Koff M D, Plaut K
Department of Animal and Food Sciences, University of Vermont, Burlington 05405, USA.
J Dairy Sci. 1995 Sep;78(9):1903-8. doi: 10.3168/jds.S0022-0302(95)76815-1.
The objective of this study was to evaluate the expression of mRNA for transforming growth factor-alpha in the bovine mammary gland. Total RNA from 23 cows at various stages of development was isolated from mammary parenchyma and subjected to Northern blot analysis using human transforming growth factor-alpha 32P-labeled deoxycytosine triphosphate cDNA probe. When blots were washed through 1x sodium chloride and sodium citrate plus 1% SDS at 60 degrees C, a 4.8-kb band was observed in all RNA samples. When stringency was increased to .1x, mRNA transcripts were observed only for cows at d 233, 240, and 244 of gestation. After the high stringency washes, major bands were observed at 7.4 and 3.3 kb and minor bands on d 233 at 5.4, 2.3, and 1.6 kb. No additional transcripts were detected in RNA from mammary glands of nonpregnant cows during involution, lactating cows, or estrogen-treated heifers. These data indicate that transcripts similar to transforming growth factor-alpha are present in the bovine mammary gland. The expression pattern could represent alternatively spliced transcripts that were produced during an important period of mammary development prior to parturition.
本研究的目的是评估转化生长因子-α的mRNA在牛乳腺中的表达。从处于不同发育阶段的23头奶牛的乳腺实质中分离出总RNA,并使用人转化生长因子-α 32P标记的脱氧胞苷三磷酸cDNA探针进行Northern印迹分析。当印迹在60℃下用1×氯化钠和柠檬酸钠加1%十二烷基硫酸钠洗涤时,在所有RNA样品中均观察到一条4.8kb的条带。当严谨度提高到0.1×时,仅在妊娠第233、240和244天的奶牛中观察到mRNA转录本。经过高严谨度洗涤后,在第233天观察到7.4和3.3kb的主要条带以及5.4、2.3和1.6kb的次要条带。在非妊娠奶牛乳腺退化期、泌乳期奶牛或雌激素处理的小母牛的RNA中未检测到其他转录本。这些数据表明,牛乳腺中存在与转化生长因子-α相似的转录本。这种表达模式可能代表在分娩前乳腺发育的重要时期产生的可变剪接转录本。