Dissect Assembly Rules of SPET12 Complex in the Mammalian Sperm

NCT04159402 | Completed

• 1 other identifier: A-ER-102-438
• Study Type: observational
• Target: 183
• Locations: 0 countries
• Sites: N/A

Brief Summary

The sperm of the KO mouse shothe investigatorsd severe defects of all found compartments: acrosome, nucleus, midpiece and tail. The investigators also found SEPT12 co-localizes and interact with SEPT1, 2, 4, 6, 7, 10, 11, and 14 in sperm. Interestingly, some of these septins form filaments with SEPT12 in cells. Based on these findings, it is hypothesized that (1) The core complex of SEPT12 filament consists of SEPT12-7-6-2-2-6-7-12; (2) Septin12 mutations, genetic variants, as the investigatorsll as haploinsufficiency disrupt SEPT12 filament and macro-complex; (3) Other SEPT's (e.g. SETP4, SEPT14) are also involved in the functionality of SEPT12; and (4) SEPT12 macro-complex is critical for compartment formation during terminal differentiation of male germ cells. The proposed study is designed to confirm the above hypotheses, to deconstruct mammalian SEPT12 complex, and to elucidate the functional significance of Septin12 mutations. In the proposed study, the investigators will use different methods, including proteomics, immunofluorescence assay, co- immunoprecipitation, pull-down assay, protein domain mapping, and protein complex fractionation to test the above hypothesis. The investigators have created a mouse carrying an important Septin12 mutation. In the proposed study, the investigators plan to knock out Septin14 in the mouse. SEPT14 interacts with SEPT12 and is also predominantly expressed in sperm. The mouse models and research tools could be used to explore the role of septins during spermiogenesis, to deconstruct the SEPT12 complex in the mammalian sperm, and to elucidate the functional significance of the Septin 12 mutations. Our findings may provide novel insight into the pathways human spermatogenesis, and has the potential to lead to development of therapeutic models for male infertility, and design of male contraceptives.

Conditions

Spermatozoa; Fertility

Outcome Measures

Primary Outcomes (1)

• Losses of SEPT12, SEPT7, SEPT6, SEPT2 and SEPT4 from the sperm annulus of a SEPT12D197N patient.

  Immunofluorescence staining of normal sperm in a fertile control showed signals for SEPT12 . SEPT7, SEPT6, SEPT2 and SEPT4 (red) at the annulus. Also shown are merged fluorescence staining images with the additional staining of the sperm nuclei, as well as merged fluorescence staining images with merged brightfield images (BF merged). Blue color, DAPI stain; BF, brightfield only. Scale bar: 10 µm.

  1 day

Eligibility Criteria

• Age: 20 Years - 50 Years
• Sex: male
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Probability Sample

Study Population

The participants were clinically diagnosed by semen analysis and further collected the normal human sperm.

You may qualify if:

• All participants signed a written informed consent form.
• Semen samples were obtained by masturbation after 3-5 days of sexual abstinence.

You may not qualify if:

• ．None

Sponsors & Collaborators

• National Cheng-Kung University Hospital: lead

Biospecimen

Retention: SAMPLES WITH DNA

Sperm

Study Officials

• Pao-Lin Kuo, MD

  Department of Obstetrics and Gynecology, National Cheng Kung University Hospital

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: October 28, 2019
• First Posted: November 12, 2019
• Study Start: August 1, 2014
• Primary Completion: July 1, 2017
• Study Completion: July 1, 2017
• Last Updated: October 4, 2022

Record last verified: 2019-11