Effects of Tcte1 knockout on energy chain transportation and spermatogenesis: implications for male infertility.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:38650655", "kind": "science-record", "revision": 5, "content_hash": "sha256:cbef7497de884ba0121aeeef1b904f0d0d4ccdf1b14edd76e54bf90f091b6373", "created_at": "2026-11-01T19:20:37Z", "updated_at": "2026-11-01T22:54:11Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Effects of Tcte1 knockout on energy chain transportation and spermatogenesis: implications for male infertility.", "doi": "10.1093/hropen/hoae020", "pmid": "38650655", "authors": ["Marta Olszewska", "Agnieszka Malcher", "Tomasz Stokowy", "Nijole Pollock", "Andrea J Berman", "Sylwia Budkiewicz", "Marzena Kamieniczna", "Hanna Jackowiak", "Joanna Suszynska-Zajczyk", "Piotr Jedrzejczak", "Alexander N Yatsenko", "Maciej Kurpisz"], "journal": "Human reproduction open", "year": 2024, "abstract": "STUDY QUESTION: Is the Tcte1 mutation causative for male infertility?\nSUMMARY ANSWER: Our collected data underline the complex and devastating effect of the single-gene mutation on the testicular molecular network, leading to male reproductive failure.\nWHAT IS KNOWN ALREADY: Recent data have revealed mutations in genes related to axonemal dynein arms as causative for morphology and motility abnormalities in spermatozoa of infertile males, including dysplasia of fibrous sheath (DFS) and multiple morphological abnormalities in the sperm flagella (MMAF). The nexin-dynein regulatory complex (N-DRC) coordinates the dynein arm activity and is built from the DRC1-DRC7 proteins. DRC5 (TCTE1), one of the N-DRC elements, has already been reported as a candidate for abnormal sperm flagella beating; however, only in a restricted manner with no clear explanation of respective observations.\nSTUDY DESIGN SIZE DURATION: Using the CRISPR/Cas9 genome editing technique, a mouse Tcte1 gene knockout line was created on the basis of the C57Bl/6J strain. The mouse reproductive potential, semen characteristics, testicular gene expression levels, sperm ATP, and testis apoptosis level measurements were then assessed, followed by visualization of N-DRC proteins in sperm, and protein modeling in silico. Also, a pilot genomic sequencing study of samples from human infertile males (n = 248) was applied for screening of TCTE1 variants.\nPARTICIPANTS/MATERIALS SETTING METHODS: To check the reproductive potential of KO mice, adult animals were crossed for delivery of three litters per caged pair, but for no longer than for 6 months, in various combinations of zygosity. All experiments were performed for wild-type (WT, control group), heterozygous Tcte1+/- and homozygous Tcte1-/- male mice. Gross anatomy was performed on testis and epididymis samples, followed by semen analysis. Sequencing of RNA (RNAseq; Illumina) was done for mice testis tissues. STRING interactions were checked for protein-protein interactions, based on changed expression levels of corresponding genes identified in the mouse testis RNAseq experiments. Immunofluorescence in situ staining was performed to detect the N-DRC complex proteins: Tcte1 (Drc5), Drc7, Fbxl13 (Drc6), and Eps8l1 (Drc3) in mouse spermatozoa. To determine the amount of ATP in spermatozoa, the luminescence level was measured. In addition, immunofluorescence in situ staining was performed to check the level of apoptosis via caspase 3 visualization on mouse testis samples. DNA from whole blood samples of infertile males (n = 137 with non-obstructive azoospermia or cryptozoospermia, n = 111 samples with a spectrum of oligoasthenoteratozoospermia, including n = 47 with asthenozoospermia) was extracted to perform genomic sequencing (WGS, WES, or Sanger). Protein prediction modeling of human-identified variants and the exon 3 structure deleted in the mouse knockout was also performed.\nMAIN RESULTS AND THE ROLE OF CHANCE: No progeny at all was found for the homozygous males which were revealed to have oligoasthenoteratozoospermia, while heterozygous animals were fertile but manifested oligozoospermia, suggesting haploinsufficiency. RNA-sequencing of the testicular tissue showed the influence of Tcte1 mutations on the expression pattern of 21 genes responsible for mitochondrial ATP processing or linked with apoptosis or spermatogenesis. In Tcte1-/- males, the protein was revealed in only residual amounts in the sperm head nucleus and was not transported to the sperm flagella, as were other N-DRC components. Decreased ATP levels (2.4-fold lower) were found in the spermatozoa of homozygous mice, together with disturbed tail:midpiece ratios, leading to abnormal sperm tail beating. Casp3-positive signals (indicating apoptosis) were observed in spermatogonia only, at a similar level in all three mouse genotypes. Mutation screening of human infertile males revealed one novel and five ultra-rare heterogeneous variants (predicted as disease-causing) in 6.05% of the patients studied. Protein prediction modeling of identified variants revealed changes in the protein surface charge potential, leading to disruption in helix flexibility or its dynamics, thus suggesting disrupted interactions of TCTE1 with its binding partners located within the axoneme.\nLARGE SCALE DATA: All data generated or analyzed during this study are included in this published article and its supplementary information files. RNAseq data are available in the GEO database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number GSE207805. The results described in the publication are based on whole-genome or exome sequencing data which includes sensitive information in the form of patient-specific germline variants. Information regarding such variants must not be shared publicly following European Union legislation, therefore access to raw data that support the findings of this study are available from the corresponding author upon reasonable request.\nLIMITATIONS REASONS FOR CAUTION: In the study, the in vitro fertilization performance of sperm from homozygous male mice was not checked.\nWIDER IMPLICATIONS OF THE FINDINGS: This study contains novel and comprehensive data concerning the role of TCTE1 in male infertility. The TCTE1 gene is the next one that should be added to the 'male infertility list' because of its crucial role in spermatogenesis and proper sperm functioning.\nSTUDY FUNDING/COMPETING INTERESTS: This work was supported by National Science Centre in Poland, grants no.: 2015/17/B/NZ2/01157 and 2020/37/B/NZ5/00549 (to M.K.), 2017/26/D/NZ5/00789 (to A.M.), and HD096723, GM127569-03, NIH SAP #4100085736 PA DoH (to A.N.Y.). The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.", "claims": [{"id": "claim-method", "statement": "Statistical analyses included the following: normality test (D’Agostino and Pearson or Kolmogorov–Smirnoff), unpaired two-tailed t-test with Welch’s correction or Mann–Whitney test, one-way ANOVA test and Fisher’s exact, and two-tailed Pearson correlation.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38650655", "locator": "sections[19] \"Statistical analysis\" paragraph 2", "quote": "Statistical analyses included the following: normality test (D’Agostino and Pearson or Kolmogorov–Smirnoff), unpaired two-tailed t-test with Welch’s correction or Mann–Whitney test, one-way ANOVA test and Fisher’s exact, and two-tailed Pearson correlation."}], "state": "complete"}, {"id": "claim-data-access", "statement": "All data generated or analyzed during this study are included in this published article and its supplemental information files. RNAseq data are available in GEO database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number: GSE207805. The results described in the publication are based on whole-genome or exome sequencing data which includes sensitive information in the form of patient-specific germline variants. Information regarding such variants must not be shared publicly following the European Union legislation described in the following document: https://publications.jrc.ec.europa.eu/repository/bitstream/JRC113479/policy_report_-_review_of_eu_national_legislation_on_genomics_-_with_identifiers_1.pdf, therefore the access to raw data that support findings of this study are available from the corresponding author upon reasonable request.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38650655", "locator": "sections[40] \"Data availability\" paragraph 2", "quote": "All data generated or analyzed during this study are included in this published article and its supplemental information files. RNAseq data are available in GEO database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number: GSE207805. The results described in the publication are based on whole-genome or exome sequencing data which includes sensitive information in the form of patient-specific germline variants. Information regarding such variants must not be shared publicly following the European Union legislation described in the following document: https://publications.jrc.ec.europa.eu/repository/bitstream/JRC113479/policy_report_-_review_of_eu_national_legislation_on_genomics_-_with_identifiers_1.pdf, therefore the access to raw data that support findings of this study are available from the corresponding author upon reasonable request."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:38650655"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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RNAseq data are available in GEO database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number: GSE207805. The results described in the publication are based on whole-genome or exome sequencing data which includes sensitive information in the form of patient-specific germline variants. Information regarding such variants must not be shared publicly following the European Union legislation described in the following document: https://publications.jrc.ec.europa.eu/repository/bitstream/JRC113479/policy_report_-_review_of_eu_national_legislation_on_genomics_-_with_identifiers_1.pdf, therefore the access to raw data that support findings of this study are available from the corresponding author upon reasonable request.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "All data generated or analyzed during this study are included in this published article and its supplemental information files. RNAseq data are available in GEO database (https://www.ncbi.nlm.nih.gov/geo/) under the accession number: GSE207805. The results described in the publication are based on whole-genome or exome sequencing data which includes sensitive information in the form of patient-specific germline variants. Information regarding such variants must not be shared publicly following the European Union legislation described in the following document: https://publications.jrc.ec.europa.eu/repository/bitstream/JRC113479/policy_report_-_review_of_eu_national_legislation_on_genomics_-_with_identifiers_1.pdf, therefore the access to raw data that support findings of this study are available from the corresponding author upon reasonable request.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM6319604", "GSM6319605", "GSM6319606", "GSM6319607", "GSM6319608", "GSM6319609", "GSM6319610", "GSM6319611", "GSM6319612", "GSM6319613", "GSM6319614"], "sample_count": 11, "group_sizes": null, "group_sizes_basis": null, "platform": "24247", "taxon": "Mus musculus", "title": "Tcte1 knockout influence on energy chain transportation, apoptosis and spermatogenesis – implications for male infertility", "resolution_checked_at": "2026-11-01T20:16:08Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:GSE207805 not downloaded", "asset:data:geo:GSE207805 licence not read"]}, "analysis": {"state": "complete", "methods": [{"id": "method-1", "family": "group_comparison", "specific": "welch_t_test", "label": "Group comparison", "dsl_operations": ["welch_t"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 1.0, "band": "verbatim", "quote": "Statistical analyses included the following: normality test (D’Agostino and Pearson or Kolmogorov–Smirnoff), unpaired two-tailed t-test with Welch’s correction or Mann–Whitney test, one-way ANOVA test and Fisher’s exact, and two-tailed Pearson correlation."}}, {"id": "method-2", "family": "group_comparison", "specific": "anova", "label": "Group comparison", "dsl_operations": ["group_summary", "welch_t", "bootstrap_mean_difference", "group_comparison_anova"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 0.8, "band": "high", "quote": "ANOVA"}}, {"id": "method-3", "family": "correlation_analysis", "specific": "pearson", "label": "Correlation analysis", "dsl_operations": ["pearson_r"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["paired_numeric_columns"], "evidence": {"score": 0.8, "band": "high", "quote": "Pearson correlation"}}], "targets": [], "gaps": []}, "capability": {"state": "complete", "requirements": [{"capability_id": "capability:computational_analysis", "label": "Computational analysis platform", "assay_families": ["meta_analysis", "machine_learning", "survival_analysis", "statistical_regression", "pca", "molecular_docking", "network_analysis", "differential_expression", "gwas"], "availability_state": "owned", "tier_unlocked": "R1", "candidate": "Automated Assays Laravel application and simulated adapters"}], "gaps": []}, "readiness": {"state": "complete", "tier": "R1", "blocking_axis": "none", "blocked_on": "nothing:replicable_now", "axes": {"method": {"tier": "R1", "expressible": true, "required_operations": ["welch_t"], "missing_operations": [], "basis": "The paper names the method it used and the corpus's method extractor maps it onto the DSL catalogue: welch_t_test."}, "capability": {"tier": "R1", "capability_id": "capability:computational_analysis", "availability_state": "owned", "basis": "The computational analysis is owned. 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Information regarding such variants must not be shared publicly following the European Union legislation described in the following document: https://publications.jrc.ec.europa.eu/repository/bitstream/JRC113479/policy_report_-_review_of_eu_national_legislation_on_genomics_-_with_identifiers_1.pdf, therefore the access to raw data that support findings of this study are available from the corresponding author upon reasonable request."], "basis": "The paper names GSE207805 in Gene Expression Omnibus (GEO) and the accession resolves in the repository. 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STUDY DESIGN SIZE DURATION: Using the CRISPR/Cas9 genome editing technique, a mouse Tcte1", "Materials and methods Animals Using the CRISPR/Cas9 genome editing technique, the mouse het", "matic representation of gRNA localization in CRISPR/Cas9 KO creation. (B) Relative expression of"], "order_basis": "paper_text_order:996", "id": "assay:1"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ion levels of corresponding genes identified in the mouse testis RNAseq experiments. Immunofluorescenc", "ed variants and the exon 3 structure deleted in the mouse knockout was also performed. MAIN RESULTS AN", "the CRISPR/Cas9 genome editing technique, a mouse Tcte1 gene knockout line was created on the", "ted on the basis of the C57Bl/6J strain. The mouse reproductive potential, semen characteristic"], "order_basis": "paper_text_order:1037", "id": "assay:2"}, {"role": "data_generation", "definition_id": "apoptosis_assay", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ene expression levels, sperm ATP, and testis apoptosis level measurements were then assessed, follo", "staining was performed to check the level of apoptosis via caspase 3 visualization on mouse testis", "mitochondrial ATP processing or linked with apoptosis or spermatogenesis. In Tcte1-/- males, the p", "beating. Casp3-positive signals (indicating apoptosis) were observed in spermatogonia only, at a s"], "order_basis": "paper_text_order:1230", "id": "assay:3"}, {"role": "data_generation", "definition_id": "rna_seq", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["llowed by semen analysis. Sequencing of RNA (RNAseq; Illumina) was done for mice testis tissues.", "ponding genes identified in the mouse testis RNAseq experiments. Immunofluorescence in situ stai", "gozoospermia, suggesting haploinsufficiency. RNA-sequencing of the testicular tissue showed the influenc", "cle and its supplementary information files. RNAseq data are available in the GEO database (http"], "order_basis": "paper_text_order:1964", "id": "assay:4"}, {"role": "data_generation", "definition_id": "ihc", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["fied in the mouse testis RNAseq experiments. Immunofluorescence in situ staining was performed to detect the", "uminescence level was measured. In addition, immunofluorescence in situ staining was performed to check the"], "order_basis": "paper_text_order:2192", "id": "assay:5"}, {"role": "data_generation", "definition_id": "qpcr", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ion of the Tcte1 gene was verified using the real-time PCR method (Fig. 2B; Supplementary Data File S2)", "T), hetero- (+/−) and homozygous mice (−/−) (Real-time PCR); **** P < 0.0001, ** P < 0.01. (C) Example"], "order_basis": "paper_text_order:6938", "id": "assay:6"}, {"role": "analysis", "definition_id": "group_comparison", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["welch_t"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["Statistical analyses included the following: normality test (D’Agostino and Pearson or Kolmogorov–Smirnoff), unpaired two-tailed t-test with Welch’s correction or Mann–Whitney test, one-way ANOVA test and Fisher’s exact, and two-tailed Pearson correlation."], "order_basis": "consumes_earlier_output", "id": "assay:7"}]}