Use of somatic cells from goat milk for dynamic studies of gene expression in the mammary gland.

摘要: Somatic cells are present in the milk throughout lactation and consist of leukocytes epithelial exfoliated from mammary epithelium. Our objective was to determine efficacy using somatic goat for dynamic studies gene expression gland. Over a 4-wk interval, were isolated daily morning samples taken 30 min after milking. They characterized by direct cell counts flow cytometry analysis immunostaining with antibodies directed against cytokeratin CD45, common leukocyte antigen. Epithelial within ranged 15 45% total cells. After-milking contained twice as many did samples. The RNA extracted both types equivalent efficiency (a mean 1.2 microg RNA/mL milk). Four mRNA variants alpha-S1 casein detected Northern blot amount each related protein concentration milk. comparison between gland congruently collected showed that relative amounts milk-protein (alpha-S1 kappa-casein alactalbumin) conserved. In third experiment, preparations assess effect growth hormone (GH) on expression; four goats separated into two groups order perform switch-back design consisting three treatment weeks: Control, GH-Control or GH-Control-GH. this study, GH increased yields 5%. Throughout control treatments, genes studied highly significantly correlated (r = 0.949 r 0.958, P < 0.001 for, respectively, alpha-lactalbumin). During treatment, abundances same pattern. conclusion, opportunity use preparation provides significant improvement over biopsy assessing activity allows easy repetitive sampling without damaging tissue. Furthermore, we propose method could be used investigate transcriptional status an animal relation its genotype, nutritional pathologic status, under influence hormonal factors.