Clonal analysis of cardiac morphogenesis in the chicken embryo using a replication‐defective retrovirus: I. Formation of the ventricular myocardium
Clonal analysis of cardiac morphogenesis in the chicken embryo using a replication‐defective retrovirus: I. Formation of the ventricular myocardium
复制标题
使用复制缺陷型逆转录病毒对鸡胚心脏形态发生进行克隆分析：I. 心室心肌的形成
作者：
T. Mikawa;A. Borisov;A. Brown;D. Fischman
心前中胚层细胞（4-6期）和早期心脏（10-15期）的分裂肌细胞用一种编码细菌β -半乳糖苷酶（β - gal）的复制无能逆转录病毒（CXL） （Mikawa et al., 1991 b）进行标记。用两种方法感染致心细胞。(1)从原胚期胚胎（4-6期）上切下小块（~ 50 μm2）的前外侧中胚层，在含逆转录病毒的液体培养基中培养。去除CXL后，将组织分散成单细胞悬浮液，并将压力注射到受体胚胎的心前区（4-6期）。然后将这些胚胎在体外37°C孵育2天（New, 1968），将具有跳动心脏的胚胎固定，进行X - gal组织化学和石蜡连续切片。(2)将卵（胚胎期4-15期）内的CXL加压注射到致心组织中，随后将卵送回培养箱。在选定的发育阶段，固定胚胎或整个心脏，用X - gal染色，石蜡包埋后连续切片。第一种方法表明：(1)逆转录病毒可感染心前中胚层细胞，(2)移植细胞可分化为跳动的肌细胞，(3)β - gal表达量高，可进行组织化学检测。通过第二种方法，我们可以发现：(1)β - gal标记的细胞在心肌中形成集落，(2)标记的细胞完全是肌细胞，(3)每个集落的细胞数量随着胚胎发育年龄的增加而增加，(4)左心室集落的大小比右心室大，(5)许多集落是跨壁的，即，它们从心外膜延伸到心内膜，通常呈锥状或漏斗状，锥的基部最靠近心外膜，(6)每个集落内肌细胞的方向在心肌的不同层发生变化，(7)锥体同时含有β - gal+和β - gal -肌细胞。用[3H]胸苷进行的DNA标记研究表明，在发育的第一周，心源性细胞每16-18小时分裂一次，CXL标记的细胞与未标记的肌细胞分裂无明显区别。在此基础上，提出了心肌生长的模型。
Cells of the precardiac mesoderm (stages 4–6) and dividing myocytes of early hearts (stages 10–15) were tagged with a replication‐incompetent retrovirus (CXL) (Mikawa et al., 1991 b) encoding bacterial β‐galactosidase (β‐gal). Two protocols were used to infect the cardiogenic cells. (1) Small blocks (∼50 μm2) of anterolateral mesoderm were dissected from gastrula‐stage embryos (stages 4–6) and incubated in liquid medium containing the retrovirus. After removal of CXL, the tissues were dispersed into single‐cell suspensions and pressure injected into the precardiac areas of recipient embryos (stages 4–6). Such embryos were then incubated in vitro at 37°C for 2 days (New, 1968), and those embryos with beating hearts were fixed for X‐gal histochemistry and paraffin serial sectioning. (2) CXL was pressure injected in ovo (embryonic stages 4–15) into cardiogenic tissues and the eggs subsequently returned to an incubator. At selected stages of development embryos or whole hearts were fixed, stained with X‐gal, and serially sectioned after paraffin embedding. The first method showed that (1) cells of the precardiac mesoderm could be infected with the retrovirus, (2) the transplanted cells would differentiate into beating myocytes, and (3) β‐gal expression was sufficiently high to be detected histochemically. With the second procedure we could show that (1) β‐gal‐tagged cells formed colonies in the myocardium, (2) the labeled cells were exclusively myocytes, (3) the number of cells per colony increased with increasing age of embryonic development, (4) the size of colonies was larger in the left than the right ventricle, (5) many of the colonies were transmural, i.e., they extended from epicardial to endocardial layers of the myocardium and generally exhibited a cone or funnel‐shape with the base of the cone nearest the epicardium, (6) the orientation of myocytes within each colony changed at different layers of the myocardium, and (7) the cones contained both β‐gal+ and β‐gal− myocytes. DNA labeling studies with [3H]thymidine indicated that cardiogenic cells divided every 16–18 hr during the first week of development and that the CXL‐labeled cells divided indistinguishably from unlabeled myocytes. Based on these observations a model for the growth of the myocardium is presented.
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横纹肌结缔组织的功能排列，重点是心肌。
DOI：
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发表时间：
1982
期刊：
Scanning electron microscopy
影响因子：
--
作者：
Borg,TK;Sullivan,T;Ivy,J
通讯作者：
Ivy,J
胚胎鸡心脏中包膜病毒的极化释放：假定心肌中上皮极性的证明。
影响因子：
2.7
作者：
Peng,I;Dennis,JE;Rodriguez-Boulan,E;Fischman,DA
通讯作者：
Fischman,DA
影响因子：
56.9
作者：
WALSH, C;CEPKO, CL
通讯作者：
CEPKO, CL
心肌细胞与细胞外基质成分的粘附。
影响因子：
9.5
作者：
Lundgren,E;Terracio,L;Borg,TK
通讯作者：
Borg,TK