Buff P R, Dodds A C, Morrison C D, Whitley N C, McFadin E L, Daniel J A, Djiane J, Keisler D H
Animal Sciences Department, University of Missouri, Columbia 65211, USA.
J Anim Sci. 2002 Nov;80(11):2942-8. doi: 10.2527/2002.80112942x.
Obesity has been a major concern in the horse industry for many years, and the recent discovery of leptin and leptin receptors in numerous nonequine species has provided a basis for new approaches to study this problem in equine. The objectives were to: 1) clone a partial sequence ofthe equine leptin and leptin receptor genes so as to enable the design of primers for RT-PCR determination of leptin and leptin receptor gene presence and distribution in tissues, 2) develop a radioimmunoassay to quantify peripheral concentrations of leptin in equine, 3) determine if peripheral concentrations of leptin correlate with body condition scores in equine, and 4) determine if changing body condition scores would influence peripheral concentrations of leptin in equine. In Experiment 1, equine leptin (GenBank accession number AF179275) and the long-form of the equine leptin receptor (GenBank accession number AF139663) genes were partially sequenced. Equine leptin receptor mRNA was detected in liver, lung, testis, ovary, choroid plexus, hypothalamus, and subcutaneous adipose tissues using RT-PCR. In Experiment 2, 71 horses were categorized by gender, age, and body condition score and blood samples were collected. Sera were assayed for leptin using a heterologous leptin radioimmunoassay developed for equine sera. Serum concentrations of leptin increased in horses with body condition score (1 = thin to 9 = fat; r = 0.64; P = 0.0001). Furthermore, serum concentrations of leptin were greater in geldings and stallions than in mares (P = 0.0002), and tended to increase with age of the animal (P = 0.08). In Experiment 3, blood samples, body weights, and body condition scores were collected every 14 d from 18 pony mares assigned to gain or lose weight over a 14-wk interval based on initial body condition score. Although statistical changes (P = 0.001) in body condition scores were achieved, congruent statistical changes in peripheral concentrations of leptin were not observed, likely due to the small range of change that occurred. Nonetheless, serum concentrations of leptin tended to be greater in fat-restricted mares than in thin-supplemented mares (P = 0.09). We conclude that leptin and leptin receptors are present in equine tissues and that peripheral concentrations of leptin reflect a significant influence of fat mass in equine.
多年来,肥胖一直是马匹行业的一个主要问题,最近在许多非马属物种中发现了瘦素和瘦素受体,为在马属动物中研究这个问题的新方法提供了基础。目标是:1)克隆马属动物瘦素和瘦素受体基因的部分序列,以便设计引物用于RT-PCR测定瘦素和瘦素受体基因在组织中的存在和分布;2)开发一种放射免疫分析法来定量马属动物外周血中瘦素的浓度;3)确定马属动物外周血中瘦素的浓度是否与身体状况评分相关;4)确定改变身体状况评分是否会影响马属动物外周血中瘦素的浓度。在实验1中,对马属动物瘦素(GenBank登录号AF179275)和马属动物瘦素受体的长形式(GenBank登录号AF139663)基因进行了部分测序。使用RT-PCR在肝脏、肺、睾丸、卵巢、脉络丛、下丘脑和皮下脂肪组织中检测到马属动物瘦素受体mRNA。在实验2中,根据性别、年龄和身体状况评分对71匹马进行分类,并采集血样。使用为马属动物血清开发的异源瘦素放射免疫分析法检测血清中的瘦素。身体状况评分(1=瘦至9=胖)的马血清中瘦素浓度升高(r=0.64;P=0.0001)。此外,去势公马和种马血清中的瘦素浓度高于母马(P=0.0002),并且倾向于随着动物年龄的增长而增加(P=0.08)。在实验3中,根据初始身体状况评分,从18匹母马中每14天采集一次血样、体重和身体状况评分,这些母马被分配在14周的时间间隔内增重或减重。尽管身体状况评分有统计学变化(P=0.001),但未观察到外周血中瘦素浓度有相应的统计学变化,可能是由于变化范围较小。尽管如此,限食母马血清中的瘦素浓度往往高于瘦马补充营养后的母马(P=0.09)。我们得出结论,瘦素和瘦素受体存在于马属动物组织中,并且外周血中瘦素的浓度反映了脂肪量对马属动物的显著影响。