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. 2000 Mar;46(3):370–375. doi: 10.1136/gut.46.3.370

Pathways and receptors involved in peptide YY induced contraction of rat proximal colonic muscle in vitro

L Ferrier 1, J Segain 1, P Pacaud 1, C Cherbut 1, G Loirand 1, J Galmiche 1, H Blottiere 1
PMCID: PMC1727869  PMID: 10673299

Abstract

BACKGROUND—Peptide YY (PYY) is involved in the regulation of several gut functions, including secretion and motility. It exerts its effects through a family of six receptors, commonly named the Y receptor family.
AIMS—To characterise the effects of PYY on strips of rat proximal colon in vitro, and to determine the pathways and receptors involved.
METHODS—Contractions of strips removed from the muscle layer of rat proximal colon were recorded under isometric conditions, using PYY, Y receptor agonists and antagonists, and nerve blockers. Reverse transcription-polymerase chain reaction was also performed to detect the presence of mRNA coding for Y receptors. Finally, smooth muscle cells were isolated to estimate the cell length and intracellular Ca2+ concentration in the presence and absence of PYY.
RESULTS—PYY, neuropeptide Y (NPY), pancreatic polypeptide (PP) and [Leu31,Pro34]NPY induced a dose dependent contraction of strips from proximal colon. Tetrodotoxin partially inhibited the PYY and NPY induced contractions, and strongly inhibited the PP induced contraction. Specific antagonists showed the involvement of cholinergic nicotinic receptors and NK1 receptor. BIBP 3226, a specific Y1 antagonist, did not modify the colonic smooth muscle response to PYY, whereas blocking L-type Ca2+ channels with D-600 abolished its effects. Moreover, PYY induced an increase in intracellular Ca2+ concentration, associated with a reduction in cell length. mRNA encoding Y1 and Y4 receptors were detected in the muscle strips.
CONCLUSIONS—These findings suggest that PYY stimulates colonic contractile activity in vitro through (a) a nervous Y4 dependent pathway and (b) a pathway involving a potential new receptor on myocytes.


Keywords: peptide YY; Y receptors; colon; motility; myocytes

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Figure 1  .

Figure 1  

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(A) Contraction elicited by 10−5 M peptide YY (PYY) compared with 10−6 M acetylcholine chloride (ACh) on the same colon nerve/muscle preparation. (B) Dose-response curve representing the effects of PYY on the proximal colon nerve/muscle preparation. Results are mean (SEM), n = 7.

Figure 2  .

Figure 2  

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Effect of 10−6 M acetylcholine chloride (ACh) and 10−6 M peptide YY (PYY) on the length of isolated colonic smooth muscle cells. This is a representative of the three experiments performed. Results are expressed as mean (SEM) (n = 60 cells). **p<0.01, ***p<0.001 compared with control.

Figure 3  .

Figure 3  

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Increase in intracellular Ca2+ concentration in isolated smooth muscle cells after exposure to 10−5 M peptide YY (PYY) or 10−5 M acetylcholine chloride (ACh), as observed using Indo-1 fluorescent Ca2+ dye.

Figure 4  .

Figure 4  

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Effect of neuropeptide Y (NPY)(13-36) and [Leu31,Pro34]NPY on the proximal colon nerve/muscle preparation (NMP). Results are expressed as mean (SEM), n = 7. ACh, acetylcholine chloride.

Figure 5  .

Figure 5  

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(A) Comparison of contractions in the proximal colon nerve/muscle preparation (NMP) induced by peptide YY (PYY) and pancreatic peptide (PP) before and after tetrodotoxin 10−6 M application. (B) Comparison of contractions in the proximal colon nerve/muscle preparation induced by PYY and neuropeptide Y (NPY) before and after tetrodotoxin 10−6 M application. ACh, acetylcholine chloride. TTX, tetrodotoxin. *p<0.05, **p<0.01 compared with before tetrodotoxin application.

Figure 6  .

Figure 6  

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RT-PCR performed on nerve/muscle strips from proximal colon for Y1, Y2, Y4, and Y5 receptors. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is the housekeeping gene.

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