CRISP2 Is a Regulator of Multiple Aspects of Sperm Function and Male Fertility

Lim, Shuly; Kierzek, Michelina; O’Connor, Anne E; Brenker, Christoph; Merriner, D. Jo; Okuda, Hidenobu; Volpert, Marianna; Gaikwad, Avinash; Bianco, Deborah; Potter, David; Prabhakar, Ranganathan; Strünker, Timo; O'Bryan, Moira K

doi:10.1210/en.2018-01076

Cited by 53 publications

(80 citation statements)

References 56 publications

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“…However, when fertilization in the oviduct was evaluated under more challenging conditions such as mating double mutant males with superovulated wild-type females, 27 a marked decrease in fertilization rates was observed in Crisp2 −/− .Crisp4 −/− males compared to controls or to either Crisp2 or Crisp4 single mutant males ( Figure 1B). Consistent with these results and the reported roles for CRISP2 and CRISP4 in fertilization, 7,27,28,35 sperm from Crisp2 −/− .Crisp4 −/− mice showed severe defects in their ability to fertilize cumulus-oocyte complexes in vitro ( Figure 1C) as well as in several capacitation-associated parameters such as tyrosine phosphorylation, progesterone-induced acrosome reaction and hyperactivation ( Figure 1D-F).…”

Section: Generation and Characterization Ofsupporting

confidence: 88%

“…Previous studies involving deletions of individual Crisp genes by homologous recombination showed normal fertility in mutant males despite of several defects found when sperm were tested in in vitro fertilization assays. 7,14,[26][27][28]33 These results, together with the finding that Crisp1 −/− .Crisp4 −/− mutant males exhibited a mild subfertility 7 supported the possible existence of partially redundant and compensatory mechanisms among CRISP family members. Based on these observations, in the present study, we investigated the relevance of CRISP proteins for male fertility by generating mutant mice carrying simultaneous null mutations in several Crisp genes using CRISPR/Cas9 technology.…”

Section: Discussionmentioning

confidence: 61%

“…26 Consistent with this idea, mice lacking Crisp2 were also fertile in spite of exhibiting several sperm fertilizing deficiencies under in vitro conditions. 27,28 However, when the reproductive performance of these animals was challenged by either reducing the number of sperm in their ejaculates (through unilateral vasectomy) or increasing the number of oocytes to be fertilized in vivo (using superovulated females), Crisp2 −/− males showed significantly lower fertility rates compared to their wild-type littermates. 27 These observations support the idea that both CRISP1 and CRISP2 play important roles in the fertilization process that are partially masked in each individual gene knockout mouse model by compensatory mechanisms and become evident only under more demanding conditions that better reflect the natural environment.…”

Section: Introductionmentioning

confidence: 99%

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Functional redundancy and compensation: Deletion of multiple murineCrispgenes reveals their essential role for male fertility

et al. 2020

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Mammalian Cysteine-RIch Secretory Protein (CRISP) family includes four members present in sperm and reported to regulate Ca 2+ channels and fertilization. Based on our previous observations using single knockouts models and suggesting the existence of functional compensation among CRISP proteins, we investigated their relevance for male fertility by generating multiple Crisp gene mutants by CRISPR/ Cas9 technology. Whereas targeting of Crisp1 and Crisp3 yielded subfertile males with early embryo developmental defects, the same deletion in zygotes from fertile Crisp2 −/− .Crisp4 −/− mice led to the generation of both triple and quadruple knockout mice exhibiting a complete or severe disruption of male fertility due to a combination of sperm transport, fertilization, and embryo developmental defects linked to intracellular Ca 2+ dysregulation. These observations reveal that CRISP proteins are essential for male fertility and organize in functional modules that contribute distinctly to fertility success, bringing insights into the mechanisms underlying functional redundancy/compensation in protein families and emphasizing the importance of generating multiple and not just single knockout which might be masking the true functional relevance of family genes.

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Section: Generation and Characterization Ofsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 61%

Section: Introductionmentioning

confidence: 99%

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Functional redundancy and compensation: Deletion of multiple murineCrispgenes reveals their essential role for male fertility

et al. 2020

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“…The mouse knockout line were maintained on a C57/BL6N background. Sperm were collected from cauda epididymides and vas deferens using the back-flushing method [33] in modified TYH medium…”

Section: A Sperm Sample Preparationmentioning

confidence: 99%

Flagellar energetics from high-resolution imaging of beating patterns in tethered mouse sperm

Nandagiri

Gaikwad

Potter

et al. 2020

Preprint

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While much is known about the microstructure of sperm flagella, the mechanisms behind the generation of flagellar beating patterns by the axoneme are still not fully understood. We demonstrate a technique for investigating the energetics of flagella or cilia. We record the planar beating of tethered wildtype and Crisp2-knockout mouse sperm at high-speed and high-resolution and extract centerlines using digital image processing techniques. We accurately reconstruct beating waveforms using a Chebyshev-polynomial based Proper Orthogonal Decomposition of the centerline tangent-angle profiles. External hydrodynamic forces and the internal resistance from the passive flagellar material are calculated from the observed kinematics of the beating patterns using a Soft, Internally-Driven Kirchhoff-Rod (SIDKR) model. Energy conservation is employed to further compute the flagellar energetics. We thus obtain the distribution of mechanical power exerted by the dynein motors without any further assumptions about mechanisms regulating axonemal function. We find that, in both the mouse genotypes studied, a large proportion of the mechanical power exerted by the dynein motors is dissipated internally, within the passive structures of the flagellum and by the motors themselves. This internal dissipation is considerably greater than the hydrodynamic dissipation in the aqueous medium outside. The net power input from the dynein motors in sperm from Crisp2-knockout mice is significantly smaller than in corresponding wildtype samples. The reduced power is correlated with slower beating and smaller amplitudes. These measurements of flagellar energetics indicate that the ion-channel regulating cysteine-rich secretory proteins (CRISPs) may also be involved in regulating mammalian sperm motility.

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“…Moreover, considering the high intracellular calcium levels observed in CRISP2 mutant spermatozoa (Brukman et al ., ), the defects in hyperactivation might be linked to the reported ability of CRISP2 to regulate calcium channels (Gibbs et al ., ). Interestingly, motility and fertility disorders were also reported in patients with aberrant expression of CRISP2 (Du et al ., ; Zhou et al ., ) as well as in a recently reported KO for CRISP2 (Lim et al ., ).…”

Section: Cysteine Rich Secretory Proteinmentioning

confidence: 97%

Relevance of CRISP proteins for epididymal physiology, fertilization, and fertility

et al. 2019

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Background: The molecular mechanisms involved in the acquisition of mammalian sperm fertilizing ability are still poorly understood, reflecting the complexity of this process. Objectives: In this review, we describe the role of Cysteine RIch Secretory Proteins (CRISP1-4) in different steps of the sperm journey to the egg as well as their relevance for fertilization and fertility. Materials and Methods: We analyze bibliography reporting the phenotypes of CRISP KO mice models and combine this search with recent findings from our team. Results: Generation of individual KO for CRISP proteins reveals they are key mediators in different stages of the fertilization process. However, in spite of their important functional roles, KO males for each of these proteins remain fertile, supporting the existence of compensatory mechanisms between homologous CRISP family members. The development of mice lacking epididymal CRISP1 and CRISP4 simultaneously (DKO) revealed that mutant males exhibit an impaired fertility due to deficiencies in the sperm ability to fertilize the eggs in vivo, consistent with the proposed roles of the two proteins in fertilization. Interestingly, DKO males show clear defects in both epididymal epithelium differentiation and luminal acidification known to be critical for sperm maturation and storage. Whereas in most of the cases, these epithelium defects seem to specifically affect the sperm fertilizing ability, some animals exhibit a disruption of the characteristic immune tolerance of the organ with clear signs of inflammation and sperm viability defects. Discussion and Conclusion: Altogether, these observations confirm the relevance of CRISP proteins for male fertility and contribute to a better understanding of the fine-tuning mechanisms underlying sperm maturation and immune tolerance within the epididymis. Moreover, considering the existence of a human epididymal protein functionally equivalent to rodent CRISP1 and CRISP4, DKO mice may represent an excellent model for studying human epididymal physiology and pathology.

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CRISP2 Is a Regulator of Multiple Aspects of Sperm Function and Male Fertility

Cited by 53 publications

References 56 publications

Functional redundancy and compensation: Deletion of multiple murineCrispgenes reveals their essential role for male fertility

Functional redundancy and compensation: Deletion of multiple murineCrispgenes reveals their essential role for male fertility

Flagellar energetics from high-resolution imaging of beating patterns in tethered mouse sperm

Relevance of CRISP proteins for epididymal physiology, fertilization, and fertility

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