Capacitation-associated Protein Tyrosine Phosphorylation As a Possible Biomarker of Sperm Selection

NCT04962100 | Completed

• 1 other identifier: 1608-SEV-065-CG
• Study Type: interventional
• Target: 335
• Locations: 1 country
• Sites: 1

Brief Summary

Sperm undergo complex selection processes and physiological changes as they move through the female reproductive tract. Ejaculated-sperm must undergo a set of molecular and biochemical changes globally named as capacitation in order to acquire the ability to fertilize the oocyte. These changes include post-translational modifications of sperm proteins, with phosphorylation of tyrosine residues being one of the most outstanding characteristics of the capacitation process. In the laboratory, the capacitation process is recreated artificially before performing artificial insemination or in vitro fertilization treatments. The sample is then incubated until it is used in the treatment. Reproductive success rates can be affected by differences in incubation times and levels of capacitation of the sample. In this study, the investigators intend to study the capacitation state of the sample by measuring the levels of phosphorylation of the tyrosine residues of the proteins contained in the sperm that have already been subjected to the capacitation process in vitro.

Conditions

Sperm Capacitation

Keywords

Capacitation; Tyrosine phosphorylation; Flow cytometry; Biomarker

Outcome Measures

Primary Outcomes (1)

• To determine the state of sperm capacitation by assessing the phosphorylation of tyrosine residues

  Flow cytometry analysis will be carried out to evaluate the levels of phosphorylation of tyrosine residues in sperm proteins of seminal samples capacitated in vitro in order to determine their capacitation statuscytometry of seminal samples capacitated in vitro in order to determine their capacitation status

  1 day

Study Arms (1)

Study population

EXPERIMENTAL

Male patients and donors who provide a sample of fresh ejaculated semen will be the population of this study, as well as female patients undergoing artificial insemination with their partner's semen or frozen donor semen.

Other: Analysis of capacitation-associated tyrosine phosphorylation in human sperm by flow cytometry

Interventions

Analysis of capacitation-associated tyrosine phosphorylation in human sperm by flow cytometry OTHER

After the assessment of the macroscopic and microscopic characteristics of the sample, we will wash the sample and process it using three layers of density gradients. In the case of samples for diagnostic analysis, the remaining volume will be incubated in at 37ºC, and 0.1-0.2 ml aliquots will be taken from the end of the processing at t0 (just before introducing the sample in the incubator), at t1 (after one hour of incubation) and at t3 (after 3 hours of incubation). Additionally, a sample will be taken after washing, prior to processing the sample with the density gradients. In the case of samples processed for insemination, an additional aliquot will be taken at the exact moment in which the insemination is carried out and the elapsed incubation time will be noted. The evaluation of the capacitation will be carried out by analyzing the state of phosphorylation of tyrosines by flow cytometry.

Study population

Eligibility Criteria

• Age: 18 Years+
• Sex: male
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• To have signed the written informed consent before starting any procedure related to the study.
• Sexual abstinence (2 to 5 days).
• To be a male patient who delivers a fresh seminal sample for diagnosis or artificial insemination treatment.
• To be a male donor who delivers a fresh seminal sample for donation or for previous tests to include him in the donation programme.
• To be a female patient undergoing artificial insemination.

You may not qualify if:

• Use of sperm of testicular or epididymal origin.
• Cryptozoospermia or oligozoospermia with sperm concentration \<1 mill/ml.
• Ejaculates obtained with more than 5 days of sexual abstinence.
• Frozen semen samples for artificial insemination cycles with partner's semen.
• Female patients who present uterine malformations that compromise the viability of the pregnancy (intramural or submucosal fibroids \> 3 cm, polyps, adenomyosis or congenital or acquired malformations). Female patients with hydrosalpinx.
• Patients who have suffered two or more previous abortions (repeated abortions).
• Patients with a body mass index greater than 30 Kg / m2.

Sponsors & Collaborators

• Vida Recoletas Sevilla: lead

Study Sites (1)

IVI RMA Sevilla

Seville, Sevilla, 41092, Spain

Location

Related Publications (13)

• Said TM, Land JA. Effects of advanced selection methods on sperm quality and ART outcome: a systematic review. Hum Reprod Update. 2011 Nov-Dec;17(6):719-33. doi: 10.1093/humupd/dmr032. Epub 2011 Aug 25.

  PMID: 21873262 BACKGROUND

• Aitken RJ, Nixon B. Sperm capacitation: a distant landscape glimpsed but unexplored. Mol Hum Reprod. 2013 Dec;19(12):785-93. doi: 10.1093/molehr/gat067. Epub 2013 Sep 26.

  PMID: 24071444 BACKGROUND

• Bianchi E, Doe B, Goulding D, Wright GJ. Juno is the egg Izumo receptor and is essential for mammalian fertilization. Nature. 2014 Apr 24;508(7497):483-7. doi: 10.1038/nature13203. Epub 2014 Apr 16.

  PMID: 24739963 BACKGROUND

• Henkel R. Sperm preparation: state-of-the-art--physiological aspects and application of advanced sperm preparation methods. Asian J Androl. 2012 Mar;14(2):260-9. doi: 10.1038/aja.2011.133. Epub 2011 Dec 5.

  PMID: 22138904 BACKGROUND

• Visconti PE, Bailey JL, Moore GD, Pan D, Olds-Clarke P, Kopf GS. Capacitation of mouse spermatozoa. I. Correlation between the capacitation state and protein tyrosine phosphorylation. Development. 1995 Apr;121(4):1129-37. doi: 10.1242/dev.121.4.1129.

  PMID: 7743926 BACKGROUND

• Visconti PE. Understanding the molecular basis of sperm capacitation through kinase design. Proc Natl Acad Sci U S A. 2009 Jan 20;106(3):667-8. doi: 10.1073/pnas.0811895106. Epub 2009 Jan 14. No abstract available.

  PMID: 19144927 BACKGROUND

• Liu DY, Clarke GN, Baker HW. Tyrosine phosphorylation on capacitated human sperm tail detected by immunofluorescence correlates strongly with sperm-zona pellucida (ZP) binding but not with the ZP-induced acrosome reaction. Hum Reprod. 2006 Apr;21(4):1002-8. doi: 10.1093/humrep/dei435. Epub 2006 Jan 20.

  PMID: 16428332 BACKGROUND

• Kumaresan A, Siqueira AP, Hossain MS, Johannisson A, Eriksson I, Wallgren M, Bergqvist AS. Quantification of kinetic changes in protein tyrosine phosphorylation and cytosolic Ca(2)(+) concentration in boar spermatozoa during cryopreservation. Reprod Fertil Dev. 2012;24(4):531-42. doi: 10.1071/RD11074.

  PMID: 22541541 BACKGROUND

• Escoffier J, Navarrete F, Haddad D, Santi CM, Darszon A, Visconti PE. Flow cytometry analysis reveals that only a subpopulation of mouse sperm undergoes hyperpolarization during capacitation. Biol Reprod. 2015 May;92(5):121. doi: 10.1095/biolreprod.114.127266. Epub 2015 Apr 8.

  PMID: 25855261 BACKGROUND

• Naresh S, Atreja SK. The protein tyrosine phosphorylation during in vitro capacitation and cryopreservation of mammalian spermatozoa. Cryobiology. 2015 Jun;70(3):211-6. doi: 10.1016/j.cryobiol.2015.03.008. Epub 2015 Mar 28.

  PMID: 25828199 BACKGROUND

• Mansour RT, Serour MG, Abbas AM, Kamal A, Tawab NA, Aboulghar MA, Serour GI. The impact of spermatozoa preincubation time and spontaneous acrosome reaction in intracytoplasmic sperm injection: a controlled randomized study. Fertil Steril. 2008 Sep;90(3):584-91. doi: 10.1016/j.fertnstert.2006.11.176. Epub 2008 Mar 4.

  PMID: 18295761 BACKGROUND

• Zhang XD, Chen MY, Gao Y, Han W, Liu DY, Huang GN. The effects of different sperm preparation methods and incubation time on the sperm DNA fragmentation. Hum Fertil (Camb). 2011 Sep;14(3):187-91. doi: 10.3109/14647273.2011.604817. Epub 2011 Aug 23.

  PMID: 21859363 BACKGROUND

• Marin-Briggiler CI, Tezon JG, Miranda PV, Vazquez-Levin MH. Effect of incubating human sperm at room temperature on capacitation-related events. Fertil Steril. 2002 Feb;77(2):252-9. doi: 10.1016/s0015-0282(01)02982-x.

  PMID: 11821080 BACKGROUND

Study Officials

• Cristina González Ravina, PhD

  IVI RMA Sevilla

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NA
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: SINGLE GROUP
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: June 29, 2021
• First Posted: July 14, 2021
• Study Start: June 29, 2021
• Primary Completion: September 30, 2023
• Study Completion: September 30, 2023
• Last Updated: October 8, 2024

Record last verified: 2024-10

Data Sharing

• IPD Sharing: Will share
• Shared Documents: CSR
• Time Frame: The IPD and any additional supporting information will become available after the analysis of the data and publication of the study results.
• Access Criteria: Scientific article

Locations

ES (1)