Durcan Peter Joseph, Conradie Johannes D, Van deVyver Mari, Myburgh Kathryn Helen
Department of Physiological Science, Stellenbosch University, Private Bag X1 Matieland, 7602, Stellenbosch, South Africa.
Division of Endocrinology, Department of Medicine, Stellenbosch University, Tygerberg, South Africa.
BMC Physiol. 2014 Dec 9;14:11. doi: 10.1186/s12899-014-0011-3.
Multiple cell types including trophoblasts, osteoclasts and myoblasts require somatic cell fusion events as part of their physiological functions. In Drosophila Melanogaster the paralogus type 1 transmembrane receptors and members of the immunoglobulin superfamily Kin of Irre (Kirre) and roughest (Rst) regulate myoblast fusion during embryonic development. Present within the human genome are three homologs to Kirre termed Kin of Irre like (Kirrel) 1, 2 and 3. Currently it is unknown if Kirrel3 is expressed in adult human skeletal muscle.
We investigated (using PCR and Western blot) Kirrel3 in adult human skeletal muscle samples taken at rest and after mild exercise induced muscle damage. Kirrel3 mRNA expression was verified by sequencing and protein presence via blotting with 2 different anti-Kirrel3 protein antibodies. Evidence for three alternatively spliced Kirrel3 mRNA transcripts in adult human skeletal muscle was obtained. Kirrel3 mRNA in adult human skeletal muscle was detected at low or moderate levels, or not at all. This sporadic expression suggests that Kirrel3 is expressed in a pulsatile manner. Several anti Kirrel3 immunoreactive proteins were detected in all adult human skeletal muscle samples analysed and results suggest the presence of different isoforms or posttranslational modification, or both.
The results presented here demonstrate for the first time that there are at least 3 splice variants of Kirrel3 expressed in adult human skeletal muscle, two of which have never previously been identified in human muscle. Importantly, mRNA of all splice variants was not always present, a finding with potential physiological relevance. These initial discoveries highlight the need for more molecular and functional studies to understand the role of Kirrel3 in human skeletal muscle.
包括滋养层细胞、破骨细胞和成肌细胞在内的多种细胞类型需要体细胞融合事件作为其生理功能的一部分。在黑腹果蝇中,同源的1型跨膜受体以及免疫球蛋白超家族的Irre相关激酶(Kirre)和最粗糙蛋白(Rst)成员在胚胎发育过程中调节成肌细胞融合。人类基因组中存在三个与Kirre同源的基因,即Irre样激酶(Kirrel)1、2和3。目前尚不清楚Kirrel3是否在成年人类骨骼肌中表达。
我们(使用PCR和蛋白质印迹法)研究了成年人类骨骼肌样本在静息状态以及轻度运动诱导肌肉损伤后的Kirrel3。通过测序验证了Kirrel3 mRNA的表达,并使用两种不同的抗Kirrel3蛋白抗体通过印迹法检测到了蛋白质的存在。获得了成年人类骨骼肌中三种选择性剪接的Kirrel3 mRNA转录本的证据。成年人类骨骼肌中的Kirrel3 mRNA检测到低水平或中等水平表达,或者根本未检测到。这种散在的表达表明Kirrel3以脉冲方式表达。在所有分析的成年人类骨骼肌样本中检测到了几种抗Kirrel3免疫反应性蛋白,结果表明存在不同的异构体或翻译后修饰,或两者皆有。
此处呈现的结果首次证明,成年人类骨骼肌中表达至少3种Kirrel3剪接变体,其中两种此前从未在人类肌肉中被鉴定出来。重要的是,并非所有剪接变体的mRNA都始终存在,这一发现具有潜在的生理相关性。这些初步发现凸显了需要更多分子和功能研究来了解Kirrel3在人类骨骼肌中的作用。