Stockh) 83: 401–409
Van De Water TR, Ruben RJ (1971) Organ culture of the mammalian inner ear. Acta Otolaryngol (Stockh) 71: 303–312
Van De Water TR, Ruben RJ (1973) Quantification of the in vitro development of the mouse embryo inner ear. Ann Otol Rhinol Laryngol [Suppl 3] 82: 19–21
Van De Water TR (1976) Effects of removal of the statoacoustic ganglian complex upon the growing otocyst. Ann Otol Rhinol Laryngol [Suppl 33] 85: 1–32
Author information
Authors and Affiliations
Additional information
Supported by grants from Karolinska Institutet, The Foundation Tysta Skolan, The Torsten and Ragnar Söderberg Foundation and The Swedish Medical Research Council (12X-720) (Matti Anniko and Hans Nordemar). Also supported by research grant NS-12881, Program Project Grant NS-05785, and a Senior International Fellowship (FO6-TW00290) of the Fogarty International Center, National Institute of Health (Jochen Schacht)
Rights and permissions
About this article
Cite this article
Anniko, M., Nordemar, H., Spångberg, M.L. et al. Biochemical studies on the embryonic development of the mammalian inner ear in organ culture. Arch Otorhinolaryngol 230, 237–243 (1981). https://doi.org/10.1007/BF00456324
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00456324
Schlüsselwörter
- Innenohr
- Embryonale Entwicklung
- Organkultur
- Vergleich in vivo/in vitro
- Biochemie
- Protein
- Adenylatzyklase
- Phospholipide