Rodríguez-Martín E, Boyano-Adánez M C, Bodega G, Martín M, Hernández C, Quin Y, Vadillo M, Arilla-Ferreiro E
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Alcalá, Madrid, Spain.
FEBS Lett. 1999 Feb 26;445(2-3):356-60. doi: 10.1016/s0014-5793(99)00133-7.
Freshly enzymatically isolated pancreatic acini from lactating and weaning Wistar rats were used to investigate the role of protein kinase C (PKC) isoforms during these physiologically relevant pancreatic secretory and growth processes. The combination of immunoblot and immunohistochemical analysis shows that the PKC isoforms alpha, delta, and epsilon are present in pancreatic acini from control, lactating and weaning rats. A vesicular distribution of PKC-alpha, -delta, and -epsilon was detected by immunohistochemical analysis in the pancreatic acini from all the experimental groups. PKC-delta showed the strongest PKC immunoreactivity (PKC-IR). In this vesicular distribution, PKC-IR was located at the apical region of the acinar cells. No differences were observed between control, lactating and weaning rats. However, the immunoblot analysis of pancreatic PKC isoforms during lactation and weaning showed a significant translocation of PKC-delta from the cytosol to the membrane fraction when compared with control animals. Translocation of PKC isoforms (alpha, delta and epsilon) in response to 12-O-tetradecanoyl phorbol 13-acetate (TPA) 1 microM (15 min, 37 degrees C) was comparable in pancreatic acini from control, lactating and weaning rats. In the control group, a significant translocation of all the isoforms (alpha, delta and epsilon) from the cytosol to the membrane was observed. The PKC isoform most translocated by TPA was PKC-delta. In contrast, no statistically significant increase in PKC-delta translocation was detected in pancreatic acini isolated from lactating or weaning rats. These results suggest that the PKC isoforms are already translocated to the surface of the acinar cells from lactating or weaning rats. In addition, they suggest that isoform specific spatial PKC distribution and translocation occur in association with the growth response previously described in the rat exocrine pancreas during lactation and weaning.
从哺乳期和断奶期的Wistar大鼠新鲜酶解分离的胰腺腺泡,被用于研究蛋白激酶C(PKC)亚型在这些生理相关的胰腺分泌和生长过程中的作用。免疫印迹和免疫组织化学分析相结合表明,PKC亚型α、δ和ε存在于对照、哺乳期和断奶期大鼠的胰腺腺泡中。通过免疫组织化学分析在所有实验组的胰腺腺泡中检测到PKC-α、-δ和-ε的囊泡分布。PKC-δ显示出最强的PKC免疫反应性(PKC-IR)。在这种囊泡分布中,PKC-IR位于腺泡细胞的顶端区域。对照、哺乳期和断奶期大鼠之间未观察到差异。然而,与对照动物相比,哺乳期和断奶期胰腺PKC亚型的免疫印迹分析显示PKC-δ从细胞质显著易位到膜部分。对照、哺乳期和断奶期大鼠胰腺腺泡中,1 μM 12-O-十四酰佛波醇13-乙酸酯(TPA)(15分钟,37℃)诱导的PKC亚型(α、δ和ε)易位情况相当。在对照组中,观察到所有亚型(α、δ和ε)从细胞质到膜的显著易位。TPA诱导易位最多的PKC亚型是PKC-δ。相比之下,从哺乳期或断奶期大鼠分离的胰腺腺泡中,未检测到PKC-δ易位有统计学意义的增加。这些结果表明,PKC亚型已从哺乳期或断奶期大鼠的腺泡细胞易位到表面。此外,它们表明,在哺乳期和断奶期大鼠外分泌胰腺中,亚型特异性的PKC空间分布和易位与先前描述的生长反应相关。