Inadequate function of sterile tw5/tw32 spermatozoa overcome by intracytoplasmic sperm injection

Kuretake, Shoji; Maleszewski, Marek; Tokumasu, Ako; Fujimoto, Hirokazu; Yanagimachi, Ryuzo

doi:10.1002/(sici)1098-2795(199606)44:2<230::aid-mrd12>3.3.co;2-j

Cited by 13 publications

(12 citation statements)

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“…Moreover, when the gene responsible for a mutation is not known (i.e., when genotyping is not possible) and the differences in phenotype of homo-and heterozygous mice are not definite, recognizing these mice becomes problematic. The ICSI allows male infertility to be overcome, and spermatozoa that are unable to fertilize oocytes in vivo or in vitro are able to do so when injected into oocytes from normal mice, as has been shown in the present study and elsewhere [21,[27][28][29]. The azh/azh females are considered to be fertile [20], and they reproduced when mated with both wild-type and heterozygous males (present study).…”

Section: Discussionmentioning

confidence: 99%

Intracytoplasmic Sperm Injection Effects in Infertile azh Mutant Mice1

Ward

2005

Biology of Reproduction

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Several reports in the literature describe men with infertility resulting from abnormal sperm head shape or decapitation defects of their spermatozoa. These defects are similar to those shown for the spermatozoa from azh (abnormal spermatozoon head shape) mice. The present study examines the efficiency and effects of intracytoplasmic sperm injection (ICSI) in successive generations of azh mice generated with this method. Three successive generations of azh mice were produced with ICSI. In all three ICSI series, more than 80% of 2-cell embryos were obtained, and more than 35% of embryos transferred gave rise to normal live offspring. In addition, ICSI was used to cross homozygous azh/azh males with homozygous azh/azh females, and live offspring were obtained. The ICSI-derived males were tested for their fecundity and abnormalities of sperm morphology. Spermatozoa from ICSI-derived azh/+ males did not show any impairment of fecundity in in vitro fertilization. These spermatozoa successfully fertilized oocytes from both C57BL/6 and B6D2F1 females, with fertilization rates ranging from 70%- 92%. The proportion of morphologically normal spermatozoa was similar in azh/+ males from three successive generations of ICSI (57.8%, 54.8%, and 49.0%, respectively), and no differences were noted when comparing ICSI-derived males with males derived by mating (57.6%) and with wild-type controls (61.6%). Detailed analysis differentiating between specific types of anomalies of sperm morphology did not reveal significant differences among the examined groups. The results of the present study demonstrate that ICSI does not enhance the azh mutation phenotype in the offspring and brings no risks when applied continuously. Moreover, serial (successive generations) ICSI is highly efficient in maintaining valuable mice with fertility problems.

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Section: Discussionmentioning

confidence: 99%

Intracytoplasmic Sperm Injection Effects in Infertile azh Mutant Mice1

Ward

2005

Biology of Reproduction

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“…Sperm cryopreservation is more cost effective and less labor intensive than embryo freezing. Furthermore, with the advent of intracytoplasmic sperm injection (ICSI) many breeding problems exhibited by defective male reproductive function in mutant and transgenic lines, e.g., low sperm motility or concentration and/or abnormal sperm morphology, can be overcome [2][3][4][5].…”

Section: Introductionmentioning

confidence: 99%

Long-Term Preservation of Mouse Spermatozoa after Freeze-Drying and Freezing Without Cryoprotection1

Ward

Kaneko

Kusakabe

et al. 2003

Biology of Reproduction

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133

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The widespread production of mice with transgenes, disrupted genes and mutant genes, has strained the resources available for maintaining these mouse lines as live populations, and dependable methods for gamete and embryo preservation in these lines are needed. Here we report the results of intracytoplasmic sperm injection (ICSI) with spermatozoa freeze-dried or frozen without a cryoprotectant after storage for periods up to 1.5 years. Freeze-dried samples were stored at 4 degrees C. Samples frozen without cryoprotection were maintained at -196 degrees C. After storage, spermatozoa were injected into the oocytes by ICSI. Zygotic chromosomes and fetal development at Day 15 of gestation were examined after 0, 1, 3, 6, 9, and 12 mo of sperm storage. When fresh spermatozoa were used for ICSI, 96% of resultant zygotes contained normal chromosomes, and 58% of two-cell embryos transferred developed to normal viable fetuses. Similar results were obtained when spermatozoa were frozen without cryoprotection and then used for ICSI (87% and 45%, respectively; P > 0.05) and after 12 mo of sperm storage (mean of six endpoints examined: 87% and 52%, respectively; P > 0.05). Freeze-drying decreased the proportion of zygotes with normal karyoplates (75% vs. 96%; P < 0.001) and the proportion of embryos that developed into fetuses (35% vs. 58%; P < 0.001), but similar to freezing, there was no further deterioration during 12 mo of storage (mean of six endpoints examined: 68% and 34%, respectively; P > 0.05). Live offspring were obtained from both freeze-dried and frozen spermatozoa after storage for 1.5 yr. The results indicate that 1) the freeze-drying procedure itself causes some abnormalities in spermatozoa but freezing without cryoprotection does not and 2) long-term storage of both frozen and freeze-dried spermatozoa is not deleterious to their genetic integrity. Freezing without cryoprotection is highly successful, simple, and efficient but, like all routine sperm storage methods, requires liquid nitrogen. Liquid nitrogen is also required for freeze-drying, but sperm can then be stored at 4 degrees C and shipped at ambient temperatures. Both preservation methods are successful, but rapid freezing without cryoprotection is the preferred method for preservation of spermatozoa from mouse strains carrying unique genes and mutations.

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“…It is also not caused by the inability of these sperm to undergo the acrosome reaction (AR), because sperm from t/t males undergo both the spontaneous AR and the zona-induced AR at rates and levels similar to sperm from congenic wildtype males (Olds-Clarke and Johnson 1993;Johnson et al 1995a). Sperm from t/t males are also able to form normal pronuclei and participate in the formation of normal blastocysts after intracytoplasmic injection into mouse eggs (Kuretake et al 1996). Furthermore, since at least some of the sperm from t/t males can bind to the oocyte membrane (Johnson et al 1995a), the major defect appears to be in the fusion phase of sperm-oolemma penetration.…”

Section: Introductionmentioning

confidence: 99%

High-resolution mapping of sperm function defects in the t complex fourth inversion

et al. 1998

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Structural variants of the mouse Chr 17-specific t complex, known as t haplotypes, express factors that alter the ability of sperm to carry out their roles in the normal fertilization process. In previous studies of males carrying heterospecific combinations of the t complex, we discovered a unique M. spretus/t haplotype phenotype of male sterility. In additional studies with mice carrying a series of M. spretus-M. m. domesticus recombinant Chr 17 homologs and a complete t haplotype (S-+/t), we monitored physiological aspects of sperm function to map a locus (Hst6) responsible for expression of the t-specific "curlicue" sperm flagellar curvature phenotype to 1 cM within the fourth inversion of the t complex. In the present report, we quantitatively analyze the in vitro capability of sperm from mice with similar S-+/t Chr 17 genotypes to fertilize zona pellucida-free mouse eggs. The results identify a locus, Stop1, mapping distal to Pim1, with acute effects on the ability of sperm to penetrate the oolemma. The data suggest that Stop1 is a complex locus consisting of at least two genetic elements, a proximal one overlapping the Hst6 locus, and another, distal to the Hst6 locus. Further quantitative analyses of the "curlicue" phenotype produced by sperm derived from these same animals indicate that expression of this chronic flagellar curvature phenotype also derives from at least two elements, both mapping within the Hst6 locus. Thus, these studies provide higher resolution mapping of the molecular basis of t haplotype-specific sperm dysfunction emanating from In(17)4.

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Inadequate function of sterile tw5/tw32 spermatozoa overcome by intracytoplasmic sperm injection

Cited by 13 publications

References 0 publications

Intracytoplasmic Sperm Injection Effects in Infertile azh Mutant Mice1

Intracytoplasmic Sperm Injection Effects in Infertile azh Mutant Mice1

Long-Term Preservation of Mouse Spermatozoa after Freeze-Drying and Freezing Without Cryoprotection1

High-resolution mapping of sperm function defects in the t complex fourth inversion

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