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Research Article
 

Gut Endocrine Cells in the Stomach of Sunda Porcupines (Hystrix javanica)



Teguh Budipitojo, Yuda Heru Fibrianto and Guntari Titik Mulyani
 
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ABSTRACT

Background and Objective: The regional distribution and relative frequency of gastrointestinal endocrine cells has been found to vary with the animal species and feeding habits. This study aimed to analyze the existence and distribution of immunoreactive endocrine cells to chromogranin, ghrelin, somatostatin, serotonin and gastrin in the gastric mucosa of sunda porcupine (Hystrix javanica ) using immunohistochemistry. Materials and Methods: Five adult sunda porcupines were used without sexual distinction. The presences of endocrine cells were visualized by ABC immunohistochemical methods using five types of polyclonal antisera, specific for chromogranin, ghrelin, somatostatin, serotonin and gastrin. Results: In the stomach of sunda porcupine, endocrine cells immunoreactive for all of these markers were observed. The chromogranin-immunoreactive cells were found with greatest frequency, in all stomach regions. In all regions of the stomach, endocrine cells secreting ghrelin were also detected in weaker frequency compare with chromogranin. The somatostatin immunoreactive cells were detected rare in the gastric and fundic regions, serotonin cells were found rare in fundic but few in pyloric regions, while gastrin cells were detected abundance in pyloric region of stomach of the sunda porcupines. Gastrin positive cell was also detected in the duodenal part of sunda porcupine. The chromogranin-positive cells were the most numerous, being more prevalent in the stomach glands. The other endocrine cells were identified in smaller numbers, some of them located in all regions or in specific regions. The finding of these cell types in the mucosal stomach confirms their preferential location in the final portions of the principal regions of the stomach and suggests control by feedback of its functions. Conclusion: In conclusion, the present study show for the first time the immunolocalization of chromogranin, ghrelin and gastrin immunoreactive cells in the stomach of sunda porcupine. However, in contrast with previous finding of crested porcupine and other rodents, the present result found serotonin immunoreactive cells in the fundic and pyloric gland regions and somatostatin immunoreactive cells in the gastric and fundic gland regions of sunda porcupine stomach. The differences may was caused by feed types, feeding habits and geographical locations of the species.

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  How to cite this article:

Teguh Budipitojo, Yuda Heru Fibrianto and Guntari Titik Mulyani, 2016. Gut Endocrine Cells in the Stomach of Sunda Porcupines (Hystrix javanica). Asian Journal of Animal Sciences, 10: 268-272.

DOI: 10.3923/ajas.2016.268.272

URL: https://scialert.net/abstract/?doi=ajas.2016.268.272
 
Received: May 16, 2016; Accepted: July 01, 2016; Published: August 15, 2016



INTRODUCTION

Gastrointestinal endocrine cells were dispersed throughout the epithelia and gastric glands of the alimentary tract. These cells produce many biologically active polypeptides and amines, some of them hormones1. These hormones have important functions in the overall regulation of the digestive process, such as secretion of intestinal and associated glands, gut motility, nutrient absorption and intestinal blood flow2. The role of endocrine cells and their respective regulator peptides has attracted the interest of many investigators, who have described them in the gastrintestinal tract (GIT) of other vertebrates, such as amphibia3, fish4, birds5 and mammals6, seeking to establish the morphological role of these cells and respective peptides responsible for regulating numerous body functions.

The regional distribution and relative frequency of gastrointestinal endocrine cells has been found to vary with the animal species and feeding habits2. Fourteen types of endocrine cells have been demonstrated in the gastrointestinal tract of Tree Shrew7 and Babirusa8 by immunohistochemistry. Six types of endocrine cells have been shown in the malayan pangolin, manis javanica9 and five in the barking deer10. These studies reveal interspecific differences and suggest correlation between endocrine cell distribution and feeding habits. This study aimed to analyze the existence and distribution of immunoreactive endocrine cells to chromogranin, ghrelin, somatostatin, serotonin and gastrin in the gastric mucosa of sunda porcupine (Hystrix javanica) using immunohistochemistry.

MATERIALS AND METHODS

Five stomachs of adult sunda porcupines, Hystrix javanica, about 67 cm in length, were purchased from a merchant in Tawangmangu, Central java, Indonesia and were used as samples. Stomach tissues of Hystrix javanica were fixed for 24 h in Bouin's solution, dehydrate in ethanol, cleared in xylene and embbeded in parrafin.

Stomach sections were cut serially in 4-5 μm thickenesses and stained by hematoxylin and eosin (HE) for conventional histological evaluation. For immunohistochemical staining of avidin-biotin peroxidase complex (ABC) methods11 tissue sections were deparaffinized in xylene and rehydrated in decreasing series of ethanol concentrations. After washing in Phosphate Buffered Saline (PBS), endogenous peroxidase activity was blocked by incubating the section in H2O2 in methanol for 15 min. Primary antibodies of porcine chromogranin (Lot 8808014; Incstar, Stilwater, OK, USA) at 1:5,000 dilutions, ghrelin (bs-0467R; Bioss Inc. Massachusetts, USA) at 1:1000 dilution, serotonin (Sero-2-3; J Nishitsutsuji, Japan) at 1:5,000 dilution, gastrin (GP-1304; Yanaihara Institute, Japan) at 1:10,000 dilution and somatostatin (20H2T; Incstar, Stilwater, OK, USA) at 1:5,000 dilution were applied overnight at 4°C. To prevent non spesific staining, sections were incubate with nomal goat serum before incubation with the primary antibodies. The secondary antibody was applied for 45 min at room temperature. The immunoreactive site were visualized by tris-HCL buffer containing 3,3 diaminobenzidine tetrahydrocloride. Section were counterstained with mayer hematoxylin and examined with a conventional light microscope and photomicrographs were taken with digital camera.

The specificity of the immunohistochemical staining was confirmed by the replacement of primary antibody with normal rabbit serum. The semiquantitatif number of positif endocrine cell were graded subjectively into 4 classes as rare, few, moderate and numerous.

RESULTS

The present study revealed five kinds of endocrine cells immunoreactive for chromogranin, ghrelin, somatostatin, serotonin and gastrin in the gastric mucosa of sunda porcupine (Hystrix javanica). The chromogranin-positive cells were the most numerous, being more prevalent in the stomach glands. The other endocrine cells were identified in smaller numbers, some of them located in all regions or in specific regions.

The chromogranin immunoreactive cells were found rare, few and numerous in the gastric, fundic and pyloric gland regions, respectively (Fig. 1a-c). In all regions of the stomach, endocrine cells secreting ghrelin were detected rare, a weaker frequency compare with chromogranin (Fig. 1d-f).

The somatostatin immunoreactive cells were detected rare in the gastric and fundic regions (Fig. 2a, b), serotonin immunoreactive cells were found rare in fundic but few in pyloric regions (Fig. 2c, d), while gastrin immunoreactive cells were detected numerous in pyloric region of stomach of the sunda porcupines (Fig. 2e). Gastrin positive cell was also detected in the duodenal part of sunda porcupine, individually (Fig. 1f).

Image for - Gut Endocrine Cells in the Stomach of Sunda Porcupines (Hystrix javanica)
Fig. 1(a-f):
Immonolocalization of chromogranin and ghrelin positive cells in the gastric, fundic and pyloric regions of the stomach of sunda porcupine (Hystrix javanica). As indicated by arrows, chromogranin cells were detected rare, few and numerous in the (a) Gastric, (b) Fundic, (c) Pyloric gland regions, respectively, while ghrelin cells were detected rare in (d) Gastric, (e) Fundic and (f) Pyloric gland regions of stomach of the sunda porcupines

Image for - Gut Endocrine Cells in the Stomach of Sunda Porcupines (Hystrix javanica)
Fig. 2(a-f):
Immonolocalization of somatostatin, serotonin and gastrin positive cells in the stomach of sunda porcupine (Hystrix javanica). As indicated by arrows, somatostatin cells were detected rare in the (a) Gastric, (b) Fundic gland regions, serotonin cells were found rare in (c) Fundic but few in (d) Pyloric gland regions, while gastrin cells were detected numerous in (e) Pyloric gland region of stomach of the sunda porcupines and (f) Gastrin positive cell was also detected in the duodenal part of sunda porcupine

DISCUSSION

Chromogranin has been used as a marker of endocrine cells as it is found in a wide spectrum of gut endocrine cells in mammals12 and was detected in all of stomach gland regions of babirusa8 and malayan pangolin9. In agreement with these previous findings, the present study found the presences of chromogranin immunoreactive cells in all gastric gland regions of sunda porcupine.

Ghrelin is a peptide hormone produced in the gastrointestinal tract which functions as a neuropeptide in the central nervous system13. Besides regulating appetite, ghrelin also plays a significant role in regulating the distribution and rate of use of energy14. Ghrelin immunoreactive cells are found mainly in the stomach15. In rodents, ghrelin immunoreactive cells were observed in all regions of the gastrointestinal tract and were most abundance in the mucosal layer of fundic gland region16. In agreement with these previous findings, the present study found ghrelin immunoreactive cells in all gastric gland regions of sunda porcupine without any differences on the density between gland regions.

Serotonin is secreted by the enterocromafin cells, widely distributed in the nervous system and in the gastro-entero-pancreatic endocrine cells17, where it is used to regulate intestinal movements18. Serotonin immunoreactive cells were detected in the whole gastrointestinal tract of rat19, mouse20 and porcupine21. In the crested porcupine, serotonin immunoreactive cells were detected throughout the whole gastrointestinal tract and showed the highest frequencies in the cardiac region of stomach and duodenum21,22. In contrast with previous finding in crested porcupine and other rodents, the present result found serotonin immunoreactive cells in the fundic and pyloric gland regions of sunda porcupine stomach but not in the gastric gland region.

The somatostatin is a neuroendocrine hormone which was first isolated from the hypothalamus of the sheep23 and plays important inhibition roles on the secretion of other neuroendocrine hormones24. Somatostatin immunoreactive cells were found predominantly in the pylorus gland region of manchurian chipmunk25 and gerbil26 stomach. In contrast with previous finding, somatostatin immunoreactive cells were not detected in the stomach of the crested porcupine22. In the present study, however, somatostatin immunoreactive cells were found in the gastric and fundic gland regions of sunda porcupine stomach but not in the pyloric gland region.

Gastrin is a peptide hormone produced by G cells of the pyloric antrum of the stomach and duodenum27. This hormone acts by increasing the force of antrum contractions and the constriction of the pyloric sphincter28. In agreement with ostrich29 and malayan pangolin9, the present study found gastrin immunoreactive cells in the gastric glands of the pyloric gland region and duodenum of sunda porcupine (Hystrix javanica).

CONCLUSION

The present study show for the 1st time the immunolocalization of chromogranin, ghrelin and gastrin immunoreactive cells in the stomach of sunda porcupine. However, in contrast with previous finding of crested porcupine and other rodents, the present result found serotonin immunoreactive cells in the fundic and pyloric gland regions and somatostatin immunoreactive cells in the gastric and fundic gland regions of sunda porcupine stomach. The differences may caused by feed types, feeding habits and geographical locations of the species.

SIGNIFICANT STATEMENT

The present study show for the first time the immunolocalization of chromogranin, ghrelin and gastrin immunoreactive cells in the stomach of porcupine, especially sunda porcupine (Hystrix javanica) which is endemic to Indonesia. In the previous report on crested procupine (Hystrix cristata) which is endemic to Italy, Sicily, North Africa and sub-Saharan Africa. Timurkaan et al.21 and Yaman et al.22 showed the presence of serotonin and somatostatin in the gastrointestinal tract. However, in contrast with previous finding of crested porcupine and other rodents, the present result found serotonin immunoreactive cells in the fundic and pyloric gland regions and somatostatin immunoreactive cells in the gastric and fundic gland regions of sunda porcupine stomach.

ACKNOWLEDGMENT

This study was full supported by the grant for scientific research (No. 38/LPPMUGM/2015) from the Directorate General of Higher Education (DIKTI), Ministry of Research, Technology and Higher Education of Indonesia.

REFERENCES

1:  Schonhoff, S.E., M. Giel-Moloney and A.B. Leiter, 2004. Minireview: Development and differentiation of gut endocrine cells. Endocrinology, 145: 2639-2644.
CrossRef  |  Direct Link  |  

2:  Gunawardene, A.R., B.M. Corfe and C.A. Staton, 2011. Classification and functions of enteroendocrine cells of the lower gastrointestinal tract. Int. J. Exp. Pathol., 92: 219-231.
CrossRef  |  Direct Link  |  

3:  Xie, Z., D. Guo, C. Lu, B. Li and L. Shan, 2012. Argentaffine cells in the digestive tract of hibernating and non-hibernating in Chinese fire-bellied newt (Cynops orientalis). Int. J. Morphol., 30: 1389-1394.
Direct Link  |  

4:  Vieira-Lopes, D.A., N.L. Pinheiro, A. Sales, A. Ventura, F.G. Araujo, I.D. Gomes and A.A. Nascimento, 2013. Immunohistochemical study of the digestive tract of Oligosarcus hepsetus. World J. Gastroenterol., 19: 1919-1929.
CrossRef  |  Direct Link  |  

5:  Yang, J., L. Zhang, X. Li, L. Zhang, X. Liu and K. Peng, 2012. An immunohistochemical study on the distribution of endocrine cells in the digestive tract of gray goose (Anser anser). Turk. J. Vet. Anim. Sci., 36: 373-379.
Direct Link  |  

6:  Furness, J.B., L.R. Rivera, H.J. Cho, D.M. Bravo and B. Callaghan, 2013. The gut as a sensory organ. Nat. Rev. Gastroenterol. Hepatol., 10: 729-740.
CrossRef  |  Direct Link  |  

7:  Yamada, J., M. Tauchi, W. Rerkamnuaychoke, H. Endo and N. Chungsamarnyart et al., 1999. Immunohistochemical survey of the gut endocrine cells in the common tree shrew (Tupaia belangeri). J. Vet. Med. Sci., 61: 761-767.
PubMed  |  Direct Link  |  

8:  Agungpriyono, S., A.A. Macdonald, K.Y. Leus, N. Kitamura and I.K. Adnyane et al., 2000. Immunohistochemical study on the distribution of endocrine cells in the gastrointestinal tract of the babirusa, Babyrousa babyrussa (Suidae). Anat. Histol. Embryol., 29: 173-178.
CrossRef  |  PubMed  |  Direct Link  |  

9:  Nisa, C., N. Kitamura, M. Sasaki, S. Agungpriyono and C. Choliq et al., 2005. Immunohistochemical study on the distribution and relative frequency of endocrine cells in the stomach of the Malayan Pangolin, Manis javanica. Anat. Histol. Embryol., 34: 373-378.
CrossRef  |  PubMed  |  Direct Link  |  

10:  Adnyane, I.K.M., A.B. Zuki, M.M. Noordin and S. Agungpriyono, 2011. Immunohistochemical study of endocrine cells in the gastrointestinal tract of the barking deer, Muntiacus muntjak. Anatomia Histol. Embryol., 40: 365-374.
CrossRef  |  Direct Link  |  

11:  Dabbs, D.J., 2014. Diagnostic Immunohistochemistry. 3rd Edn., Churchill Livingstone Elsevier, Philadelphia, PA., USA

12:  El-Salhy, M., O.H. Gilja, D. Gundersen, J.G. Hatlebakk and T. Hausken, 2014. Duodenal chromogranin a cell density as a biomarker for the diagnosis of irritable bowel syndrome. Gastroenterol. Res. Pract.
CrossRef  |  Direct Link  |  

13:  Burger, K.S. and L.A. Berner, 2014. A functional neuroimaging review of obesity, appetitive hormones and ingestive behavior. Physiol. Behav., 136: 121-127.
CrossRef  |  Direct Link  |  

14:  Dickson, S.L., E. Egecioglu, S. Landgren, K.P. Skibicka, J.A. Engel and E. Jerlhag, 2011. The role of the central ghrelin system in reward from food and chemical drugs. Mol. Cell. Endocrinol., 340: 80-87.
CrossRef  |  Direct Link  |  

15:  Hamza, S., I. Vulkova, M. Gulubova and D. Sivrev, 2014. Characteristics of ghrelinpositive cells of the stomach in the rat. Trak. J. Sci., 12: 141-143.
Direct Link  |  

16:  Sakata, I. and T. Sakai, 2010. Ghrelin cells in the gastrointestinal tract. Int. J. Peptides.
CrossRef  |  Direct Link  |  

17:  Richter, G., F. Stockmann, J.M. Conlon and W. Creutzfeldt, 1986. Serotonin release into blood after food and pentagastrin: Studies in healthy subjects and in patients with metastatic carcinoid tumors. Gastroenterology, 91: 612-618.
CrossRef  |  Direct Link  |  

18:  Bornstein, J.C., 2012. Serotonin in the gut: What does it do? Front. Neurosci., Vol. 6.
CrossRef  |  Direct Link  |  

19:  Singh, P., S.R. Dutta and D. Guha, 2015. Gastric mucosal protection by aegle marmelos against gastric mucosal damage: Role of enterochromaffin cell and serotonin. Saudi J. Gastroenterol., 21: 35-42.
Direct Link  |  

20:  Yano, J.M., K. Yu, G.P. Donaldson, G.G. Shastri and P. Ann et al., 2015. Indigenous bacteria from the gut microbiota regulate host serotonin biosynthesis. Cell, 161: 264-276.
CrossRef  |  Direct Link  |  

21:  Timurkaan, S., M. Karan and A. Aydin, 2005. Immunohistochemical study of the distribution of serotonin in the gastrointestinal tract of the porcupines (Hystrix cristata). Rev. Vet. Med., 156: 533-536.
Direct Link  |  

22:  Yaman, M., B.G. Tarakci, A. Bayrakdar, O. Atalar and O. Dabak, 2007. Immunohistochemical study of gastrointestinal endocrine cells in the porcupine (Hystrix cristata). Rev. Med. Vet., 158: 196-200.
Direct Link  |  

23:  Brazeau, P., W. Vale, R. Burgus, N. Ling, M. Butcher, J. Rivier and R. Guillemin, 1973. Hypothalamic polypeptide that inhibits the secretion of immunoreactive pituitary growth hormone. Science, 179: 77-79.
CrossRef  |  PubMed  |  Direct Link  |  

24:  Wolin, E.M., 2012. The expanding role of somatostatin analogs in the management of neuroendocrine tumors. Gastrointest Cancer Res., 5: 161-168.
Direct Link  |  

25:  Lee, H.S., S.K. Ku and J.H. Lee, 1998. Localization of endocrine cells in the gastrointestinal tract of the Manchurian chipmunk, Tamias sibiricus barberi. Korean J. Biol. Sci., 2: 395-401.
CrossRef  |  Direct Link  |  

26:  Lee, H.J., H.S. Lee, S.K. Ku, K.D. Park and K.S. Kim, 2000. Immunohistochemical study of the gastrointestinal endocrine cells in the Mongolian Gerbils, Meriones unguiculatus. Korean J. Vet. Res., 40: 653-660.
Direct Link  |  

27:  Kasajima, A., F. Fujishima, T. Morikawa, S. Kawasaki and S. Konosu-Fukaya et al., 2015. G-cell hyperplasia of the stomach induces ECL-cell proliferation in the pyloric glands in a paracrinal manner. Pathol. Int., 65: 259-263.
CrossRef  |  Direct Link  |  

28:  Rau, T.T., A. Sonst, A. Rogler, G. Burnat and H. Neumann et al., 2013. Gastrin mediated down regulation of ghrelin and its pathophysiological role in atrophic gastritis. J. Physiol. Pharmacol., 64: 719-725.
PubMed  |  Direct Link  |  

29:  Duritis, I., A. Mugurevics and L. Mancevica, 2013. The distribution of gastrin, somatostatin and glucagon immunoreactive (IR) cells in ostrich stomach during the pre‐and post‐hatching period. Anat. Histol. Embryol., 42: 362-368.
CrossRef  |  Direct Link  |  

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