骨髓基质细胞和CD34+的造血祖细胞间的交流由connexin43型通道连接调节

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Intercellular communication between bone marrow stromal cells and CD34+ haematopoietic progenitor cells is mediated by connexin 43-type gap junctions
. D3 ^  ~* o$ o! X( g7 A骨髓基质细胞和CD34+的造血祖细胞间的交流由connexin43型通道连接调节- C) i9 ^. q, e& x& d! ?5 E
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摘要：
  C  h# ^- i3 c/ s. y0 R5 h8 R8 u关于造血祖细胞（HPC）和基质细胞间是否存在功能性的通道连接，一直存在着争议。原始的CD34+的HPC与人类骨髓基质细胞系L87/4共培养。利用具有高度敏感性的全细胞钳技术，可以发现大多数的（91%）CD34+的HPC与L87/4间有电流交流。更重要的是，）CD34+的HPC依附在L87/4后的1小时内可以观察到有效的偶联。通过对比，由L87/4组成的同类细胞间有显著更高的电偶联。单通道导电率分析发现了一种形式，以connexin43型通道连接为特征，对于同类和不同类的细胞都是这样的。这个Cx表型通过Cx-43特异性的单克隆抗体流式细胞仪分析和RT-PCR的验证。最后，染料转染实验利用降落伞技术使得染料扩散而不影响浆膜成分，这个实验是对电生理实验的补充。总的来说，我们的数据表明，功能性的Cx-43型通道连接存在于基质细胞和未成熟的造血祖细胞中，可能提供造血过程中一个重要的调节通道。" S9 ]  W! t& X  p5 x; C' k
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Summary. The existence of functional gap junctions between haematopoietic progenitor cells (HPCs) and stromal
6 o" N" Z, q3 J2 k  ]cells of the haematopoietic microenvironment in the human system is a controversial issue. Primary CD341 HPCs
# c" n& G, |) ~4 c7 y% @& n, ^8 D' I" ~isolated from leukapheresis products were co-incubated with the human fibroblastoid bone marrow stromal cell
5 @" ^0 `; Z4 z$ t' U: ?( {line L87/4 in short-term liquid culture. Using the highly sensitive double whole-cell patch-clamp technique, we$ w5 Z+ h5 g+ Y6 `# E) S/ u
found that the majority (91%) of CD341 HPCs are electrically coupled to L87/4 cells. Importantly, efficient
/ ?" m0 Q$ K+ I$ Y5 L5 ucoupling was observed within 1 h of the attachment ofCD341 HPCs to plastic adherent L87/4 cells. By comparison,
( b0 E& H" }. `: l+ C9 L% h5 n2 khomologous cell pairs formed by L87/4 cells exhibited a significantly higher electric coupling. Analysis of singlechannel conductances revealed an electric profile characteristic of connexin 43 (Cx43)-type gap junctions for both homologous and heterologous cell pairs. The Cx phenotype was confirmed using Cx43-specific monoclonal antibodies in a flow cytometric assay and reverse transcription polymerase chain reaction (RT-PCR) for the detection of Cx43 mRNA. Finally, the electrophysiological studies were complemented by dye-transfer experiments using the recently described `parachute' technique that allows the monitoring of dye diffusion without disruption of the plasma membrane. Taken together, our data indicate that functional Cx43-type gap junctions exist between stromal cells and immature HPCs and, thus, may provide an important regulatory pathway in haematopoiesis.& ~6 I- `) G0 F% |- Y

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