Keratin binding to 14-3-3 proteins modulates keratin filaments and hepatocyte mitotic progression.

Keratin polypeptides 8 and 18 (K8/18) are the major intermediate filament proteins of simple-type epithelia. K18 Ser-33 phosphorylation regulates its binding to 14-3-3 proteins during mitosis. We studied the significance of keratin binding to 14-3-3 in transgenic mice that overexpress wild-type or Ser-33-->Ala (S33A) K18. In S33A but not wild-type K18-overexpressing mice, pancreatic acinar cell keratin filaments retracted from the basal nuclear region and became apically concentrated. In contrast, K18 S33A had a minimal effect on hepatocyte keratin filament organization. Partial hepatectomy of K18-S33A-overexpressing mice did not affect liver regeneration but caused limited mitotic arrest, accumulation of abnormal mitotic figures, dramatic fragmentation of hepatocyte keratin filaments, with retention of a speckled 14-3-3zeta mitotic cell nuclear-staining pattern that usually becomes diffuse during mitosis. Hence, K18 Ser-33 phosphorylation regulates keratin filament organization in simple-type epithelia in vivo. Keratin binding to 14-3-3 may partially modulate hepatocyte mitotic progression, in association with nuclear redistribution of 14-3-3 proteins during mitosis.

A Method for Immobilizing the Forelimbs of Rabbits.

Immobilizing the forelimbs of rabbits after surgical procedures is necessary to allow healing, yet it often can be difficult, because rabbits are often able to pull the repaired limb from its cast soon after surgery and well before adequate tissue repair has taken place. We describe here a method of immobilization that uses 3 layers of cast material combined with flexion of the radiocarpal and radiohumeral joints. This method resulted in successful immobilization in 97% of the rabbits on which it was used.