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Transition zone assembly and its contribution to axoneme formation in Drosophila male germ cells

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ABSTRACT

Ciliary transition zone (TZ) assembly is complex and incompletely understood. Vieillard et al. show that Drosophila Cby and Dila cooperate to assemble the TZ and membrane cap, which, together with microtubule remodeling by kinesin-13, is required for axoneme formation in male germ cells.

No MeSH data available.


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Cby and Dila cooperate to build the TZ and ciliary cap in male germ cells. (A) Whole-mount Drosophila testis expressing the tagged mitochondria protein DonJuan::GFP observed by bright-field and confocal microscopy. The cysts fail to elongate in dila81; cby1 mutants. (B–D) Confocal imaging of centrioles/BB of squashed testes. (B) Detection of Mks1::GFP or B9d1 (green) and Asterless or γ-tubulin (red), respectively. Mks1 and B9d1 are absent from the tip of the centrioles in spermatocytes and from the tip of BB in spermatids of dila81; cby1 mutants. (C) Cep290::GFP (green) and Asterless (red) staining. Cep290 localization at the tip of the centrioles in spermatocytes and at the tip of BB in spermatids is almost completely lost in dila81; cby1 mutants. (D) No significant differences in Unc::mkate2 localization is observed at early spermatocyte stages. Unc::mKate2 localization is expanded in dila81; cby1 mutants from midspermatocyte stage and in spermatids (arrows). Bars: (A) 50 µm; (B) 5 µm; (C and D) 2 µm.
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fig6: Cby and Dila cooperate to build the TZ and ciliary cap in male germ cells. (A) Whole-mount Drosophila testis expressing the tagged mitochondria protein DonJuan::GFP observed by bright-field and confocal microscopy. The cysts fail to elongate in dila81; cby1 mutants. (B–D) Confocal imaging of centrioles/BB of squashed testes. (B) Detection of Mks1::GFP or B9d1 (green) and Asterless or γ-tubulin (red), respectively. Mks1 and B9d1 are absent from the tip of the centrioles in spermatocytes and from the tip of BB in spermatids of dila81; cby1 mutants. (C) Cep290::GFP (green) and Asterless (red) staining. Cep290 localization at the tip of the centrioles in spermatocytes and at the tip of BB in spermatids is almost completely lost in dila81; cby1 mutants. (D) No significant differences in Unc::mkate2 localization is observed at early spermatocyte stages. Unc::mKate2 localization is expanded in dila81; cby1 mutants from midspermatocyte stage and in spermatids (arrows). Bars: (A) 50 µm; (B) 5 µm; (C and D) 2 µm.

Mentions: In double dila81; cby1 mutant flies, in addition to strong sensory defects, males are completely sterile. No mature sperm is produced and examination of testes shows that sperm cysts failed to elongate whereas the overall size of the testes is not reduced (Fig. 6 A). EM examination of testes indicates a severe disorganization of sperm cysts (Fig. S3 A). Most axonemes were absent or completely damaged. Because cysts were severely disorganized, we could only count the number of remaining axonemes relative to the number of major mitochondria derivatives. Out of 442 mitochondria, only 5 intact axonemes (1.2%) and 28 recognizable but destroyed axonemes (6.3%; Fig. S3 A) could be identified. These observations indicate a strong genetic interaction between cby and dila in flagella formation, as we observed only a small fraction (14%) of defective flagellar axonemes in cby1 mutants (Enjolras et al., 2012) and no axonemal defects were described in dila81 mutants (Ma and Jarman, 2011).


Transition zone assembly and its contribution to axoneme formation in Drosophila male germ cells
Cby and Dila cooperate to build the TZ and ciliary cap in male germ cells. (A) Whole-mount Drosophila testis expressing the tagged mitochondria protein DonJuan::GFP observed by bright-field and confocal microscopy. The cysts fail to elongate in dila81; cby1 mutants. (B–D) Confocal imaging of centrioles/BB of squashed testes. (B) Detection of Mks1::GFP or B9d1 (green) and Asterless or γ-tubulin (red), respectively. Mks1 and B9d1 are absent from the tip of the centrioles in spermatocytes and from the tip of BB in spermatids of dila81; cby1 mutants. (C) Cep290::GFP (green) and Asterless (red) staining. Cep290 localization at the tip of the centrioles in spermatocytes and at the tip of BB in spermatids is almost completely lost in dila81; cby1 mutants. (D) No significant differences in Unc::mkate2 localization is observed at early spermatocyte stages. Unc::mKate2 localization is expanded in dila81; cby1 mutants from midspermatocyte stage and in spermatids (arrows). Bars: (A) 50 µm; (B) 5 µm; (C and D) 2 µm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5037411&req=5

fig6: Cby and Dila cooperate to build the TZ and ciliary cap in male germ cells. (A) Whole-mount Drosophila testis expressing the tagged mitochondria protein DonJuan::GFP observed by bright-field and confocal microscopy. The cysts fail to elongate in dila81; cby1 mutants. (B–D) Confocal imaging of centrioles/BB of squashed testes. (B) Detection of Mks1::GFP or B9d1 (green) and Asterless or γ-tubulin (red), respectively. Mks1 and B9d1 are absent from the tip of the centrioles in spermatocytes and from the tip of BB in spermatids of dila81; cby1 mutants. (C) Cep290::GFP (green) and Asterless (red) staining. Cep290 localization at the tip of the centrioles in spermatocytes and at the tip of BB in spermatids is almost completely lost in dila81; cby1 mutants. (D) No significant differences in Unc::mkate2 localization is observed at early spermatocyte stages. Unc::mKate2 localization is expanded in dila81; cby1 mutants from midspermatocyte stage and in spermatids (arrows). Bars: (A) 50 µm; (B) 5 µm; (C and D) 2 µm.
Mentions: In double dila81; cby1 mutant flies, in addition to strong sensory defects, males are completely sterile. No mature sperm is produced and examination of testes shows that sperm cysts failed to elongate whereas the overall size of the testes is not reduced (Fig. 6 A). EM examination of testes indicates a severe disorganization of sperm cysts (Fig. S3 A). Most axonemes were absent or completely damaged. Because cysts were severely disorganized, we could only count the number of remaining axonemes relative to the number of major mitochondria derivatives. Out of 442 mitochondria, only 5 intact axonemes (1.2%) and 28 recognizable but destroyed axonemes (6.3%; Fig. S3 A) could be identified. These observations indicate a strong genetic interaction between cby and dila in flagella formation, as we observed only a small fraction (14%) of defective flagellar axonemes in cby1 mutants (Enjolras et al., 2012) and no axonemal defects were described in dila81 mutants (Ma and Jarman, 2011).

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Ciliary transition zone (TZ) assembly is complex and incompletely understood. Vieillard et al. show that Drosophila Cby and Dila cooperate to assemble the TZ and membrane cap, which, together with microtubule remodeling by kinesin-13, is required for axoneme formation in male germ cells.

No MeSH data available.


Related in: MedlinePlus