Department of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg 24061, USA.
J Dairy Sci. 2011 Oct;94(10):5005-16. doi: 10.3168/jds.2011-4229.
Pubertal mammary gland growth and development are hormonally regulated, but the details are poorly understood in calves. Our purpose was to evaluate the effects of exogenous growth hormone (GH) on the biochemical composition of the prepubertal mammary gland, mRNA expression of selected genes, and histological characteristics of the developing parenchyma (PAR). In this experiment, 19 calves (7 ± 4 d of age) were randomly assigned to 1 of 2 treatments: bovine somatotropin (bST, 500 mg; n = 10) or placebo (Sal; 0.9% saline; n = 9). Animals were treated every 3 wk beginning on d 23. Calves were assigned to an early (65 d; tissue harvested after 2 treatment injections) or late collection time (107 d; tissue harvested after 4 treatment injections). Calves were fed milk replacer and calf starter for 8 wk and starter and hay thereafter. Parenchyma and mammary fat pad (MFP) from one udder half were harvested for analysis of protein, lipid, and DNA. Additional tissues were preserved for histological analysis or snap-frozen for quantitative real-time PCR. Somatotropin treatment did not significantly alter the mass of PAR or MFP or the general pattern of development of epithelial structures. Significant increases were observed in protein/100 kg of body weight (BW), total protein, DNA concentration, DNA/100 kg of BW, and total DNA in 107-d calves, and a significant treatment by day interaction was observed for DNA and lipid concentrations in PAR. In MFP, a significant decrease was observed in protein/100 kg of BW in bST-treated calves and in total MFP protein in 65-d calves. A treatment by day interaction was found for total protein, DNA, and protein/100 kg of BW. In PAR, relative expression of ATPase-binding cassette 3 and growth hormone receptor were reduced by bST and both were lower in 107-d-harvest calves. Epithelial cell retention of bromodeoxyuridine (BrdU; possible indicator of stem-like cells) was greatest in 65-d bST-treated calves, and a significant time of sampling response and treatment × time interaction were observed. Expression of the proliferation marker protein Ki67 was numerically higher in bST-treated calves but the difference was nonsignificant. Retention of the BrdU label was reduced in 107-d calves. Exogenous growth hormone given to calves may affect mammary tissue composition and epithelial cell gene expression in subtle ways but exogenous supplementation with bST alone is not likely to alter overall development patterns or affect the mass of mammary parenchymal tissue. Whether such subtle changes have an effect on subsequent development or function is unknown.
青春期乳腺的生长和发育受激素调控,但小牛的具体细节尚不清楚。我们的目的是评估外源性生长激素(GH)对未成熟乳腺的生化组成、选定基因的 mRNA 表达以及发育实质(PAR)组织学特征的影响。在这项实验中,19 头小牛(23 日龄时为 7±4 日龄)被随机分配到以下 2 种处理中的 1 种:牛生长激素(bST,500mg;n=10)或安慰剂(Sal;0.9%生理盐水;n=9)。动物从第 23 天开始每 3 周接受 1 次治疗。小牛被分配到早期(65 天;接受 2 次治疗注射后采集组织)或晚期采集时间(107 天;接受 4 次治疗注射后采集组织)。小牛在 8 周龄时饲喂代乳料和小牛饲料,之后饲喂饲料和干草。一半的乳腺组织和乳腺脂肪垫(MFP)用于分析蛋白质、脂质和 DNA。其余组织用于组织学分析或用于实时定量 PCR 的冻存。生长激素处理并未显著改变 PAR 或 MFP 的质量或上皮结构发育的一般模式。107 日龄小牛的蛋白质/100kg 体重(BW)、总蛋白、DNA 浓度、DNA/100kg BW 和总 DNA 显著增加,PAR 中 DNA 和脂质浓度观察到处理与日龄的显著交互作用。在 MFP 中,bST 处理的小牛的蛋白质/100kg BW 中蛋白质和 65 日龄小牛的总 MFP 蛋白显著减少。总蛋白、DNA 和蛋白质/100kg BW 中观察到处理与日龄的交互作用。在 PAR 中,ATP 结合盒转运蛋白 3 和生长激素受体的相对表达被 bST 降低,并且在 107 日龄收获的小牛中均较低。溴脱氧尿苷(BrdU;可能的干细胞标志物)在 65 日龄 bST 处理的小牛中的保留最多,并且观察到采样时间和处理与时间的显著交互作用。Ki67 增殖标志物蛋白的表达在 bST 处理的小牛中数值较高,但差异无统计学意义。107 日龄小牛的 BrdU 标记保留减少。给小牛注射外源性生长激素可能会以微妙的方式影响乳腺组织成分和上皮细胞基因表达,但单独补充外源性 bST 不太可能改变整体发育模式或影响乳腺实质组织的质量。这种微妙的变化是否会对随后的发育或功能产生影响尚不清楚。