Peptide YY and Neuropeptide Y Induce Villin Expression, Reduce Adhesion, and Enhance Migration in Small Intestinal Cells through the Regulation of CD63, Matrix Metalloproteinase-3, and Cdc42 Activity

Abstract

Peptide YY (PYY) and neuropeptide Y (NPY) are regulatory peptides synthesized in the intestine and brain, respectively, that modify physiological functions affecting nutrient assimilation and feeding behavior. Because PYY and NPY also alter the expression of intestine-specific differentiation marker proteins and the tetraspanin CD63, which is involved in cell adhesion, we investigated whether intestinal cell differentiation could be linked to mucosal cell adhesion and migration through these peptides. PYY and NPY significantly decreased cell adhesion and increased cell migration in a dose-dependent manner prior to cell confluency in our model system, non-tumorigenic small intestinal hBRIE 380i cells. Both peptides reduced CD63 expression and CD63-dependent cell adhesion. CD63 overexpression increased and antisense CD63 cDNA decreased intestinal cell adhesion. In parallel, both PYY and NPY increased expression of matrix metalloproteinase-3 (MMP-3) to a level sufficient to induce cell migration by activating the Rho GTPase Cdc42. The effects of both peptides on cell migration were blocked in cells constitutively overexpressing dominant-negative Cdc42. PYY and NPY also significantly induced the expression of the differentiation marker villin, which could be eliminated by an MMP inhibitor at a concentration that inhibits cell migration. Increased MMP-3 activity, which enhanced cell migration, also induced villin mRNA levels. Therefore, these data indicate that the alteration of adhesion and migration by PYY and NPY occurs in part by synchronous modulation of three proteins that are involved in extracellular matrix-basolateral membrane interactions, CD63, MMP-3 and Cdc42, and that PYY/NPY regulation of expression of mucosal proteins such as villin is linked to the process of cell migration and adhesion.

The tetraspanin superfamily member NET-6 is a new tumor suppressor gene

“…Tetraspanins also have the ability to interact directly or indirectly with a diverse array of proteins, such as integrins, immunoreceptors and other molecules which play a critical role in cell signaling and apoptosis (Cherukuri et al 2004;Chattopadhyay et al 2003;Carloni et al 2004;Zhang et al 2001;Berditchevski et al 2002;Murayama et al 2004;Shinohara et al 2001). Some of the best studied tetraspanins including CD9, CD63 and CD82 (KAI1), have been found to function as tumor suppressor genes (Murayama et al 2004;Shinohara et al 2001;Huang et al 2004;Dong et al 1995;Furuya et al 2005;Lee et al 2005;Jang and Lee. 2003).…”

Feeding-dependent activation of enteric cells and sensory neurons by lymphatic fluid: evidence for a neurolymphocrine system

“…Reverse transcription was performed as previously described (47). The PCR primers for LPA5 were designed to match the LPA5 sequence of rat (accession number: XM_575667) and mouse (accession number: BC117528).…”

Identification of a protein hydrolysate responsive G protein-coupled receptor in enterocytes

“…Reverse transcription was as described previously (40). PCR was carried out using Taq DNA polymerase (New England Biolab).…”

“…PCR parameters for GPR93 were as follows: 20 s at 94°C, 15 s at 55°C, and 30 s at 72°C for 32 cycles. The primers and PCR condition for villin, I-FABP, and 18S RNA were as previously described (40). Amplified cDNA fragments were analyzed by agarose gel electrophoresis followed by densitometry.…”