Distinct mechanisms underlie sperm-induced and proteaseinduced oolemma block to sperm penetration

Komorowski, Sebastian; Szczepańska, Katarzyna; Maleszewski, Marek

doi:10.1387/18

Cited by 11 publications

(12 citation statements)

References 19 publications

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“…CD9 has also been reported to be an important factor for sperm-oolemma fusion in mouse, bovine and porcine oocytes [56−58]. Other research has indicated that mouse oocytes incubated with pronase to remove the ZP lose all their CD9 from the oolemma [59]. Our present findings support this possibility.…”

Section: Discussionsupporting

confidence: 87%

Evaluation of Zona Pellucida Function for Sperm Penetration During <i>In Vitro</i> Fertilization in Pigs

et al. 2013

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In porcine oocytes, the function of the zona pellucida (ZP) with regard to sperm penetration or prevention of polyspermy is not well understood. In the present study, we investigated the effects of the ZP on sperm penetration during in vitro fertilization (IVF). We collected in vitro-matured oocytes with a first polar body (ZP+ oocytes). Some of them were freed from the ZP (ZP− oocytes) by two treatments (pronase and mechanical pipetting), and the effects of these treatments on sperm penetration parameters (sperm penetration rate and numbers of penetrated sperm per oocyte) were evaluated. There was no evident difference in the parameters between the two groups. Secondly, we compared the sperm penetration parameters of ZP+ and ZP− oocytes using frozen-thawed epididymal spermatozoa from four boars. Sperm penetration into ZP+ oocytes was found to be accelerated relative to ZP− oocytes. Thirdly, we evaluated the sperm penetration of ZP+ and ZP− oocytes at 1−10 h after IVF (3 h gamete co-incubation). The proportions of oocytes penetrated by sperm increased significantly with time in both groups; however, the number of penetrated sperm per oocyte did not increase in ZP− oocytes. Finally, we performed IVF using ZP− oocytes divided into control (3 h) and prolonged gamete co-incubation (5 h) groups. Greater numbers of sperm penetrated in the 5 h group than in the control group. These results suggest that the ZP and oolemma are not competent factors for prevention of polyspermy in our present porcine IVF system. However, it appears that ZP removal is one of the possibilities for reducing polyspermic penetration in vitro in pigs.

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Section: Discussionsupporting

confidence: 87%

Evaluation of Zona Pellucida Function for Sperm Penetration During <i>In Vitro</i> Fertilization in Pigs

et al. 2013

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“…However, we previously observed (using in situ immunofluorescence and western blot techniques) that in mouse zygotes the loss of oolemma fusibility with spermatozoa is not related to the decrease of CD9 protein in the oolemma (Komorowski et al, 2003). We found that the level of CD9 in zygotes was almost two times lower than in unfertilized oocytes.…”

Section: Discussionmentioning

confidence: 51%

Decrease in CD9 content and reorganization of microvilli may contribute to the oolemma block to sperm penetration during fertilization of mouse oocyte

Żyłkiewicz

Nowakowska

Maleszewski

2009

Zygote

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Tetraspanin CD9 is the only protein of the oocyte membrane (oolemma) known to be required for the fusion of gametes during fertilization in the mouse. Using electron microscopy and immunostaining we examined the differences in localization of CD9 between ovulated oocytes, zygotes and parthenogenetically activated eggs (parthenogenotes). Changes in ultrastructure of oolemma, which take place in oocytes after fertilization or artificial activation, were also assessed. We demonstrated that after fertilization the level of CD9 present on microvilli of zygote was two times lower than its level on the oolemma of the oocyte. In addition, we showed that the distribution of microvilli is less uniform in the zygotes than in the unfertilized oocytes. We propose that the changes of microvilli distribution and their CD9 content are responsible for the development of the oocyte membrane block to sperm penetration.

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“…Perhaps the possible deleterious enzymes contained in the crude collagenase are at a very low concentration or have only mild activity that can be inhibited by the albumin in the medium. Thus, it is suggested that collagenase treatment is safer than the other enzymes used for zona removal, such as a-chymotrypsin, trypsin, and pronase, which readily damage the oolemma [4,[10][11][12][13].…”

Section: Discussionmentioning

confidence: 99%

“…Enzymatic methods often use a-chymotrypsin, pronase, or trypsin [7][8][9]. Enzymatic methods using such trypsin-like enzymes are able to treat large numbers of eggs at once, but the excess treatment greatly reduces the sperm-egg fusion rate [4,[10][11][12]. There are two acidic buffer methods; one is to immerse the eggs in the acidified Tyrode's solution, and the other is to slowly blow acidified Tyrode's solution on the ZP in neutral buffer.…”

Section: Introductionmentioning

confidence: 99%

One‐step collagenase method for zona pellucida removal in unfertilized eggs: easy and gentle method for large‐scale preparation

Yamatoya

Ito

Araki

et al. 2011

Reprod Medicine & Biology

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Distinct mechanisms underlie sperm-induced and proteaseinduced oolemma block to sperm penetration

Cited by 11 publications

References 19 publications

Evaluation of Zona Pellucida Function for Sperm Penetration During <i>In Vitro</i> Fertilization in Pigs

Evaluation of Zona Pellucida Function for Sperm Penetration During <i>In Vitro</i> Fertilization in Pigs

Decrease in CD9 content and reorganization of microvilli may contribute to the oolemma block to sperm penetration during fertilization of mouse oocyte

One‐step collagenase method for zona pellucida removal in unfertilized eggs: easy and gentle method for large‐scale preparation

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