Morphological plasticity and rearrangement of cytoskeletons in pituicytes cultured from adult rat neurohypophysis.

Abstract

The adult rat neurohypophysis reveals drastic morphological plasticity of neuron-glial organization during chronic physiological stimulation. Pituicytes are modified astrocytes in the neurohypophysis, and shape conversion of them largely contributes to the morphological plasticity. The present study aimed to investigate the receptor-mediated mechanism for shape conversion of the pituicyte morphology, particularly in relation with changes of cytoskeletal organization. The cultured pituicytes from adult rat neurohypophysis were mostly flat amorphous shape in normal salt solution. Histochemical experiments showed that thick bundle of microfilament (stress fibers) and fine fibers of microtubule distributed evenly within the pituicyte. When pituicytes were treated with adenosine (more than 1 microM), isoproterenol (IPR); beta-agonist, more than 10 nM), and dibutyryl cyclic AMP (dBcAMP, 1 mM), the pituicyte morphology changed from flat to stellate shape. Upon treatment with dBcAMP, stress fibers within pituicyte cytoplasm disappeared, and microtubule assembled in the cellular processes and cytoplasm surrounding the nucleus. Pretreatment with colchicine (microtubule-disrupting agent, 25 microM) and orthovanadate (tyrosine phosphatase inhibitor, 1 mM) prevented dBcAMP-induced stellation of the pituicyte morphology. Treatment with sphingosine (protein kinase C inhibitor, 10 microM), W-7 (calmodulin dependent protein kinase inhibitor, 40 microM), ML-9 (myosin light chain kinase inhibitor, 20 microM), and cytochalasinB (CytB; microfilament disrupting agent, 5 microM), induced stellation of the pituicyte morphology. Treatment of endothelin-1 (more than 0.1 nM) and endotheline-3 (more than 0.1 nM) reverted dBcAMP-induced stellation of the pituicyte morphology to original flat one and also reverted arrangement of cytoskeletons of stress fiber and microtubules as seen in control one. The present results reveal that pituicyte shape conversion is mediated via beta-adrenergic, adenosine and endotheline and depend on rearrangement of stress fibers and microtubules. In addition, the mechanism of shape conversion of pituicytes cultured from adult neurohypophysis is quite similar to that of astrocytes cultured from neonatal brains and possibly is useful for understanding morphological plasticity of adult brains.