Preliminary results of hemizona assay (HZA) as a fertility test for canine spermatozoa
TL;DR: The objective of this study was to validate the HZA for canine spermatozoa, as a test for diagnosing canine male fertility status, and to avoid the effect of bad zona on sperm binding.
About: This article is published in Theriogenology.The article was published on 1998-07-15. It has received 30 citations till now.
Citations
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TL;DR: Improvement of IVM, in vitro fertilization (IVF) and embryo culture coupled with ovarian tissue grafting, cryobanking of follicles, oocytes, semen, or embryos, with subsequent ET into surrogate females, may render this technology feasible for use in endangered wild felids.
167 citations
TL;DR: Assays based on in vitro fertilization (IVF) and related techniques such as zona pellucida (ZP) binding assays and oocyte penetration tests, which can provide valuable information about a semen donor, an insemination dose or a method of semen preservation are presented.
157 citations
TL;DR: Although substantial improvements have been made in canine semen assessment, surprisingly few parameters were correlated with in vivo fertility, therefore, further research is required to determine which sperm characteristics are of clinical value for predicting the in vivoertility in dogs.
92 citations
Cites background or methods from "Preliminary results of hemizona ass..."
...The zona binding capacity of canine spermatozoa has been evaluated by two types of assays: one using intact homologous oocytes (ZP-binding assay, ZBA) [66,68,69], the other using bisected hemizonae (hemizona binding assay, HZA) [13,67]....
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...The two parts of the zona are subsequently incubated with a semen sample and the number of bound spermatozoa is determined using phasecontrast microscopy [13,67]....
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...Using this assay, the canine oocyte is bisected by means of micromanipulation and the dense ooplasma is dislodged [13,67]....
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...The advantages of the HZA are the possibility to compare the binding capacities between a control male and a test male, and that the zonae may be stored either chilled or frozen [13]....
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...However, the HZA is time-consuming and technically more demanding [68], and various factors may affect the results, such as the quality of the zona and the concentration of motile spermatozoa used in the assay [13]....
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TL;DR: Some of the classical and new methods currently used for dog sperm evaluation are reviewed, though not exhaustively, covering some aspects of the sperm cell that are useful in estimating its functional ability.
85 citations
TL;DR: Results show that a commercial system for extending, chilling and transporting equine semen is an attractive and efficient method of shipping canine extended chilled semen.
51 citations
References
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TL;DR: Sperm from fertile men exhibited significantly higher binding capacity to hemizonae compared with sperm from men who had fertilization failure during in vitro fertilization (IVF) treatment, demonstrating that the HZA may be a useful diagnostic tool in male infertility evaluations.
390 citations
TL;DR: It is demonstrated that intact and solubilized canine ZP are capable of inducing acrosome reactions of canine sperm.
Abstract: In this study the induction of the acrosome reaction of canine sperm by homologous zona pellucida (ZP) was examined. Twelve semen samples obtained from 6 normal beagle dogs were evaluated after sperm incubation in vitro with canine capacitation medium (CCM). Washed sperm were preincubated at 37 degrees C in 5% CO2 in air for 4 and 7 h prior to experimental treatment. Sperm were co-incubated for 1 min with intact oocytes collected from canine ovaries. Half of the oocytes were then fixed, and the bound sperm were assessed for acrosome reactions through use of a polyclonal antisperm antiserum and indirect immunofluorescence. The remaining oocytes were incubated in sperm-free medium for an additional 1-h period, and the acrosomal status of sperm bound to the ZP was evaluated similarly. The percentage of acrosome-reacted sperm on the ZP increased significantly during the 1-h incubation period. In other experiments, capacitated canine sperm were incubated with heat-solubilized ZP for 1 h and their acrosomal status was determined using fluoresceinated Pisum sativum lectin. The percentages of acrosome-reacted sperm increased significantly in ZP solution compared with controls. These data demonstrate that intact and solubilized canine ZP are capable of inducing acrosome reactions of canine sperm.
77 citations
TL;DR: The findings show that the sequential application of bioassays can elucidate specific sperm or oocyte defects that characterize functional abnormalities among sperm populations.
76 citations
TL;DR: It was concluded that the HZA is a good predictor of fertilization rate in vitro, and can be used in the IVF setting to supply additional clinical information in male-factor patients.
Abstract: The objective of this prospective study was to examine the ability of the hemizona assay (HZA) to predict fertilization outcome of mature, pre-ovulatory oocytes under in-vitro fertilization (IVF) conditions. Since a large number of patients were evaluated over a long period, the power of the HZA to prognosticate fertilization results in the same and subsequent (consecutive) IVF cycles of those same patients was assessed. For IVF, only metaphase II oocytes were used. For the HZA, both fresh oocytes donated by patients at the time of IVF and oocytes recovered from surgically removed ovarian tissue (and salt-stored) were used, and bisected by micromanipulation techniques. Matching hemizonae were co-incubated either with spermatozoa from the patient (test) or from a fertile man (control) for 4 h. The number of spermatozoa tightly bound to the zona was counted. Patients (n = 112) were divided into two groups based on HZA results (expressed as HZA index or HZI): HZI > or = 30% (n = 72) and or = 30% (P < 0.03). Linear discriminant analysis indicated the HZA to have a sensitivity of 84%, and positive and negative predictive values of 85 and 70% respectively, for prediction of fertilization outcome in a total of 233 cycles. It was concluded that the HZA is a good predictor of fertilization rate in vitro, and can be used in the IVF setting to supply additional clinical information in male-factor patients.
63 citations
TL;DR: The influence of sperm morphologic pattern as a prognosticator of fertilization outcome is established in major part as a determinant of sperm binding ability to the zona pellucida and the hemizona assay provides a robust index that is highly predictive of the potential of human gametes to achieve fertilization and highlights its use in infertility and contraception testing.
59 citations