Localisation and expression of TRPV6 in all intestinal segments and kidney of laying hens

1. The aim of this study was to investigate the localisation and expression of the epithelial Ca2+ channel TRPV6 (transient receptor potential vanilloid channel type 6) in different intestinal segments and kidney of laying hens during peak lay. 2. Immunohistochemical analysis of the intestine indicated that TRPV6 was localised to the brush-border membranes of the duodenum, jejunum, ileum, caecum, and rectum. Expression was weaker in the rectum, and little or no expression was found in crypt and goblet cells. In addition, TRPV6 mRNA was quantified amongst different intestinal segments, and expression was highest in the duodenum and jejunum. Furthermore, Western blotting indicated that the duodenum expressed the greatest amount of TRPV6 and the rectum the least with the other segments expressing intermediate levels. 3. In the kidney, distinct immunopositive staining for TRPV6 was detected at the apical domain of the distal convoluted tubules (DCT) and medullary connecting tubules (CNT). Interestingly, distribution of TRPV6 extended to the proximal convoluted tubules (PCT). Furthermore, the kidney expressed lower TRPV6 mRNA and protein levels compared with that in the duodenum. 4. In conclusion, the epithelial Ca2+ channel TRPV6 is strongly expressed in the apical cells of the entire intestine and the renal tubules, suggesting that active Ca2+ transcellular transport plays a crucial role in dietary calcium (re)absorption in laying hens.     

鸡肠道SGLT1和GLUT2 mRNA表达的组织特异性研究

运用相对定量RT-PCR方法，研究不同肠段Arbor Acre（AA）肉鸡肠道葡萄糖吸收转运主要载体SGLT1和GLUT2mRNA表达的组织特异性。结果发现。随着肠道空间位置的后移，SGLT1 mRNA的表达量逐步降低。十二指肠SGLT1 mRNA的丰度比结直肠高76．19Vo，差异极显著（P〈0．01）；而空肠和回肠SGLT1 mRNA的表达量分别比结直肠高42．86％和38．10％，差异不显著（P〉0．05），但有提高的趋势（P值分别为0．06和0．07）。十二指肠与空肠和回肠相比，SGLTlmRNA的表达量虽然分别高23．33％和27．59％，但差异不显著（P值分别为0．18和0．10）。相对定量分析表明，十二指肠和空肠GLUT2 mRNA丰度非常接近，差异不显著（P〉0．05）。定性研究显示，十二指肠与空肠GLUT2 mRNA丰度高于回肠和结直肠。鸡肠道SGLT1和GLUT2 mRNA表达的组织特异性之生理功能，有待于进一步研究。     

Penetration in vitro of newly excysted juvenile flukes of Japanese Fasciola sp. through ligated intestines of rabbits, mice, rats and chickens.

Ligated intestines of rabbits, mice, rats and chickens were used to examine the penetration of newly excysted juvenile flukes of Japanese Fasciola sp. in vitro. In rabbit intestines, the penetration rate was relatively high in the rectum and duodenum. Penetration rates in the jejunum, ileum, cecum and colon were comparable to those in the rectum and duodenum, although it was lower in the appendix. In the case of mouse, juvenile flukes penetrated the duodenum, jejunum, cecum, and rectum at considerably high rates. In rat intestine, penetration by flukes was less in the duodenum and rectum, although flukes were detected in the jejunum. In chicken intestine, flukes barely penetrated the duodenum, jejunum and rectum. Consequently, newly excysted flukes of Fasciola sp. seem to penetrate any region of the intestine in rabbits and mice. In rats, the middle small intestine may be the site suitable for flukes to penetrate. In chickens, the difficulty in penetration of the intestinal wall may be one of the reasons why chickens are scarcely infected with Fasciola sp.     

Expression and localization of BCRP,MRP1 and MRP2 in intestines,liver and kidney in horse

Tydén, E., Bj?rnstr?m, H., Tjälve, H., Larsson, P. Expression and localization of BCRP, MRP1 and MRP2 in intestines, liver and kidney in horse. J. vet. Pharmacol. Therap. doi: 10.1111/j.1365‐2885.2009.01140.x. The gene and protein expression and the cellular localization of the ABC transport proteins breast cancer resistance protein (BCRP), multidrug resistance‐associated protein 1 (MRP1) and multidrug resistance‐associated protein 2 (MRP2) have been examined in the intestines, liver and kidney in horse. High gene and protein expression of BCRP and MRP2 were found in the small intestines, with cellular localization in the apical membranes of the enterocytes. In the liver, MRP2 was present in the bile canalicular membranes of the hepatocytes, whereas BCRP was localized in the cytoplasm of hepatocytes in the peripheral parts of the liver lobuli. In the kidney both BCRP and MRP2 were predominantly present in the distal tubuli and in the loops of Henle. In most tissues, the gene and protein expression of MRP1 were much lower than for BCRP and MRP2. Immunostaining of MRP1 was detectable only in the intestines and with localization in the cytoplasm of enterocytes in the caecum and colon and in the cells of serous acini of Brunner’s glands in the duodenum and the upper jejunum. The latter cells were also stained for BCRP, but not for MRP2. Many drugs used in horse are substrates for one or more of the ABC transport proteins. These transporters may therefore have important functions for oral bioavailability, distribution and excretion of substrate compounds in horse.     

GHSR—1a在奶山羊胃肠道的分布与表达

试验采用免疫组织化学、Real—timePCR和Western blotting方法测定ghrelin的功能性受体GHSR-1a（Growth hormone seeretagogue receptor-1a,GHSR-1a）在奶山羊胃肠道的分布和表达。免疫组织化学结果显示,GHSR—1a免疫阳性细胞广泛分布于奶山羊胃肠道。在皱胃主要定位于黏膜层和肌层;瘤胃、网胃和瓣胃黏膜层及肌层中也可见GHSR-1a免疫阳性细胞;在小肠主要位于十二指肠、空肠和回肠的黏膜层、黏膜下层和肌层;在结肠、盲肠和直肠GHSR—1a免疫阳性细胞也有广泛分布;GHSR—1a主要表达于内在神经丛神经细胞、胃底腺上皮细胞、肠腺上皮细胞、复层鳞状上皮细胞、平滑肌细胞中。real—timePCR和Westernblotting结果显示,皱胃、十二指肠、空肠、回肠、盲肠、结肠和直肠GHSR—1a的表达水平相对较高,显著高于瘤胃、网胃和瓣胃的表达（P〈0．05）。结果表明,ghrelin可能通过GHsR-1a对奶山羊胃肠功能具有重要的调节作用。     

Expression and localisation of calbindin D28k in all intestinal segments of the laying hen

1. The aim of the present study was to investigate expression and localisation of a 28-kDa calcium-binding protein (CaBP-D28k) related to active calcium (Ca) absorption, in the entire intestine of egg-laying hens. 2. Western blotting analysis showed that the entire intestine expressed CaBP-D28k to the following degree: duodenum > jejunum > caecum > ileum > colon. Immunohistochemistry showed strong CaBP-D28k localisation in enterocytes along the villus tip-crypt axis in the duodenum and in villus tips in the caecum and colon. The jejunum and ileum had moderate localisation with respect to the number of immunoreactive cells and staining intensity. 3. These results suggest that laying hens actively absorb Ca in both the large and small intestines.     

Expression of CD147 and monocarboxylate transporters MCT1, MCT2 and MCT4 in porcine small intestine and colon

Lactate, formed mainly in the stomach and small intestines, and short-chain fatty acids (SCFAs) formed in the colon, are ionised and require transporter proteins such as monocarboxylate transporters (MCTs) for absorption. The amounts of MCT1, MCT2, MCT4 and CD147, an ancillary protein for MCT1 and MCT4, were measured by immunoblotting the small intestine and colon of 40 pigs (Landrace, Yorkshire and LandracexYorkshire). MCT1 and MCT4 were found in both small intestine and colon, but MCT2 only in the small intestine. In both small intestine and colon, Yorkshire pigs had more CD147 than Landrace pigs, while no interbreed differences were found in MCT isoforms. Since CD147 is essential for the activity of MCT1 and MCT4, the breed difference suggests that MCT activity is higher in Yorkshire than in Landrace pigs. The absence of MCT2 in the colon suggests that it is mainly a lactate transporter, while MCT1 and MCT4 facilitate the transport of both lactate and SCFA.     

肉鸡肠道NHE2 mRNA表达的组织特异性与发育性变化

选用遗传背景相同的1日龄父母代雄性Arbor Acre（AA）肉雏鸡120羽,随机分为4个重复,采用相对定量RT-PCR方法,以30日龄AA肉鸡肠道RNA为模板,研究肉鸡肠道钠/氢交换载体2（Sodium hydrogen exchanger 2,NHE2）mRNA表达的组织特异性;以AA肉鸡十二指肠和空肠RNA为模板,研究肉鸡肠道NHE2mRNA表达的发育性变化。结果显示：①AA肉鸡十二指肠和空肠NHE2 mRNA的表达丰度显著高于回肠和结直肠（P〈0.05）,而十二指肠和空肠之间、回肠和结直肠之间无显著差异（P〉0.05）;②AA肉鸡NHE2 mRNA在十二指肠及空肠中的表达具有相同的发育模式,2～16日龄升高,30～44日龄下降,55日龄略微回升;在16和30日龄时的表达丰度显著高于2、44和58日龄（P〈0.05）。以上结果说明：AA肉鸡肠道近端NHE2 mRNA的表达丰度显著高于远端（P〈0.05）。AA肉鸡十二指肠及空肠NHE2 mRNA的表达具有相同的发育模式,表明NHE2mRNA表达受到发育阶段的调控,且在十二指肠和空肠间具有稳定性。     

肉鸡肠道PepT1 mRNA表达的肠段差异性与发育性变化

选用遗传背景相同的1日龄父母代雄性Arbor Acre(AA)鸡和父母代雄性岭南黄肉雏鸡各120羽,采用相对定量RT-PCR方法,以30 d时岭南黄鸡肠道样品为模板,研究肉鸡肠道寡肽转运载体1(Peptide transporter 1,PepT1)mRNA表达的肠段差异性;以AA肉鸡和岭南黄鸡十二指肠和空肠样品为模板,研究不同品种肉鸡肠道PepT1 mRNA表达的发育性变化。结果显示:(1)岭南黄肉鸡肠道PepT1 mRNA的表达丰度从十二指肠、空肠、回肠到结直肠依次降低,其中十二指肠显著高于结直肠(P<0.05);(2)AA鸡和岭南黄肉鸡PepT1 mRNA在十二指肠及空肠中的表达具有相同的发育模式,16 d表达丰度最高,16~44 d逐渐下降,58 d略微回升;不同基因型之间PepT1 mRNA丰度比较,AA鸡和岭南黄肉鸡两品种间PepT1 mRNA的表达没有显著差异(P>0.05)。以上结果说明:(1)随着肠道空间位置的后移,岭南黄肉鸡肠道PepT1 mRNA的表达丰度逐渐降低,其中十二指肠的表达丰度显著高于结直肠(P<0.05);(2)不同品种肉鸡十二指肠及空肠PepT1 mRNA的表达具有相同的发育模式,且同日龄两品种间的表达丰度未见明显差异,表明PepT1 mRNA表达受到发育阶段的调控,但品种间具有稳定性。     

Immunohistochemical study of endocrine cells in the gastrointestinal tract of the barking deer, Muntiacus muntjak

The morphology, distribution and relative frequencies of endocrine cells in the gastrointestinal tract of the barking deer (Muntiacus muntjak) were investigated. The immunohistochemical method employed used seven types of antisera against chromogranin, serotonin, gastrin, cholecystokinin, somatostatin, glucagon and insulin. Chromogranin and serotonin immunoreactive (IR) cells were found throughout the gastrointestinal tract. Conversely, gastrin-IR cells were only found in the oxyntic gland, pyloric gland and duodenum, while cholecystokinin-IR and somatostatin-IR cells were detected in the oxyntic gland, pyloric gland and small intestines. Somatostatin-IR cells were also seen in the caecum. Glucagon-IR cells were found in all parts of the gastrointestinal tract apart from the colon and rectum. No insulin-IR cells were found in the gastrointestinal tract of this species. The cells in the small intestine were generally spindle shaped with long cytoplasmic processes ending in the lumen (open type), while in the stomach and large intestine, they were occasionally round or spherical in shape (closed type). An uncommon distribution pattern of endocrine cells in the gastrointestinal tract of the barking deer was noted for cholecystokinin- and glucagon-IR cells.     

Fatty acid‐binding protein expression in the gastrointestinal tract of calves and cows

Fatty acid‐binding protein (FABP) has high affinity for long‐chain fatty acids and appears to participate in the metabolism and intracellular transport of lipids. Liver‐ and intestinal‐type FABP (L‐FABP and I‐FABP, respectively) are expressed in the small intestine. However, in the gastrointestinal tract of ruminants, expression and localization of FABPs are unknown. In this study, we investigated the expression of I‐FABP and L‐FABP in the gastrointestinal tract of cattle. I‐ and L‐FABP had higher messenger RNA (mRNA) and protein expression levels in the duodenum and jejunum relatively to other gastrointestinal regions in both calves and cows. Furthermore, L‐FABP mRNA and protein expression were high in the colon. Both these protein types were confirmed to be in the cytosol of jejunal epithelial cells, where they were found in the villi rather than in the crypts. We concluded that duodenal and jejunal FABPs might be involved in the metabolism of fatty acids mainly in epithelial cells in cattle.     

The presence of EGF- and IGF-1-receptors in the small intestine of fetal, neonatal and weaned piglets

During development, gut morphology and functions change. Some of these are mediated by milk-borne factors, e.g. insulin-like growth factor-1 (IGF-1) and epidermal growth factor (EGF), of which the effects are in part dictated by the distribution of their receptors (R).

EGFR- and IGF-1R-immunoreactivity (IR) was evaluated in samples of the fetal, neonatal and weaned porcine small intestine.

EGFR-IR first appeared in the neonatal duodenum and was located basolaterally in villar and to a lesser extent in cryptal enterocytes. In weaned pigs, EGFR appeared apically in enterocytes with a region-dependent distribution along the villus–crypt axis. IGF-1R-IR was observed in smooth muscle cells in each age group. From the last trimester of gestation onward IGF-1R-IR was seen at the apical and basolateral side of villar enterocytes and in submucosal arterioles.

Thus, the age- and region-dependent presence of EGFR and IGF-1R has to be taken into account when evaluating effects of EGF and IGF-1 on growth or repair.     

Differential Expression of Monocarboxylate Transporter 1 and Ancillary Protein CD147 in Red Blood Cells of Show Jumping Horses

We compare the expression levels of the lactate transporter complex consisting of the lactate transporter, monocarboxylate transporter 1 (MCT1), and its ancillary protein, cluster of differentiation 147 (CD147), in the membranes of red blood cells (RBCs) from two breeds of jumping horses and associate the expression levels of these proteins with their jumping ability. The expression levels of MCT1 and CD147 proteins on the membranes of RBCs collected from 30 show jumping horses of two different breeds were quantified: the Brazilian Sport Horses (n = 17) and the European Warmbloods (n = 13). The levels of MCT1 and CD147 in the RBC membranes were measured by western blot using horse-specific antibodies. Statistical analyses included unpaired Student t test and chi-squared test. According to the expression levels of MCT1 and CD147 proteins, 88% of the Brazilian Sport Horses were categorized as high lactate transporters (HTs) and the remaining 12% as low lactate transporters (LTs). The opposite was found for the European Warmbloods, where most animals (77%) were classified as LTs and the remaining animals (23%) were classified as HTs. Brazilian Sport Horses express statistically significantly higher levels of CD147 and MCT1 than European Warmbloods. The classification of horses considering the expression of proteins involved in the ability to transport lactate through the complex MCT1-CD147 seems to be breed dependent, with horses that are able to jump higher obstacles showing lower expression of the MCT1-CD147 complex in their RBCs.     

猪肠道碱性氨基酸转运载体(CAT1)mRNA表达的组织特异性和发育性变化

本研究旨在探讨不同品种猪肠道碱性氨基酸转运载体（CAT1）mRNA表达的组织特异性和发育规律。试验选取1、7、26、30、60、90和150日龄长白和蓝塘公猪各5头（同一品种同一日龄且体重接近）,共70头,测体重后屠宰,采集十二指肠、空肠、回肠和结肠组织样品。以18S基因为内标,用实时荧光定量RT-PCR法（SYBR Green Ⅰ试剂盒）检测CAT mRNA在60日龄长白猪十二指肠、空肠、回肠和结肠表达的组织特异性,以及在长白和蓝塘猪不同日龄其十二指肠、空肠、回肠表达的发育性变化。结果显示：60日龄长白猪CAT1 mRNA的表达丰度从十二指肠到回肠呈逐渐升高的趋势,到结肠开始下降,回肠极显著高于其他3个肠段（P〈0．01）,十二指肠最低;十二指肠和空肠CAT mRNA的表达在1～26d（即哺乳期）都呈上升的趋势,随后都开始有所下降;蓝塘猪十二指肠CAT1 mRNA的表达丰度在150d时显著低于其他各个阶段（P〈0．05）,而长白猪90和150d都显著低于其他各阶段（P〈0．05）;空肠CAT1 mRNA的表达在26～150d各阶段都差异不显著,而26d显著高于1和7d（P〈0．05）;1～60d长白和蓝塘猪回肠CAT1 mRNA的表达都呈逐渐上升的趋势,60d后都显著下降（P〈0．05）。两品种猪不同日龄时十二指肠和空肠CAT1 mRNA的表达量都没有显著差异（P〉0．05）;长白猪回肠CAT mRNA表达在26d时显著高于蓝塘猪（P〈0．05）;在90和150d时,长白猪都显著低于蓝塘猪（P〈0．05）,其他各阶段没有显著差异。结果说明,CAT mRNA在不同肠段及不同发育阶段的表达存在明显的差异,这可能与肠腔中氨基酸的浓度和氨基酸的需要水平及相关激素水平有关。     

牛瘤胃上皮细胞中挥发性脂肪酸吸收相关蛋白在白细胞介素10诱导下的差异表达研究

本研究旨在探讨炎症因子（白细胞介素10, IL-10）对牛瘤胃上皮细胞中挥发性脂肪酸（VFA）吸收相关蛋白基因表达的影响。试验采用随机区组设计,在瘤胃上皮细胞体外培养条件下,研究50 ng·mL^-1浓度的IL-10对牛瘤胃上皮细胞中核因子（NF-κB）、假定阴离子转运蛋白1（PAT1）、阴离子交换蛋白2（AE2）、单羧酸转运蛋白1（MCT1）、单羧酸转运蛋白4（MCT4）、钠氢离子交换蛋白1（NHE1）基因表达的影响。结果表明,在中性条件下,IL-10添加显著下调了NF-κB、PAT1、MCT1、NHE1基因的表达（P<0.05）,AE2基因的表达显著性上调（P<0.05）,而对MCT4基因的表达没有显著影响。在酸性条件下,IL-10添加组的NF-κB基因表达被显著性抑制（P<0.05）,而PAT1、MCT1、MCT4基因表达显著上调（P<0.05）。pH5.5组与pH7.2组比较,pH5.5组NF-κB基因表达显著性上调（P<0.05）。结果提示,在炎症条件下,白细胞介素10作为一种抗炎因子,可以缓解瘤胃上皮细胞炎症反应从而促进VFA吸收相关蛋白基因的表达。     

Differences in monocarboxylic acid transporter type 1 expression in rumen epithelium of newborn calves due to age and milk or milk replacer feeding

The aim of the present study was to investigate whether besides age and solid feed intake, monocarboxylic acid transporter type 1 (MCT1) expression in the rumen epithelium of calves is affected by liquid feed type [whole milk (WM) or milk replacer (MR)]. Thirty bull calves at the mean age of 5 days were randomly allocated to five experimental groups (six calves/group). Six calves were slaughtered immediately after allocation to the trial (5 days of life), eighteen calves were fed MR and slaughtered at week intervals (on 12, 19, 26 days of life respectively), and six calves were fed WM and slaughtered at the 26 days of life. MCT1 protein abundance and the MCT1 mRNA level were investigated in the dorsal and ventral sack of the rumen. Solid feed intake and short‐chain fatty acids (SCFA) concentration in the rumen fluid increased linearly with calves' age. The amount of the MCT1 protein and mRNA in the dorsal sac of rumen as well as the amount of MCT1 protein in the cranial ventral sac of rumen also increased linearly with calves' age. Calves fed WM had greater solid feed intake in the last week of the study as compared to calves fed MR, but SCFA concentration in the rumen fluid was not different. MCT1 mRNA expression in the cranial dorsal sac of rumen and protein MCT1 expression in both dorsal and ventral cranial sack of the rumen were higher in calves fed WM as compared to calves fed MR. This study confirmed age‐dependent changes of MCT1 expression in the rumen epithelium of newborn calves and showed that its expression might be affected by liquid feed type.