Killian Christopher E, Croker Lindsay, Wilt Fred H
Department of Molecular and Cell Biology, University of California, Berkeley, 142 Life Sciences Addition, MC# 3200, Berkeley, CA 94720-3200, USA.
Gene Expr Patterns. 2010 Feb-Mar;10(2-3):135-9. doi: 10.1016/j.gep.2010.01.002. Epub 2010 Jan 25.
The SpSM30 gene family of the sea urchin, Strongylocentrotus purpuratus, is comprised of six members, designated SpSM30A through SpSM30F (Livingston et al., 2006). The SpSM30 proteins are found uniquely in embryonic and adult mineralized tissues of the sea urchin. Previous studies have revealed that SpSM30 proteins are occluded within the embryonic endoskeleton and adult mineralized tissues (Killian and Wilt, 1996; Mann et al., 2008a,b; Urry et al., 2000). Furthermore, some of the SpSM30 proteins are among the most abundant of the approximately four-dozen integral matrix proteins of the larval spicule (Killian and Wilt, 1996). The amino acid sequence, protein domain architecture, and contiguity within the genome strongly support the supposition that the six genes constitute a gene family. Reverse transcription-polymerase chain reaction (RT-PCR) is used in the present study to describe the time course of expression of the family members during embryonic development, and their expression in adult tissues. SpSM30A, B, C and E are expressed, albeit at different levels, during overt spicule deposition in the embryo with some differences in the precise timing of expression. SpSM30D is not expressed in the embryo, and SpSM30F is expressed transiently and at low levels just prior to overt spicule formation. Whole mount in situ hybridization studies show that SpSM30A, B, C, and E are expressed exclusively in primary mesenchyme (PMC) cells and their descendants. In addition, tissue fractionation studies indicate that SpSM30F expression is highly enriched in PMCs. Each adult tissue examined expresses a different cohort of the SpSM30 family members at varying levels: SpSM30A mRNA is not expressed in adult tissues. Its expression is limited to the embryo. Conversely, SpSM30D mRNA is not expressed in the embryo, but is expressed in adult spines and teeth. SpSM30B and SpSM30C are expressed at modest levels in all mineralized adult tissues; SpSM30E is expressed highly in tooth and test; and SpSM30F is expressed in spine and at low levels in the other adult tissues except the test. Relative levels of expression of the several family members in these different tissues vary widely. It is likely SpSM30 proteins play a vital, but still unknown, role in biomineralization of these tissues during development.
紫球海胆(Strongylocentrotus purpuratus)的SpSM30基因家族由六个成员组成,命名为SpSM30A至SpSM30F(利文斯顿等人,2006年)。SpSM30蛋白仅在海胆的胚胎和成年矿化组织中发现。先前的研究表明,SpSM30蛋白被封闭在胚胎内骨骼和成年矿化组织中(基利安和威尔特,1996年;曼等人,2008年a、b;厄里等人,2000年)。此外,一些SpSM30蛋白是幼虫骨针约四十多种整合基质蛋白中含量最丰富的蛋白之一(基利安和威尔特,1996年)。氨基酸序列、蛋白质结构域结构以及基因组内的相邻关系有力地支持了这六个基因构成一个基因家族的假设。本研究采用逆转录聚合酶链反应(RT-PCR)来描述家族成员在胚胎发育过程中的表达时间进程及其在成年组织中的表达情况。SpSM30A、B、C和E在胚胎明显的骨针沉积过程中均有表达,尽管表达水平不同,在表达的精确时间上也存在一些差异。SpSM30D在胚胎中不表达,而SpSM30F在明显的骨针形成之前短暂且低水平表达。整体原位杂交研究表明,SpSM30A、B、C和E仅在初级间充质(PMC)细胞及其后代中表达。此外,组织分级分离研究表明,SpSM30F的表达在PMC中高度富集。所检测的每个成年组织均以不同水平表达SpSM30家族成员的不同组合:SpSM30A mRNA在成年组织中不表达,其表达仅限于胚胎。相反,SpSM30D mRNA在胚胎中不表达,但在成年棘和牙齿中表达。SpSM30B和SpSM30C在所有成年矿化组织中适度表达;SpSM30E在牙齿和围口膜中高表达;SpSM30F在棘中表达,在除围口膜外的其他成年组织中低表达。这几个家族成员在这些不同组织中的相对表达水平差异很大。SpSM30蛋白在这些组织发育过程中的生物矿化中可能起着至关重要但仍不清楚的作用。