Kita M, Tanaka K, Shinmura K, Tanaka Y, Liu Y, Imanishi J
Départements de Microbiologie et d'Ophtalmologie, Université Médicale de la Préfecture de Kyoto, Japan.
C R Seances Soc Biol Fil. 1994;188(5-6):593-600.
The expression of the cytokine and IFN-related genes was studied in mouse embryo using RT-PCR method capable of detecting low levels of mRNA. Total RNA was prepared from the days 7 embryos by acid guanidinium thiocyanate-phenol-chloroform (AGPC) method. cDNA was synthesized by M-MLV reverse transcriptase, and amplified using the specific oligonucleotide primers for IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, TNF-alpha, IFN-alpha, IFN-beta, IFN-gamma, IFN-alpha/beta receptor (IFN-alpha/beta R), IFN-gamma receptor (IFN-gamma R), interferon reguratory factor (IRF)-1, IRF-2 and 2'-5' oligoadenylate synthetase (2-5AS) by PCR method. Although the expressions of IL-1 beta, IL-4, IL-5, IL-6, TNF-alpha and IFN-gamma mRNA were detected in all the embryos tested, the expressions of IL-2, IL-3, IFN-alpha and IFN-beta mRNA were not detected at all. On the other hand, the expressions of IFN-related genes such as IFN-alpha/beta R, IFN-gamma R, IRF-1, IRF-2 and 2-5AS mRNA, were also detected. These results suggest that these cytokine may play an important role in early embryonic development.
采用能够检测低水平mRNA的逆转录聚合酶链反应（RT-PCR）方法，对小鼠胚胎中细胞因子和干扰素相关基因的表达进行了研究。通过酸性异硫氰酸胍-苯酚-氯仿（AGPC）法从7日龄胚胎中制备总RNA。用M-MLV逆转录酶合成cDNA，并使用针对白细胞介素-1（IL-1）、白细胞介素-2（IL-2）、白细胞介素-3（IL-3）、白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、干扰素-α（IFN-α）、干扰素-β（IFN-β）、干扰素-γ（IFN-γ）、干扰素-α/β受体（IFN-α/βR）、干扰素-γ受体（IFN-γR）、干扰素调节因子（IRF）-1、IRF-2和2'-5'寡腺苷酸合成酶（2-5AS）的特异性寡核苷酸引物，通过聚合酶链反应（PCR）方法进行扩增。虽然在所有检测的胚胎中均检测到IL-1β、IL-4、IL-5、IL-6、TNF-α和IFN-γmRNA的表达，但未检测到IL-2、IL-3、IFN-α和IFN-βmRNA的表达。另一方面，也检测到了IFN相关基因如IFN-α/βR、IFN-γR、IRF-1、IRF-2和2-5AS mRNA的表达。这些结果表明，这些细胞因子可能在早期胚胎发育中发挥重要作用。
