Sperm Sexing in the Dog by Fluorescence In Situ Hybridization.
Notice bibliographique
Résumé
The specific production of male or female offspring by separation of X- or Y-bearing spermatozoa and artificial insemination is a long standing goal in breeding of domestic and other animals. For dogs, higher percentages of female puppies are desired in breeding of guide dogs for the blind as well as companion animals. Effective preselection of sex has been accomplished in several livestock and humans using flow-cytometric sperm sorting. Since a guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing, to be able to apply the specific production of male or female in dog, direct visualization of sex chromosomes in single sperm using fluorescence in situ hybridization (FISH) was developed in this study. The second fraction of ejaculates was collected from Labrador Retrievers (n=3). After washing with PBS, the sperms were fixed with methanol: acetic acid fixative and placed on a defatted glass slide. These preparations were dried and dehydrated on a hot plate at 70°C. For decondensation of sperm nuclei, the specimen was treated with 1M NaOH for 3, 4 or 5 min at room temperature, rinsed with PBS, and then washed with distilled water. After air-drying, the specimen was placed on a hot plate at 70°C. For a double staining of canine X- and Y-chromosome of sperm, sex chromosome probe: X-labelled with SpecrumGreen and Y-labelled with Cy3 were used. Hybridization was carried out in a moist chamber at 37°C overnight. FISH signals were analyzed by eye using an ocular grid counting random microscope fields and scoring 500-600 spermatozoa per sample. Spermatozoa were scored only if they were intact, non-overlapped, had a clearly defined border, and had not decondensed to more than twice the size of a non-decondensed sperm head. There were no significant difference at the P>0.05 level between the theoretical ratio (50:50) and the observed ratio in X- and Y-chromosome-bearing spermatozoa from all three dogs, when samples were treated with 1 M NaOH for 4 min. Hybridization efficiency was 98-99%. The established FISH protocol in this study might be suitable for reliable routine validation of sorted canine spermatozoa.
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».