Rat Digestive Tract Research Articles

Stability of Two Probiotics Bacteria of Goat Milk Yoghurt in Rat Digestive Tract

Increased age will affect the bacterial population of the human digestive tract, in which many bacteria will adapt and colonize different sites. Combining probiotics and prebiotics in what has been called a synbiotic could beneficially affect the host by improving survival and implantation/adhesion of live microbial dietary supplements in the gastrointestinal flora. Probiotic bacteria are expected to survive in the digestive tract to give health effects to the host by balancing the intestinal microflora. The addition of fructo-oligosaccharides is expected to increase viability and growth of probiotics in the digestive tract. The objective of the current research is to investigate the effect of probiotic yoghurt and synbiotic yoghurt of Etawa Breed Cross Saanen (PESA) goats together with fructo-oligosaccharides, on female rats and to study the stability of probiotic bacteria in the digestive tracts. The results showed that synbiotic yoghurt intake had no significant influence (p>0.05) on ration consumption, body weight gain, feed conversion, final body weight and mortality. The synbiotic intake significantly increased the population of Bifidobacterium longum (p<0.01) and Lactobacillus acidophilus (p<0.05), but in contrast decreased (p<0.05) the population of Escherichia coli. The yoghurt synbiotic treatment over 14 days influenced probiotic bacteria’s stability as shown by the reduced population of B. longum and L. acidophilus.

Stability of Two Probiotics Bacteria of Goat Milk Yoghurt in Rat Digestive Tract

Increased age will affect the bacterial population of the human digestive tract, in which many bacteria will adapt and colonize different sites. Combining probiotics and prebiotics in what has been called a synbiotic could beneficially affect the host by improving survival and implantation/adhesion of live microbial dietary supplements in the gastrointestinal flora. Probiotic bacteria are expected to survive in the digestive tract to give health effects to the host by balancing the intestinal microflora. The addition of fructo-oligosaccharides is expected to increase viability and growth of probiotics in the digestive tract. The objective of the current research is to investigate the effect of probiotic yoghurt and synbiotic yoghurt of Etawa Breed Cross Saanen (PESA) goats together with fructo-oligosaccharides, on female rats and to study the stability of probiotic bacteria in the digestive tracts. The results showed that synbiotic yoghurt intake had no significant influence (p>0.05) on ration consumption, body weight gain, feed conversion, final body weight and mortality. The synbiotic intake significantly increased the population of Bifidobacterium longum (p

Examination of the stability of hydrophobic (CdSe)ZnS quantum dots in the digestive tract of rats

Semiconductor quantum dots show promise as alternatives to organic dyes for biological labelling because of their bright and stable photoluminescence. The typical quantum dots is CdSe because colloidal synthesis for nanocrystals of this semiconductor is well established. CdSe is usually passivated with zinc sulfide. While the cytotoxicity of bulk CdSe is well documented, questions about (CdSe)ZnS potential toxicity and behaviour in vivo remain unanswered. The distribution and stability of (CdSe)ZnS quantum dots in Wistar line rats' digestive tract were investigated. Hydrophobic quantum dots were mixed with fat or sonificated in water and administered orally. The distribution and stability of quantum dots moving through the digestive system of rats was followed by fluorescence spectroscopy. In both ways prepared quantum dots were degraded in the digestive tract of animals. Quantum dots mixed with fat were more stable and degraded more slowly than quantum dots sonificated in water. The data obtained suggest possible toxicity of (CdSe)ZnS quantum dots due to the liberation of Cd(2+).

RecombinantSaccharomyces cerevisiaeStrain Expressing a Model Cytochrome P450 in the Rat Digestive Environment: Viability and Bioconversion Activity

An innovative "biodrug" concept, based on the oral administration of living recombinant microorganisms, has recently emerged for the prevention or treatment of various diseases. An engineered Saccharomyces cerevisiae strain expressing plant P450 73A1 (cinnamate-4-hydroxylase [CA4H] activity) was used, and its survival and ability to convert trans-cinnamic acid (CIN) into p-coumaric acid (COU) were investigated in vivo. In rats, the recombinant yeast was resistant to gastric and small intestinal secretions but was more sensitive to the conditions found in the large intestine. After oral administration of yeast and CIN, the CA4H activity was shown in vivo, with COU being found throughout the rat's digestive tract and in its urine. The bioconversion reaction occurred very fast, with most of the COU being produced within the first 5 min. The gastrointestinal sac technique demonstrated that the recombinant yeast was able to convert CIN into COU (conversion rate ranging from 2 to 5%) in all the organs of the rat's digestive tract: stomach, duodenum, jejunum, ileum, cecum, and colon. These results promise new opportunities for the development of drug delivery systems based on engineered yeasts catalyzing a bioconversion reaction directly in the digestive tract.

Gene expression changes reveal patterns of aging in the rat digestive tract

Similarly to other organs, the human digestive system is adversely affected by aging presenting physiologic manifestations that include compromised absorption and secretion, decreased motility, weakened mucosal barrier and as well as a high incidence of colon cancer. As biomedical advances enable the population to live longer, our understanding of molecular events that govern aging and disease states is enhanced through methodical analyses of temporal tissue-specific gene expression profiles. Recently, DNA microarray analyses have been employed to examine age-associated transcriptional profiles in the mammalian digestive tract. Gene expression patterns revealed that the magnitude and trend of age-associated changes differ in the rat colon and duodenum. Interestingly, the expression of genes involved in energy-generating metabolic pathways was decreased in the duodenum and increased in the colon. Microarray analyses detected modulations in expression of genes associated with compromised intestinal function and propensity for colon cancer in the aged population. Furthermore, altered expression was observed for certain genes implicated in governance of aging and lifespan in other organisms suggesting intriguing commonalities across species. Thus, these studies demonstrated feasibility and usefulness of DNA microarrays for identifying pathways involved in the molecular pathophysiology of the aging process and lifespan control in complex organisms.

Effectiveness of Bombesin and Saccharomyces boulardii Against the Translocation of Candida albicans in the Digestive Tract in Immunosuppressed Rats

We investigated the effects of bombesin on disseminated candidiasis, and compared the effectiveness of bombesin with Saccharomyces boulardii against Candida albicans translocation from the gastrointestinal tract in immunosuppressed rats. Sixty rats were divided into five groups of 12. Group 1 was given only a laboratory pellet diet and water during the experiments; the other four groups were orally inoculated with C. albicans; and groups 3, 4, and 5 were also given prednisolone intraperitoneally. The treatment groups consisted of group 4, given S. boulardii orally, and group 5, given bombesin subcutaneously. The rats were killed after 10 days, and the large bowel, liver, spleen, and kidneys were removed for microbiological and histopathological examination. Blood samples were taken to measure tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta) levels, and the results were compared. The number of translocated C. albicans colonies from the gastrointestinal tract and the serum TNF-alpha and IL-beta levels were significantly lower in groups 4 and 5 than in group 3 (P < 0.05). Histological analysis revealed that the bombesin-treated group (group 5) had significantly less mucosal ulceration and submucosal inflammation in the large bowel, less inflammation and necrosis in the liver, and less inflammation of the Bowman capsules in the kidney than the S. boulardii-treated group (group 4) (P < 0.05). These findings show that both S. boulardii and bombesin inhibit the translocation of C. albicans from the gastrointestinal tract, although mucosal ulceration, submucosal inflammation in the large bowel, and dissemination in the liver and kidneys were significantly less severe in the bombesin-treated immunosuppressed rats.

Expression of LAT1 and LAT2 amino acid transporters in human and rat intestinal epithelial cells

The present study evaluated the presence of LAT1 and LAT2 amino acid transporters in human Caco-2 cells and rat IEC-6 cells along the mucosa of the rat digestive tract. The LAT1 cDNA was amplified by PCR using two sets of primers (one specific for rat LAT1 and another simultaneously specific for human, rat and mice). The LAT2 cDNA was amplified by PCR using one set of primers simultaneously specific for human, rat and mice LAT2. The presence of LAT1 and LAT2 protein was examined by means of immunoblotting using an antibody raised against the rat LAT1 and mouse LAT2. Caco-2 and IEC-6 cells, as well as the rat intestinal mucosa, are endowed with both LAT1 and LAT2 transporter transcripts and protein. LAT1 protein is most abundant in IEC-6 cells, which is in agreement with functional data previously reported. The findings in the rat intestinal mucosa indicate that LAT1 protein is most abundant in the colon and its abundance markedly decreases at the level of jejunum and ileum, which contrast with relative homogeneous presence of LAT2 across the digestive tract. In conclusion, Caco-2 and IEC-6 cells, as well as the rat intestinal mucosa, are endowed with both LAT1 and LAT2 amino acid transporter transcripts and protein.

Models of phage growth and their applicability to phage therapy

Phage therapy is complicated by the self-replicating nature of phage. It is difficult to extrapolate from in vitro phage growth data to in vivo expectations, difficult to interpret in vivo data and difficult to generalize from one in vivo situation to another. Various generic models of phage growth have been used as the theoretical basis for understanding the kinetics of phage therapy. Here, we have experimentally tested the efficacy of such simple models to predict, qualitatively and quantitatively, the growth of phage and the phage proliferation threshold in vitro. Naturally occurring, antibiotic-resistant bacteria were used to measure the growth of phage in vivo. In homogenous, in vitro environments, the models were predictive of T4 phage growth on Escherichia coli RR1. However, the models were not able to predict growth of T4 phage or K1-5 phage in the more complex environment of the rat's digestive tract. To explore fully the kinetics of phage therapy, more complex models need to be devised. We suggest that it may be necessary to consider and model the interactions between phage growth parameters and bacterial growth parameters.

P2X2 and P2X3 purinoceptors in the rat enteric nervous system.

Adenosine 5'-triphosphate receptors are known to be involved in fast excitatory postsynaptic currents in myenteric neurons of the digestive tract. In the present study, the distribution of P2X2 and P2X3 receptor mRNA was examined by in situ hybridisation while P2X2 and P2X3 receptor protein was localised by immunohistochemical methods. In addition, P2X2 and P2X3 receptors were colocalised with calbindin and calretinin in the myenteric and submucosal plexus. P2X2- and P2X3- immunoreactive neurons were found in the myenteric and submucosal plexuses throughout the entire length of the rat digestive tract from the stomach to the colon. Approximately 60%, 70% and 50% of the ganglion cells in the myenteric plexus of the gastric corpus, ileum and distal colon, and 56% and 45% in the submucosal plexus of the ileum and distal colon, respectively, showed positive immunoreactivity to the P2X2 receptor. Approximately 10%, 2% and 15% of the ganglion cells in the myenteric plexus of the gastric corpus, ileum and distal colon, and 62% and 40% in the submucosal plexus of the ileum and distal colon, respectively, showed positive immunoreactivity to the P2X3 receptor. Double-labelling studies showed that about 10-25% of the neurons with P2X2 immunoreactivity in myenteric plexus and 30-50% in the submucosal plexus were found to express calbindin or calretinin. About 80% of the neurons with P2X3 receptor immunoreactivity in the myenteric plexus and about 40% in the submucosal plexus expressed calretinin. Approximately 30-75% of the neurons with P2X3 receptor immunoreactivity in the submucosal plexus expressed calbindin, while none of them were found to express calbindin in the myenteric plexus.

Digestion of Maize Sphingolipids in Rats and Uptake of Sphingadienine by Caco-2 Cells

We investigated the digestion of cerebrosides of plant origin prepared from maize, focusing especially on the digestive fates of trans-4, cis-8- and trans-4, trans-8-sphingadienine, which are common in higher plants. In the small intestinal mucosa and cecal contents of rats, the cerebrosidase activity at pH 5.2 toward the glucosyl linkage in maize cerebrosides (glucosylceramides) was similar to that in cerebrosides of mammalian origin. Similarly, the ceramidase activity toward the amide linkage in ceramides prepared from maize cerebrosides at pH 7.0 was the same as that toward ceramides of mammalian origin. In addition, maize cerebrosides were hydrolyzed to ceramide and free sphingoid bases in the digestive tract of rats after oral administration. To further evaluate the uptake by enterocytes of 4,8-sphingadienine, we used differentiated Caco-2 cells, derived from human colonic carcinoma, as a model of intestinal epithelial cells. The accumulation of sphingoid bases in Caco-2 cells incubated with each isomer of sphingadienine was lower than that after incubation with sphingosine (P < 0.05). Verapamil, a P-glycoprotein inhibitor, increased the accumulation of each sphingadienine but not of sphingosine, suggesting that the efflux of sphingadienine of plant origin, but not sphingosine of mammalian origin, was affected by P-glycoprotein. The digestibility of maize cerebrosides appears similar to that of cerebrosides of mammalian origin, but the metabolic fate of sphingoid bases of plant origin within enterocytes differs from that of sphingosine. Isomers of 4,8-sphingadienine degraded from dietary plant cerebrosides appear to be poorly absorbed from the digestive tract.

Localization and role of heme oxygenase in digestive tract of rat

Localization and role of heme oxygenase in digestive tract of rat

Colocalization of NO and VIP in neurons of the submucous plexus in the rat intestine

Since very few previous studies have carried out the quantitative analysis for the colocalization of nitric oxide (NO) and vasoactive intestinal peptide (VIP) in the submucous neurons in the rat digestive tract, we applied in vivo treatment of colchicine to enhance the immunoreactivity and examined the colocalization of NO synthase (nNOS) and VIP in neurons of the submucous plexus throughout the rat digestive tract. The density of nNOS-containing neurons in the submucous plexus in the stomach corpus (103±25 cells/cm 2, n=3) and that in the antrum (157±9 cells/cm 2, n=3) were significantly lower than those in small and large intestine. However no difference was detected in the cell density among duodenum (1967±188 cells/cm 2, n=3), jejunum (2640±140 cells/cm 2, n=3), ileum (2070±42 cells/cm 2, n=3), proximal colon (2243±138 cells/cm 2, n=3) and distal colon (2633±376 cells/cm 2, n=3). The proportion of nNOS-immunoreactive (IR), nNOS/VIP-IR and VIP-IR neurons to the total number of submucous neurons was examined. nNOS/VIP-IR neurons comprised 45–55% of total number of submucous neurons from the duodenum to the proximal colon, however those comprised 66.4±5.1% in the distal colon. The results showed that the dense distribution of nNOS-containing neurons was found in the submucous plexus throughout the small and large intestine, and large population of submucous neurons co-stored nNOS and VIP.

Gluconoacetobacter hansenii에 의해 생산된 섬유소 섭취가 흰쥐의 소화기관과 지질대사에 미치는 영향

본 실험은 초산균인 Gluconoacetobacter hansenii에 의해 코코넛 배지에서 생성된 막인 bacterial fiber가 흰쥐의 장 전반과 소장의 이당류 효소활성 및 체내 지질상태에 미치는 영향을 조사하였다. 이를 위하여 흰쥐를 0.5% 콜레스테롤 식이로 사육하였으며 섬유소원으로 bacterial fiber를 식이의 2%로 첨가하였으며, 효과를 비교할 실험군들에게는 섬유소원으로 식물성 cellulose와 pectin을 같은 수준으로 식이에 첨가하여 사육하였다. 실험 4주간 동안 식이섭취량 및 체중 증가량은 세 군간에 차이가 없었다. 소장의 길이, 무게 등은 군간에 유의적인 차이는 없었으나 소장점막 sucrase활성이 bacterial fiber군에서 감소하였고, maltase 활성도 감소하는 경향이었다. 맹장의 무게가 bacterial fiber군에서 증가하였으나, 결장의 무게에는 영향이 없었고, 결장 내용물 및 건조분변량이 감소하였다. 혈장 총 콜레스테롤 농도는 세 군 중에서 pectin군이 유의적으로 낮았고 HDL-콜레스테롤 함량은 반대로 pectin군에서 높았다. 혈장과 간조직의 중성지방 함량은 모두 bacterial fiber군에서 다른 군들에 비하여 유의적으로 낮았다. 간조직 콜레스테롤 함량은 pectin군에서 낮았다. 간조직의 상대적 무게는 군간에 차이가 없었고, 혈장 GOT 및 GPT 활성도 bacterial fiber군에서 다른 두 군에 비하여 차이가 없었다. 본 연구의 결과, 실험에서 사용한 bacterial fiber는 체내의 중성지방 수준을 낮추어 지질 상태를 개선 할 수 있는 가능성을 보여 주어 이에 대한 기전과의 작용의 활용에 대한 연구가 요망된다. 본 실험에서는 콜레스테롤 및 소장의 구조와 소화효소에는 큰 영향을 주지 않았으나 사용하는 섬유소의 수준을 보다 높이거나 다른 섬유소와 복합적으로 사용하는 경우, 또 동물의 나이에 따라 다른 결과가 도출될 수도 있으리라고 사료되어 앞으로의 지속적인 연구가 필요하다.

Molecular analysis of streptogramin resistance in enterococci

AbstractThe new semi-synthetic streptogramin antibiotic combination quinupristin/dalfopristin (Synercid®) is a promising alternative for a treatment of infections with multiple resistant gram-positive pathogens, e.g. glycopeptide- and multi-resistant Enterococcus faecium. Streptogramins consist of two unrelated compounds, a streptogramin A and B, which act synergistically when given in combination. Mechanisms conferring resistance against both components are essential for resistance against the combination in E. faecium. In this species resistance to streptogramin A compounds is mediated via related acetyltransferases VatD and VatE. Resistance against streptogramins B is either encoded by the widespread ermB gene cluster conferring resistance to macrolide-lincosamide-streptogramin B antibiotics or via expression of the vgbA gene, which encodes a staphylococcal-type lactonase. E. faecalis is intrinsically resistant to streptogramins.Due to a wide use of streptogramins (virginiamycins S/M) in commercial animal farming a reservoir of streptogramin-resistant E. faecium isolates had already been selected. Determinants for streptogramin resistance are localized on plasmids that can be transferred into an E. faecium recipient both in vitro in filter-matings and in vivo in the digestive tracts of rats. Hybridization and sequencing experiments revealed a linkage of resistance determinants for streptogramins A and B on definite plasmid fragments.

Expression patterns of 5-HT 7 receptor isoforms in the rat digestive tract

In this study, we determined the expression patterns of the 5-HT 7 receptor isoforms at the mRNA level in the rat jejunum and other regions of the rat digestive tract using reverse transcriptase-PCR. In control studies using the rat thalamus, we detected all three known isoforms of the rat 5-HT 7 receptor, 5-HT 7(a), 5-HT 7(b) and 5-HT 7(c). In addition, we also detected in the thalamus a new PCR product that we provisionally name 5-HT 7(e). All four isoforms of the 5-HT 7 receptor were present throughout the rat digestive tract. However, the spatial distribution and levels of expression of the different isoforms varied between regions of the rat digestive tract. All three known isoforms and the provisional isoform were found in the jejunum, ileum, stomach fundus, oesophagus and colon. The level of expression of the isoforms varied with all four being most consistently found in the ileum. The 5-HT 7(a) receptor isoform was predominantly expressed in the rat digestive tissue and 5-HT 7(b), 5-HT 7(c) and 5-HT 7(e) were only sometimes expressed but always accompanying the receptor isoform 5-HT 7(a).

Immunohistochemical and in situ hybridization studies of gonadotropin releasing hormone (GnRH) and its receptor in rat digestive tract

GnRH(LH-RH) is first discovered in the hypothalamus and found to have a role in regulation of reproduction. With the study on it deepening, GnRH was demonnstrated that it also exists in a number of organs beyond the hypothalamus and acts on extrapituitary organs. To study whether digestive tract synthesizes GnRH and its receptor and, if it does, by what cells. In the experiment, the locallizations of GnRH and its receptors in rat digestive tract were studied using immunohistochemistry and in situ hybridization. The parietal cells of gastric gland, the villous and glandular epithelium in small and large intestine and parasympathetic ganglion cells of myenteric plexus showed GnRH immunoreactivity; GnRH mRNA hybridization signal was detected. The epithelium of gastric pit and the cells above in digestive tract showed GnRH receptor immunoreactivity; GnRH receptor mRNA hybridization signal was detected. The immunoreactive and signal materials distributed in cytoplasm of all positive cells, with nuclei being immunonegative and with no hybridization signal. These results suggested that the digestive tract can produce GnRH and express GnRH receptor; GnRH may also be a gastrointestinal hormone.

Possible role of K-ras oncogene in mutagenic activity of ethylnitrosourea on lymphocyte hyperproliferation in rat colon.

N-ethyl-N-nitrosourea (ENU) is a potential carcinogenic agent that is commonly used in industry. Therefore the present study aimed to find out the possible effects of this agent on the rat digestive tract especially on the colon. We have studied the complementary mutation activity of the exon 2 of K-ras oncogene by ENU treatment in rat colonal tissue. While the two experimental group rats were injected once a week with 20 mg/kg and 300 mg/kg body weight-body weight with ENU (i.p.), the last experimental group was administered only with PEG and the control group animals received no treatment. Following 45 weeks, all animals were sacrificed and colonal tissues were obtained. Tissues were processed for light and electron microscopy and also for molecular biological analyses. While no colonal tumour development was observed in the control and in the PEG treated group, an extensive tumour development was seen in a wide range of tissues in the high dose ENU treated group. The light and electron microscopical examination of the rat colonal tissue revealed a lymphocyte hyperproliferation in the submucosal region, an increased number of polymorphonuclear leukocytes (PMLs) and occasional epithelial lesions. The mutation analyses of exon 2 of K-ras by HpaII demonstrated more than one recognition sites in the ENU treated group whereas there was only one enzyme cognate site in the control group.

Effect of raw and heat-moisture-treated high-amylose corn starches on the process of digestion in the rat digestive tract

High-amylose corn starch (HAS) is widely known as a resistant starch foodstuff. We developed heat-moisture-treated high-amylose corn starch (HMT-HAS) that was more resistant to enzymatic hydrolysis. Resistant starch contents of HAS and HMT-HAS using the enzymatic-gravimetric method were found to be 30% and 65% respectively. Rats were given 10% ordinary corn starch (CS), HAS or HMT-HAS by meal feeding for 10 days. The caecum contents increased and the caecal pH was lower after their diets were supplemented with HAS and HMT-HAS. Starch contents increased in the upper and the lower small intestine with HAS and HMT-HAS. Caecal starch with HAS and HMT-HAS was more than that with CS. Particularly, caecal starch with HMT-HAS was seven times more than that with HAS. There were no differences in starch content in the large bowel between CS and HAS, but the content increased with HMT-HAS. These results suggested that HAS and HMT-HAS were resistant to digestion and absorption in the small intestine, and any indigestible starches reached the caecum. In the caecum, HAS was hydrolysed almost completely by intestinal bacteria; however, some HMT-HAS escaped bacterial hydrolysis. This escaped HMT-HAS reached the large bowel and was excreted in the faeces.

Expression profile of saccharide epitope CaMBr1 in normal and neoplastic tissue from dogs, cats, and rats: implication for the development of human-derived cancer vaccines.

CaMBr1 is a blood group-related tumour-associated antigen, whose pattern of expression provides a therapeutic window for passive or active immunotherapy and points to the promise of a vaccine against carcinomas overexpressing this antigen. In this context, an animal model that closely mimics the human situation would be extremely useful. We, therefore, utilised the murine monoclonal antibody MBr1, which defines CaMBr1, as a useful probe to detect the molecule targeted for vaccine development on canine and feline spontaneous breast and uterus tumours and on their normal counterparts, and on rat normal tissues and carcinoma cell lines. Immunoperoxidase staining of cryostat sections revealed homogeneous CaMBr1 expression only in normal feline uterus and a uterus papilloma, whereas MBr1 reactivity was very weak and heterogeneous in normal (1/3 and 1/3) and tumour (1/10 and 1/6) breast tissues from dogs and cats, respectively. In contrast, the data obtained in rat tissues were reproducible in the strains tested and showed that CaMBr1 was expressed in all epithelial tissues of the digestive tract, although with variable intensities. Monoclonal antibody staining appeared to correspond to membrane-bound structures as well as mucinous secretions. Similarly, secretion products of lactating mammary glands expressed CaMBr1. The spectrum of expression on rat digestive tract was broader than that in humans but the specificity of MBr1 reactivity was confirmed by competition assay with a synthetic tetrasaccharide that mimics the CaMBr1 antigen. On FACS analysis, only one of two clonal derivatives of the rat breast carcinoma line RAMA 25 expressed CaMBr1, and a negative cell subset was evident in repeated experiments. By contrast, both colon carcinoma lines, DHD/K12 and CC531, showed staining with MBr1, albeit at different levels of intensity, and no evidence of a negative subset. The cell line CC531 maintained or even increased CaMBr1 expression levels following transplantation in syngeneic immunocompetent animals. Our data suggest the usefulness of the rat as a test model for vaccines against human cancers overexpressing the CaMBr1 antigen.

PACAP in visceral afferent nerves supplying the rat digestive and urinary tracts.

PACAP in visceral afferent nerves supplying the rat digestive and urinary tracts.