Kyoko Furukawa

Effects of Thyrotropin-Releasing Hormone (TRH) on the isolated small intestine and taenia coli of the guinea pig

Thyrotropin-releasing hormone (TRH) produced a contraction in the isolated segment of duodenum and taenia coli of the guinea-pig (pA2, 8.0 and 8.9). TRH induced a contraction, a relaxation, or a contraction followed by relaxation in the jejunum and ileum. All the responses to TRH of the small intestine and the taenia coli were abolished in the presence of tetrodotoxin but not affected by hexamethonium. The contractile response to TRH of the small intestine was abolished and replaced by a relaxation in the presence of hyoscine. This relaxant response was not affected by guanethidine. The taenial response to TRH was partially inhibited by either hyoscine or methysergide and markedly diminished by the two together. These findings indicate that TRH acts on the myenteric neurons of the small intestine and taenia coli of the guinea pig. The contractile response of small intestine is likely to be induced through cholinergic nerves while cholinergic, serotonergic and unidentified excitatory neurons seem to be involved in the taenial response. These neurogenic actions of TRH on the guinea-pig intestine are in contrast with the myogenic natur of the response to TRH in the duodenum of the rat.

Postnatal changes in response to adenosine and adenine nucleotides in rat duodenum.

1 The effects of adenosine and adenine nucleotides were studied in rat duodenum from postnatal day 1 to day 70. The mechanical activity of duodenal segments was recorded through an isotonic transducer connected to a polygraphic recorder. 2 In rat duodenal segments, adenosine‐5′‐triphosphate (ATP, 10−4m) and adenosines‐5′‐diphosphate (ADP, 10−4m) produced a contractile response on postnatal day 1. This response increased with age, peaking on day 7, followed by a gradual decrease and was non‐existent by day 21. In contrast, a relaxant response to ATP and ADP was apparent on day 21, and continued to increase up to day 70. 3 The contraction caused by ATP was inhibited by indomethacin or the P2y‐purinoceptor antagonist, reactive blue‐2 but not by tetrodotoxin or hyoscine. Thus, it may be mediated by production of prostaglandin through the P2y‐purinoceptor. The relaxation produced by ATP was inhibited by reactive blue‐2 but not by tetrodotoxin, guanethidine or the P1‐purinoceptor antagonist, 8‐phenyltheophylline indicating that ATP acts on smooth muscle directly through the P2y‐purinoceptor. The pD2 for the contractile response to ATP was 5.15 on day 7 and that for the relaxant response, 6.64 on day 70. 4 Adenosine (10−4m) and adenosine‐5′‐monophosphate (AMP, 10−4m) elicited no response before day 14. On day 14, both adenosine and AMP produced a small relaxant response which increased with age. The relaxation produced by adenosine was inhibited by 8‐phenyltheophylline but not by tetrodotoxin or guanethidine, indicating that it is mediated by an action on the P1‐purinoceptor of smooth muscle. 5 It is evident from these results that in neonatal rat, a contractile response to ATP and ADP occurs initially in the duodenum, followed by a relaxant response to adenosine and AMP on day 14 and to ADP and ATP on day 21. 6 The smooth muscle of rat duodenum may tentatively be concluded to contain separate purinoceptors for adenosine and AMP (P1) and ADP and ATP (P2) and the responses to P1‐ and P2‐agonists change during the course of development.

Developmental changes in the response of rat isolated duodenum to nicotine

Developmental changes in the response to ganglionic stimulants, nicotine and dimethylphenylpiperazinium, were investigated in rat isolated duodenum by recording isotonic mechanical activity. The duodenal response to nicotine/dimethylphenylpiperazinium (3 x 10(-7) to 10(-3) M) in neonatal rats was contraction, which was blocked by hexamethonium, tetrodotoxin and hyoscine. The response to nicotine/dimethylphenylpiperazinium (10(-6) to 10(-4) M) in the adult duodenum was relaxation, which was blocked by tetrodotoxin and hexamethonium, but by neither guanethidine nor hyoscine. The transition of the response to nicotine/dimethylphenylpiperazinium from contraction to relaxation occurred at around the 3rd postnatal week. Nicotine-induced relaxation of adult duodenum was significantly inhibited by preincubation with alpha-chymotrypsin, a proteolytic enzyme, and a combination of nucleotide pyrophosphatase and 8-phenyltheophylline, a P1 purinoceptor antagonist. Nicotine-induced relaxation was desensitized by alpha, beta-methylene ATP, a stable P2x purinoceptor agonist. These results suggest that the contractile response of isolated duodenum to nicotine is mediated through cholinergic transmission in neonatal rats and the relaxant response is mediated through non-adrenergic, non-cholinergic transmission, which involves both peptidergic and purinergic transmission, in adult rats.

Myogenic receptivity for Met-enkephalin in the rat duodenum: Comparison with the guinea-pig duodenum

Met-enkephalin (ENK) induced a dose-dependent transient relaxation in vitro in the rat duodenum while morphine neither induced a response nor affected the response to ENK. The response was not blocked by tetrodotoxin but was abolished by naloxone. The response of guinea-pig duodenum to ENK was either relaxation or pulsatile contractions depending on the dose and was abolished by tetrodotoxin.

Postnatal change in receptivity for methionine5-enkephalin in rat duodenum: Transition from neurogenic to myogenic receptivity

The study concerned the postnatal ontogenesis of the response to methionine5-enkephalin (ENK) in the duodenum of developing rats. Sustained relaxation was produced by ENK in the isolated duodenum of 8 day old rats. The response was blocked by tetrodotoxin or naloxone. The response increased by day 18, decreased thereafter and was extinguished after 40 days. On the other hand, a tetrodotoxin resistant response to ENK, transient relaxation, appeared at day 20 and was augmented thereafter. The development of neurogenic receptivity for ENK preceded the appearance of myogenic receptivity in the rat duodenum.

Contractile response to neuropeptide Y of rat isolated duodenum.

The response of isolated duodenum to neuropeptide Y (NPY) was studied isotonically in neonatal and adult rats. Neuropeptide Y (10(-8) to 10(-6) M) elicited a biphasic contraction of isolated duodenum from neonatal rats, but monophasic and weak contraction of adult duodenum. The first phase of NPY-induced contraction of neonatal duodenum was concentration dependent and partially inhibited by preincubation with tetrodotoxin, a Na+ channel blocker, hyoscine, a muscarinic antagonist, suramin, a P2 purinoceptor antagonist, and indomethacin, an inhibitor for prostaglandin biosynthesis. Neuropeptide Y(13-36), a specific Y2 NPY receptor agonist, elicited a concentration-dependent contraction of neonatal rat duodenum. The duodenal response to NPY thus changes during development in rats. Both cholinergic and purinergic transmission and prostaglandin biosynthesis may be involved in the NPY-induced contraction of neonatal duodenum. Neuropeptide Y-induced contraction may be mediated through presynaptic Y2 receptors.

Effects of pGlu-His-Pro-amphetamine (TRH-amphetamine) on the isolated duodenum of the guinea-pig: antagonistic effect of amphetamine on TRH response.

pGlu-His-Pro-dexamphetamine (TRH-A) produced a contraction through the release of acetylcholine from postganglionic cholinergic neurons in the duodenum of the guinea-pig in the same manner as TRH. However, the affinity of TRH-A (pD2, 4.70) toward isolated duodenum was one thousandth that of TRH (pD2, 7.74). The effects of TRH-A (10(-4)M) were abolished by 10(-7) M TRH, but only partially (about 50%) inhibited by 10(-4) M d-amphetamine. D-Amphetamine showed no stimulatory effect on the myenteric nerves. However, the duodenal response to TRH (10(-6) M) was dose dependently inhibited by d-amphetamine (10(-6), 10(-5), 10(-4) M) while the phasic and tonic contractions caused by high K+ (40 mM) or the contractile responses to acetylcholine (10(-7) M) were not blocked by d-amphetamine. These results indicate that d-amphetamine may act as an antagonist to TRH without influencing the movement of calcium ions in smooth muscle or muscarinic receptors and that contractile responses to TRH-A are mediated through TRH receptors in the myenteric cholinergic nerves.

Response to opioids of isolated small intestine in the house musk shrew, suncus murinus

1. Response to the opioids of isolated small intestine of Suncus murinus, an insectivore, was examined and compared with that of guinea-pig ileum. 2. The mechanical response to morphine and U50,488H, preferential mu- and kappa-agonist respectively, was relaxation which was antagonized by tetrodotoxin and naloxone in the Suncus small intestine. 3. Methionine5-enkephalin and D-alanine2, D-leucine5-enkephalin, both preferential delta-agonists, elicited contraction or relaxation in the Suncus small intestine, while enkephalins elicited sustained contraction in the tetrodotoxin-pretreated intestine. 4. In the guinea-pig ileum, the response to morphine and enkephalins was consistently relaxation which was antagonized by tetrodotoxin and naloxone. 5. Morphine, D-alanine2,D-leucine5-enkephalin and U50,488H inhibited electrically-evoked twitch responses of intestine in a concentration-dependent manner in both animals. 6. The relative potencies calculated from IC50 values of opioids for twitches followed the order, D-alanine2,D-leucine5-enkephalin greater than U50,488H greater than morphine in the Suncus small intestine, whereas U50,488 greater than D-alanine2,D-leucine5-enkephalin greater than morphine in the guinea-pig ileum. 7. Suncus small intestine has unique features such that D-alanine2,D-leucine5-enkephalin was most potent in inhibiting electrically-evoked twitches and that enkephalins induced contraction following tetrodotoxin pretreatment.