Comparative analysis of hypotonic solutions for cytogenetic studies of animals

Abstract

Relevance. Cytogenetic studies are in demand to identify carriers of hereditary anomalies. In order to obtain metaphase plates of the chromosomes of various animal species, we optimized the usual method, taking into account some adjustments. Methods. The main research criterion was the selection of a hypotonic solution for cytogenetic studies in sheep and goats. 3 variants of using hypotonic solutions were considered: 0.56% solution of potassium chloride; distilled water and 0.56% solution of potassium chloride; 2% solution of sodium citrate and 0.56% solution of potassium chloride (1:1). For the cultivation of peripheral blood cells, blood was taken from the jugular vein of animals (goats n = 8, sheep n = 11). Results. The cells were cultured according to the generally accepted method. The results of the conducted studies showed that the quality of the preparations differed depending on the components used. The optimal parameters for the preparation of chromosomes with a good distribution were obtained by culturing sheep lymphocyte cells using 0.56% potassium chloride as a hypotonic solution (20 min) and ranged from 14.29 to 25.00%. For cytogenetic studies in goats, optimal results were found when using a 2% solution of sodium citrate with a 0.56% solution of potassium chloride (1:1) for 20 minutes — from 13.33 to 25.00%. Each option allowed to reduce the loss of cells during the preparation of chromosome preparations for a certain type of animal. Thus, the developed protocol for obtaining metaphase plates using a 2% solution of sodium citrate with a 0.56% solution of potassium chloride (1:1) as a hypotonic solution is optimal when used in goats, while the classical method (0.56% solution of potassium chloride) was most suitable for sheep.