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Characterization of CCDC103 expression profiles: further insights in primary ciliary dyskinesia and in human reproduction

  • Reproductive Physiology and Disease
  • Published:
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Abstract

Propose

To study CCDC103 expression profiles and understand how pathogenic variants in CCDC103 affect its expression profile at mRNA and protein level.

Methods

To increase the knowledge about the CCDC103, we attempted genotype-phenotype correlations in two patients carrying novel homozygous (missense and frameshift) CCDC103 variants. Whole-exome sequencing, quantitative PCR, Western blot, electron microscopy, immunohistochemistry, immunocytochemistry, and immunogold labelling were performed to characterize CCDC103 expression profiles in reproductive and somatic cells.

Results

Our data demonstrate that pathogenic variants in CCDC103 gene negatively affect gene and protein expression in both patients who presented absence of DA on their axonemes. Further, we firstly report that CCDC103 is expressed at different levels in reproductive tissues and somatic cells and described that CCDC103 protein forms oligomers with tissue-specific sizes, which suggests that CCDC103 possibly undergoes post-translational modifications. Moreover, we reported that CCDC103 was restricted to the midpiece of sperm and is present at the cytoplasm of the other cells.

Conclusions

Overall, our data support the CCDC103 involvement in PCD and suggest that CCDC103 may have different assemblies and roles in cilia and sperm flagella biology that are still unexplored.

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Acknowledgments

The authors would like to thank Ana Rita Gonçalves, MSc, Center of Medical Genetics Dr. Jacinto Magalhães (CGMJM-CHUP) for Sanger sequencing samples processing; Ângela Alves, MSc, Institute of Biomedical Sciences Abel Salazar-University of Porto (ICBAS-UP) for technical electron microscopy assistance; Raquel Bernardino, PhD, Tânia Dias, Msc, and Ana Maria, MSc for Sertoli cell culture assistance (ICBAS-UP); Fátima Ferreirinha, MSc (ICBAS-UP) for confocal microscopy assistance; and Rui Fernandes, MSc, Institute of Health Research and Innovation (IBMC/i3S-UP) for immunogold labelling assistance.

Funding

UMIB (Pest-OE/SAU/UI0215/2014) is funded by the National Funds through FCT-Foundation for Science and Technology, and FCT Grant ref.: PD/BD/105767/2014 (R.P.).

Author information

Author notes

  1. J. Oliveira and M. Sousa contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cell Biology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal

    R. Pereira, E. Oliveira, J. Oliveira & M. Sousa

  2. Multidisciplinary Unit for Biomedical Research (UMIB), ICBAS-UP, Porto, Portugal

    R. Pereira, M. E. Oliveira, R. Santos, E. Oliveira, J. Oliveira & M. Sousa

  3. Molecular Genetics Unit, Center of Medical Genetics Dr. Jacinto Magalhães (CGMJM), University Hospital Centre of Porto (CHUP), Praça Pedro Nunes, 88, 4099-028, Porto, Portugal

    M. E. Oliveira, R. Santos & J. Oliveira

  4. UCIBIO/REQUIMTE, Department of Biological Sciences, Laboratory of Biochemistry, Faculty of Pharmacy, University of Porto (FFUP), Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal

    R. Santos

  5. Department of Pediatrics, Maternal Child Centre of the North (CMIN), University Hospital Centre of Porto (CHUP), Largo da Maternidade, 4050-371, Porto, Portugal

    T. Barbosa

  6. Department of Otorhinolaryngology, S. Sebastião Hospital, Hospital Centre of entre Douro e Vouga, Rua Dr. Cândido Pinho 5, 4520-211, Santa Maria da Feira, Portugal

    T. Santos & P. Gonçalves

  7. Department of Urology, Hospital Centre of Vila Nova de Gaia/Espinho, Unit 1, Rua Conceição Fernandes 1079, 4434-502, Vila Nova de Gaia, Portugal

    L. Ferraz

  8. Centre for Reproductive Genetics Prof. Alberto Barros (CGR), Av. do Bessa, 240, 1° Dto. Frente, 4100-012, Porto, Portugal

    S. Pinto & A. Barros

  9. Department of Genetics, Faculty of Medicine, University of Porto (FMUP), Alameda Prof. Hernâni Monteiro, 4200-319, Porto, Portugal

    A. Barros

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  1. R. Pereira
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  2. M. E. Oliveira
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  4. E. Oliveira
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  7. P. Gonçalves
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  8. L. Ferraz
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  10. A. Barros
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  11. J. Oliveira
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Corresponding author

Correspondence to R. Pereira.

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Supplemental Video 1

Immunocytochemical detection of CCDC103 from control sperm (SZ) obtained from a confocal Z-stack. CCDC103 is present, with a helix-like shape, at sperm midpiece (green). Sperm flagella stained with axoneme-specific acetylated α-tubulin (red) and nuclei stained with DAPI (blue). (AVI 23 kb)

Supplementary Video 2

(AVI 27 kb)

Supplemental Table S1.

List of primers used in this study. (DOCX 16 kb)

Supplemental File S1.

Immunogold staining (DOCX 1360 kb)

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Pereira, R., Oliveira, M.E., Santos, R. et al. Characterization of CCDC103 expression profiles: further insights in primary ciliary dyskinesia and in human reproduction. J Assist Reprod Genet 36, 1683–1700 (2019). https://doi.org/10.1007/s10815-019-01509-7

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