Upregulation of molecular motor-encoding genes during hepatocyte growth factor-and epidermal growth factor-induced cell motility

Torok, Natalia J; Urrutia, Raúl; Nakamura, Toshikazu; McNiven, Mark A.

doi:10.1002/(sici)1097-4652(199606)167:3<422::aid-jcp6>3.0.co;2-r

Cited by 21 publications

(1 citation statement)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Western blotting procedure is performed as described previously (Henley and McNiven, 1996) except using 10% nonfat dry milk as a blocking reagent. Northern blot analysis was essentially carried out using the methods described in a previous publication (Török et al, 1996).…”

Section: Sds-page and Western And Northern Blot Analysesmentioning

confidence: 99%

A Novel Dynamin-like Protein Associates with Cytoplasmic Vesicles and Tubules of the Endoplasmic Reticulum in Mammalian Cells

Yoon

Pitts

Dahan

et al. 1998

The Journal of Cell Biology

Self Cite

170

192

View full text Add to dashboard Cite

Abstract. Dynamins are 100-kilodalton guanosine triphosphatases that participate in the formation of nascent vesicles during endocytosis. Here, we have tested if novel dynamin-like proteins are expressed in mammalian cells to support vesicle trafficking processes at cytoplasmic sites distinct from the plasma membrane. Immunological and molecular biological methods were used to isolate a cDNA clone encoding an 80-kilodalton novel dynamin-like protein, DLP1, that shares up to 42% homology with other dynamin-related proteins. DLP1 is expressed in all tissues examined and contains two alternatively spliced regions that are differentially expressed in a tissue-specific manner. DLP1 is enriched in subcellular membrane fractions of cytoplasmic vesicles and endoplasmic reticulum. Morphological studies of DLP1 in cultured cells using either a specific antibody or an expressed green fluorescent protein (GFP)- DLP1 fusion protein revealed that DLP1 associates with punctate cytoplasmic vesicles that do not colocalize with conventional dynamin, clathrin, or endocytic ligands. Remarkably, DLP1-positive structures coalign with microtubules and, most strikingly, with endoplasmic reticulum tubules as verified by double labeling with antibodies to calnexin and Rab1 as well as by immunoelectron microscopy. These observations provide the first evidence that a novel dynamin-like protein is expressed in mammalian cells where it associates with a secretory, rather than endocytic membrane compartment.

show abstract

Section: Sds-page and Western And Northern Blot Analysesmentioning

confidence: 99%

A Novel Dynamin-like Protein Associates with Cytoplasmic Vesicles and Tubules of the Endoplasmic Reticulum in Mammalian Cells

Yoon

Pitts

Dahan

et al. 1998

The Journal of Cell Biology

Self Cite

170

192

View full text Add to dashboard Cite

show abstract

Differential modulation of hepatocyte growth factor-stimulated motility by transforming growth factor β1 on rat liver epithelial cells in vitro

Stolz

Michalopoulos

1998

J. Cell. Physiol.

View full text Add to dashboard Cite

We have previously shown that transforming growth factor-beta1 (TGF-beta1) enhances the epidermal growth factor- (EGF) and transforming growth factor-alpha (TGF-alpha)-stimulated motility of rat hepatocytes in an extracellular matrix (ECM)-dependent fashion (Stolz and Michalopoulos, 1997, J. Cell. Physiol., 170:57-68). We have extended this study to examine the effects of TGF-beta1 on hepatocyte growth factor (HGF) and EGF-stimulated motility of rat nonparenchymal liver epithelial cells (RLECs) in vitro and determined that chemotaxis, scattering, and monolayer wound healing by EGF was synergistically enhanced by TGF-beta1 on all ECMs examined. However, HGF-based motility, unlike EGF-stimulated motility, was modulated in an assay-dependent manner by TGF-beta1. HGF-stimulated chemotaxis was dramatically decreased by addition of TGF-beta1, but wound healing was synergistically enhanced by TGF-beta1 on all ECMs examined. HGF-based scattering was not consistently affected by TGF-beta1 on any ECM tested except on laminin, where scattering was often reduced by the concomitant addition of TGF-beta1. TGF-beta1 enhanced the motility associated with monolayer wound healing by HGF or EGF independent of DNA synthesis, because tritiated thymidine uptake was consistently reduced by 60% in the presence of TGF-beta1. The data indicate that HGF and EGF motility do not follow redundant signal-transduction pathways and that specific growth factor motility-related events, as measured by wound healing, scattering, and chemotaxis, are modulated independently by ECM and TGF-beta1.

show abstract

Mechanochemical manipulation of hepatocyte aggregation can selectively induce or repress liver-specific function

Semler

Ranucci

Moghe

2000

Biotechnol. Bioeng.

118

View full text Add to dashboard Cite

Controlled activation of hepatocyte aggregation is critical to three‐dimensional (3D) multicellular morphogenesis during native regeneration of liver as well as tissue reconstruction therapies. In this work, we quantify the stimulatory effects of two model hepatotrophic activators, epidermal growth factor (EGF) and hepatocyte growth factor (HGF), on the aggregation kinetics and liver‐specific function of hepatocytes cultured on organotypic substrates with differing mechanical resistivity. Substrate‐specific morphogenesis of cultured hepatocytes is induced on a tissue basement membrane extract, Matrigel, formulated at two distinct levels of mechanical compliance (storage modulus G′, at oscillatory shear rate 1 rad/s, was 34 Pa for basal Matrigel and 118 Pa for crosslinked Matrigel). Overall, we report that growth factor stimulation selectively promotes the kinetics of aggregation in the form of two‐dimensional corded aggregates on basal Matrigel and three‐dimensional spheroidal aggregates on crosslinked Matrigel. Our analysis also indicates that costimulation with EGF and HGF (20 ng/mL each) cooperatively maximizes the kinetics of aggregation in a substrate‐specific manner. In addition, we show that the role of growth factor stimulation on hepatocyte function is sensitively governed by the mechanical compliance of the substrate. In particular, on matrices with high compliance, costimulatory aggregation is shown to elicit a marked increase in albumin secretion rate, whereas on matrices with low compliance aggregation results in effective functional repression to basal, unstimulated levels. Thus, our studies highlight a novel interplay of physicochemical parameters of the culture microenvironment, leading to selective enhancement or repression of differentiated functions of hepatocytes, in concert with the activation of cellular morphogenesis. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 69: 359–369, 2000.

show abstract

Upregulation of molecular motor-encoding genes during hepatocyte growth factor-and epidermal growth factor-induced cell motility

Cited by 21 publications

References 57 publications

A Novel Dynamin-like Protein Associates with Cytoplasmic Vesicles and Tubules of the Endoplasmic Reticulum in Mammalian Cells

A Novel Dynamin-like Protein Associates with Cytoplasmic Vesicles and Tubules of the Endoplasmic Reticulum in Mammalian Cells

Differential modulation of hepatocyte growth factor-stimulated motility by transforming growth factor β1 on rat liver epithelial cells in vitro

Mechanochemical manipulation of hepatocyte aggregation can selectively induce or repress liver-specific function

Contact Info

Product

Resources

About