Analysis of calcium channels involved in craniofacial neural crest cell migration

为了检测电压门控钙通道α1亚基A、B、C、D和E亚型抗体的特异性，我们对出生后0、7、14天和成年小鼠脑石蜡切片进行了免疫组织化学染色。α1亚基的A、B、C、D、E亚型在浦肯野细胞上均有表达。我们使用原位杂交技术的前一个结果支持了这个结果。抗体对α1亚基的A、B、C、D、E亚型具有良好的特异性。结果表明，电压门控钙通道α1亚基的表型可能不依赖于细胞种类。鸡胚神经管标记神经上皮或抗HNK-1抗体免疫组化染色显示神经嵴细胞在中胚层区迁移。固定前应用鸡胚BrdU免疫组化染色检测神经嵴细胞S期。1)迁移神经嵴的中胚层细胞在迁移神经嵴的途中增殖。免疫组化染色检测小鼠胚胎第8 ~ 9.5天电压门控钙通道α1亚基的A、B、C、D、E亚型。2) 5种类型中α1亚基C、D型在神经嵴细胞迁移开始时强烈表达。3)迁移过程中表达了C、D、E型。4)胚胎第8天，神经管神经上皮中未见α1亚基的表达，胚胎第9.5天，α1亚基的表达呈分散状态。这些结果表明，电压门控钙通道参与了神经嵴细胞从迁移开始到迁移过程中的迁移。4)神经上皮上的电压门控钙通道既不分泌也不传递神经，有新的功能。

In order to examine the specification of antibodies to A, B, C, D and E subtypes of voltage-gated calcium channel α1 subunit, we did immunohistochemical staining on paraffin sections of the mouse brain at postnatal day 0, 7, 14 and adult. A, B, C, D and E subtypes of α1 subunit were expressed on Purkinje cells. The former result of us using in situ hybridization technique supported the result. The antibodies have good specification to A, B, C, D and E subtypes of α1 subunit. The result showed us a possibility that phenotype of voltage-gated calcium channel α1 subunit were not depend on cell kind.Labeling neuroepithelium in neural tube of chicken embryo or immunohistochemical staining of anti HNK-1 antibody showed migrating neural crest cells in the mesodermal region. S phase of neural crest cells were detected by immunohistochemical staining of BrdU applied to chicken embryo before fixation. 1) Migrating neural crest cells in mesodermal region proliferated on their way from neural crest. Immunohistochemical staining A, B, C, D and E subtypes of voltage-gated calcium channel α1 subunit in mouse embryonic day 8 to day 9.5 were detected by immunohistochemical staining. 2) C and D types of α1 subunit among 5 types were strongly expressed in the beginning of neural crest cells migration. 3) During their migration, C, D and E types were expressed. 4) No α1 subunit were expressed in neuroepithelium of neural tube at embryonic day 8, all type of α1 subunits were scattered at embryonic day 9.5. These results suggested that the voltage-gated calcium channels are involved in neural crest cell's migration from the beginning of and during migration. Not secretion nor neurotransmission, novel function of voltage-gated calcium channel on neuroepithelium is suggested on account of 4).