The Drosophila Homologue of Tubulin-Specific Chaperone E–Like Protein Is Required for Synchronous Sperm Individualization and Normal Male Fertility

Abstract

The tubulin-specific chaperone E–like protein (TBCEL or E-like) of vertebrates shows sequence homology to TBCE, a component of the multimolecular complex required for tubulin heterodimer formation in all eukaryotic cells. TBCEL apparently serves more specific functions, as it is found only in animals. At the cellular level, TBCEL plays a role as a regulator of tubulin stability. It is strongly expressed in human testes, but its systemic function is not known. The gene CG12214 codes for the Drosophila homologue of the vertebrate TBCEL protein. Here we show that disruption of the Drosophila Tbcel gene causes defects in spermatid individualixation, which leads to dispersed migration of F-actin–rich investment cones. Mutations affecting the Tbcel gene cause strong reduction in male, but not female, fertility. However, mature sperm function apparently is not impaired. We generated polyclonal antisera against TBCEL to study its localization and distribution in Drosophila tissues. Immunostainings of wild-type and null mutant testes demonstrated that TBCEL is localized in testes, presumably associated with axoneme bundles prior to spermatid individualization. Molecular analysis of the transposon insertion site in the mutant mulet (mlt), for which male sterility and sperm individualization defects have previously been described, demonstrates that the mlt P-element insertion resides in the Tbcel gene. Our results show that loss of TBCEL in Drosophila is compatible with viability and normal female fertility but causes reduced male fertility. We conclude that Drosophila TBCEL is strongly expressed in testes and plays an important role in sperm individualization during spermatogenesis. The high level of Tbcel mRNA in human testes suggests a general role of TBCEL in animal spermatogenesis. However, Western blots and courtship analysis suggest that TBCEL may have additional functions in the nervous system of Drosophila that could contribute to the observed reduced male fertility. These functions now have to be investigated.

Keywords::

• antiserum
• axonemes
• courtship
• E-like, male sterility
• TBCE-like
• TBCEL
• testes

ACKNOWLEDGMENTS

We would like to thank Prof. Georg Krohne (Electron Microscopy Unit, Biocenter, University of Würzburg) for generating the anti-TBCEL antisera. We are grateful to the Bloomington, Exelixis, and Kyoto stock centers for providing fly stocks and other resources, and to Gertrud Gramlich and Kathrin Fink for technical help.

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

This work was supported by grants from the Deutsche Forschungsgemeinschaft to EB (SFB554, SFB581, GRK1156).