Variability of the Response to Pentoxifylline in Vitro in Infertile Normozoospermic and Asthenozoospermic Patients

Mathieu, Chantal; Écochard, René; Lornage, Jacqueline; Cordonier, H.; Guérin, J. F.

doi:10.3109/01485019408987801

Cited by 5 publications

(4 citation statements)

References 31 publications

(12 reference statements)

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“…In addition to the inter‐donor variability observed for in vitro longevity, a large variation was also observed in the proportion of Y‐chromosome‐bearing spermatozoa bound to hemizona at 24 and 48 h. These variations are not unexpected since inter‐donor variation has been reported for in vitro characteristics in many studies utilizing semen from both fertile donors and infertile patients (Perreault & Rogers, 1982; Rogers et al ., 1983; Mathieu et al ., 1994). In their report on inter‐donor variations in capacitation patterns, Perreault & Rogers (1982) stressed careful consideration of this variability when assessing fertilizing potential.…”

Section: Discussionmentioning

confidence: 99%

Differential binding of X- and Y-chromosome-bearing human spermatozoa to zona pellucidain vitro

2001

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The sex of human offspring has been associated with the day in the mother's menstrual cycle on which insemination occurs, with male zygotes being formed earlier in the fertile period than female zygotes. Using an in vitro environment designed to mimic the in vivo milieu, we tested the hypothesis that Y-chromosome-bearing spermatozoa survive functionally longer than X-chromosome-bearing spermatozoa, and that this differential functional survival is a contributing factor to the in vivo phenomenon. Donor semen was processed by swim-up and incubated at 37 degrees C in culture medium for 0, 24 and 48 h, with human zona pellucida (hemizona, HZ) being used to select functional spermatozoa. A second set of in vitro storage conditions, 4 degrees C in test-yolk refrigeration buffer, was used to determine whether changing the incubation conditions alters the process. The sex chromosome of the spermatozoa was determined using fluorescent in situ hybridization (FISH). For spermatozoa incubated at 37 degrees C, the percentage of functional (HZ bound) Y-bearing spermatozoa was significantly increased (P < 0.05) at 48 h (55.4 + 2.9%) but not at 0 h (50.5 + 0.7%) or 24 h (52.8 + 3.1%) compared to swim-up spermatozoa (50.6 + 0.3%). No difference in the percentage of functional Y-spermatozoa was observed at any time-point with storage at 4 degrees C in refrigeration buffer. Thus, we demonstrated that significantly more Y-bearing spermatozoa were capable of zona binding than X-bearing spermatozoa at 48 h at 37 degrees C incubation, with an observed Y : X ratio of 1.15 for these zona-bound spermatozoa compared to 1.02 for post-swim-up spermatozoa. We conclude that a differential functional survival appears to exist between X-bearing and Y-bearing spermatozoa, with the latter exhibiting a longer functional survival under in vitro conditions.

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Section: Discussionmentioning

confidence: 99%

Differential binding of X- and Y-chromosome-bearing human spermatozoa to zona pellucidain vitro

2001

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“…IBMX, for example, is used at concentrations from 30 μM to 1 mM ( Lefievre et al , 2000 ; Pons-Rejraji et al , 2011 ; Tardif et al , 2014 ; Marechal et al , 2017 ). Similarly, pentoxifylline has been used at 3–4 mM ( Tesarik et al , 1992 ; Burger et al , 2000 ; Patrizio et al , 2000 ; Terriou et al , 2000 ), although conflicting reports have found no improvement in human sperm motility at the same concentrations ( Mathieu et al , 1994 ; Tournaye et al , 1994 ) and higher concentrations of 10 mM have been used to examine its effects on spermatozoa DNA damage ( Banihani et al , 2018 ). Other such PDE inhibiting compounds included Milrinone, a PDE3 inhibitor shown to effect human spermatozoa motility at 50 μM ( Lefièvre et al , 2002 ), and rolipram, a PDE4 inhibitor with effects at 10 µM ( Marechal et al , 2017 ).…”

Section: Discussionmentioning

confidence: 99%

Compounds enhancing human sperm motility identified using a high-throughput phenotypic screening platform

et al. 2022

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STUDY QUESTION Can a high-throughput screening (HTS) platform facilitate male fertility drug discovery? SUMMARY ANSWER An HTS platform identified a large number of compounds that enhanced sperm motility. WHAT IS KNOWN ALREADY Several efforts to find small molecules modulating sperm function have been performed but none have used high-throughput technology. STUDY DESIGN, SIZE, DURATION Healthy donor semen samples were used and samples were pooled (3–5 donors per pool). Primary screening was performed singly; dose–response screening was performed in duplicate (using independent donor pools). PARTICIPANTS/MATERIALS, SETTING, METHODS Spermatozoa isolated from healthy donors were prepared by density gradient centrifugation and incubated in 384-well plates with compounds (6.25 μM) to identify those compounds with enhancing effects on motility. Approximately 17 000 compounds from the libraries, ReFRAME, Prestwick, Tocris, LOPAC, CLOUD and MMV Pathogen Box, were screened. Dose–response experiments of screening hits were performed to confirm the enhancing effect on sperm motility. Experiments were performed in a university setting. MAIN RESULTS AND THE ROLE OF CHANCE From our primary single concentration screening, 105 compounds elicited an enhancing effect on sperm motility compared to dimethylsulphoxide-treated wells. Confirmed enhancing compounds were grouped based on their annotated targets/target classes. A major target class, phosphodiesterase inhibitors, were identified, in particular PDE10A inhibitors as well as number of compounds not previously known to enhance human sperm motility, such as those related to GABA signalling. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION Although this approach provides data about the activity of the compound, it is only a starting point. For example, further substantive experiments are necessary to provide a more comprehensive picture of each compound’s activity, the effect on the kinetics of the cell populations and subpopulations, and their potential mechanisms of action. Compounds have been tested with prepared donor spermatozoa, incubated under non-capacitating conditions, and only incubated with compounds for a relatively short period of time. Therefore, the effect of compounds under different conditions, for example in whole semen, for longer incubation times, or using samples from patient groups, may be different and require further study. All experiments were performed in vitro. WIDER IMPLICATIONS OF THE FINDINGS This phenotypic screening assay identified a large number of compounds that increased sperm motility. In addition to furthering our understanding of human sperm function, for example identifying new avenues for discovery, we highlight potential compounds as promising start-point for a medicinal chemistry programme for potential enhancement of male fertility. Moreover, with disclosure of the results of screening, we present a substantial resource to inform further work in the field. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the Bill and Melinda Gates Foundation, Scottish Funding Council and Scottish Universities Life Science Alliance. C.L.R.B. is Editor for RBMO. C.L.R.B. receives funding from Chief Scientists Office (Scotland), ESHRE and Genus PLC, consulting fees from Exscientia and lecture fees from Cooper Surgical and Ferring. S.M.d.S. is an Associate Editor of Human Reproduction, and an Associate Editor of Reproduction and Fertility. S.M.d.S. receives funding from Cooper Surgical and British Dietetic Society. No other authors declared a COI.

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“…IBMX, for example, is used at concentrations from 30 μM to 1 mM (Lefievre et al, 2000; Marechal et al, 2017; Pons-Rejraji et al, 2011; Tardif et al, 2014). Similarly, pentoxifylline has been used at 3 to 4 mM (Burger et al, 2000; Patrizio et al, 2000; Terriou et al, 2000; Tesarik et al, 1992) although conflicting reports have found no improvement in human sperm motility at the same concentrations (Mathieu et al, 1994; Tournaye et al, 1994) and higher concentrations of 10mM have been used to examine its effects on spermatozoa DNA damage (Banihani et al, 2018). Other such PDE inhibiting compounds included Milrinone, a PDE3 inhibitor shown to effect human spermatozoa motility at 50 μM (Lefiévre et al, 2002), and rolipram, a PDE4 inhibitor with effects at 10 µM (Marechal et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

Compounds enhancing human sperm motility identified using a high-throughput phenotypic screening platform

Gruber

Johnston

Norcross

et al. 2021

Preprint

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Study question: Can a high-throughput screening platform facilitate male fertility drug discovery? Summary answer: A high-throughput screening platform identified a large number of compounds that enhanced sperm motility. What is known already: Several efforts to find small molecules modulating sperm function have been performed but not using high-throughput technology. Study design, size, duration: Healthy donor semen samples were used and samples were pooled (3-5 donors per pool). Primary screening was performed in singlicate; dose-response screening was performed in duplicate (independent donor pools). Participants/materials, setting, methods: Spermatozoa isolated from healthy donors were prepared by density gradient centrifugation and incubated in 384-well plates with compounds (6.25 uM) to identify those compounds with enhancing effects on motility. A total of ~17,000 compounds from the following libraries: ReFRAME, Prestwick, Tocris, LOPAC, CLOUD and MMV Pathogen Box were screened. Dose response experiments of screening hits were performed to confirm the enhancing effect on sperm motility. Experiments were performed in a University setting. Main results and the role of chance: From our primary single concentration screening, 105 compounds elicited an enhancing effect on sperm motility compared to DMSO treated wells. Confirmed enhancing compounds were grouped based on their annotated targets/target classes. A major target class, phosphodiesterase inhibitors, were identified in particular PDE10A inhibitors as well as number of compounds not previously identified/known to enhance human sperm motility such as those related to GABA signaling. Limitations, reasons for caution: Compounds have been tested with prepared donor spermatozoa and only incubated for a short period of time. Therefore, the effect of compounds on whole semen or with longer incubation time may be different. All experiments were performed in vitro. Wider implications of the findings: This phenotypic screening assay identified a large number of compounds that increased sperm motility. In addition to furthering our understanding of human sperm function, for example identifying new avenues for discovery, we highlight potential inhibitors as promising start-point for a medicinal chemistry programme for potential enhancement of male infertility. Moreover, with disclosure of the results of screening we present a substantial resource to inform further work in the field.

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Variability of the Response to Pentoxifylline in Vitro in Infertile Normozoospermic and Asthenozoospermic Patients

Cited by 5 publications

References 31 publications

Differential binding of X- and Y-chromosome-bearing human spermatozoa to zona pellucidain vitro

Differential binding of X- and Y-chromosome-bearing human spermatozoa to zona pellucidain vitro

Compounds enhancing human sperm motility identified using a high-throughput phenotypic screening platform

Compounds enhancing human sperm motility identified using a high-throughput phenotypic screening platform

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