Evidence for Direct Involvement of β‐Catenin in Phorbol Ester‐Induced Neurite Outgrowth in GT1‐1 Hypothalamic Neurones

Abstract

Gonadotropin-releasing hormone (GnRH) is a pivotal neuroendocrine regulator controlling reproductive functions. However, the scattered distribution of GnRH neurones in the mammalian brain has hindered studies on the development and differentiation of GnRH neurones. In the present study, we used the immortalized GnRH-producing GT1-1 cells to examine whether activation of protein kinase C (PKC) pathway with 12-O-tetradecanoyl-13-acetate (TPA) induces morphological and functional differentiation of GnRH neurones. TPA induced neurite outgrowth and inhibited proliferation of GT1-1 cells that were specifically antagonized by cotreatment of PKC inhibitor, calphostin C. The functional significance of TPA-induced differentiation of GT1-1 cells was manifested in part by the changes in the effects of gamma-aminobutyric acid (GABA) on intracellular Ca2+ levels. In untreated GT1-1 cells, activation of GABA-A receptor with 10 microM muscimol increased intracellular Ca2+ levels, whereas such stimulatory effects disappeared in GT1-1 cells bearing neurites. Accordingly, muscimol could not stimulate GnRH release in TPA-treated GT1-1 cells. To elucidate the molecular mechanism underlying TPA-induced neurite outgrowth, we performed differential display reverse transcription-polymerase chain reaction. Among several genes that are affected by TPA treatment, we found a significant induction of beta-catenin mRNA expression. Along with the rapid induction of beta-catenin protein levels, we observed that beta-catenin was reallocated from cell-cell adhesion sites to the growth cones within 3 h of TPA treatment. Transient transfection studies with green fluorescent protein as a reporter gene demonstrated that beta-catenin overexpression alone can promote neurite outgrowth in GT1-1 cells. Moreover, TPA was found to increase the transcription-activational roles of beta-catenin. Together, these data provide evidence that beta-catenin is involved in the TPA-induced functional differentiation of immortalized GnRH neurones.

Cell Differentiation of Gonadotropin-Releasing Hormone Neurons and Alternative RNA Splicing of the Gonadotropin-Releasing Hormone Transcript

“…This excitatory GABAergic action on intracellular Ca 2+ was abolished when GT1 cells were pretreated with TPA for 48 h [23]. This result correlates well with the decrease in GnRH secretion.…”

“…It is possible that the stimulation of GnRH secretion by TPA is due to a transient action on the releasable pool, which may be independent of subsequent biosynthesis of the decapeptide. Recently, we found that TPA promoted morphological differentiation of GnRH neurons [23]. TPA induced NF200 and GAP-43 expression along with extension of neurites.…”

“…Induction of tyrosine-phosphorylated β-catenin by TPA treatment reduced cadherin-mediated cell adhesion as well as the rearrangement of the actin cytoskeleton, inducing an increase in cellular motility and neurite outgrowth [23]. In figure 1, we show our working hypothesis of PKC-induced neurite outgrowth of GnRH neurons.…”

Glucocorticoid inhibits growth factor‐induced differentiation of hippocampal progenitor HiB5 cells

“…Vehicle (0.1% ethanol), DEX (1 n m to 1 µ m ) and/or RU486 (5 µ m ) were treated 24 h prior to PDGF treatment. Neurite outgrowth was counted as previously described (Sun et al . 2001).…”

Activated Hair Follicle Stem Cells and Wnt/β-catenin Signaling Involve in Pathnogenesis of Sebaceous Neoplasms

“…Trempus et al reported that up-regulated β-catenin was involved in the initiation and development of papilloma induced by DMBA/TPA by maintaining the activity of CD34-positive HFSCs 19. Furthermore, β-catenin was up-regulated in cultured GT1-1 neuronal cells treated by TPA 25. These results indicated that Wnt10b/β-catenin signaling is activated by continuous TPA treatment in a small population of basal sebaceous gland cells.…”