R Pereira1,2, M E Oliveira3,4, R Santos3,4,5, E Oliveira6,3, T Barbosa7, T Santos8, P Gonçalves8, L Ferraz9, S Pinto10, A Barros10,11, J Oliveira6,3,4, M Sousa6,3. 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. ruterpereira@gmail.com. 2. Multidisciplinary Unit for Biomedical Research (UMIB), ICBAS-UP, Porto, Portugal. ruterpereira@gmail.com. 3. Multidisciplinary Unit for Biomedical Research (UMIB), ICBAS-UP, Porto, Portugal. 4. 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. 5. 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. 6. 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. 7. Department of Pediatrics, Maternal Child Centre of the North (CMIN), University Hospital Centre of Porto (CHUP), Largo da Maternidade, 4050-371, Porto, Portugal. 8. 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. 9. Department of Urology, Hospital Centre of Vila Nova de Gaia/Espinho, Unit 1, Rua Conceição Fernandes 1079, 4434-502, Vila Nova de Gaia, Portugal. 10. Centre for Reproductive Genetics Prof. Alberto Barros (CGR), Av. do Bessa, 240, 1° Dto. Frente, 4100-012, Porto, Portugal. 11. Department of Genetics, Faculty of Medicine, University of Porto (FMUP), Alameda Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.
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.
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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