Morphology and Mucin Histochemistry of the Gastrointestinal Tracts of Three Insectivorous Mammals: Acomys Spinosissimus, Crocidura Cyanea and Amblysomus Hottentotus
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Morphology and Mucin Histochemistry of the Gastrointestinal Tracts of Three Insectivorous Mammals: Acomys spinosissimus, Crocidura cyanea and Amblysomus hottentotus by Julia Boonzaier Thesis presented in partial fulfilment of the requirements for the degree of Master of Medical Science in the Faculty of Health Sciences at Stellenbosch University. Supervisor: Dr. Sanet H. Kotzé Department of Biomedical Sciences Faculty of Health Sciences University of Stellenbosch Co-supervisor: Dr. Elizabeth L. van der Merwe Department of Human Biology Faculty of Health Sciences University of Cape Town March 2012 Page | I Stellenbosch University http://scholar.sun.ac.za DECLARATION By submitting this thesis electronically, I declare that the entirety of the work contained therein is my own, original work, that I am the sole author thereof, that reproduction and publication thereof by Stellenbosch University will not infringe any third party rights and that I have not previously in its entirety or in part submitted it for obtaining any qualification. Date: 08 December 2011 Copyright © 2012 Stellenbosch University All rights reserved Page | II Stellenbosch University http://scholar.sun.ac.za ABSTRACT The gastrointestinal morphology and the distribution of the different types of mucin secreting goblet cells were investigated in three mammalian insectivorous species, namely A. spinosissimus, C. cyanea and A. hottentotus. The aim of the study was to provide a comprehensive morphological comparison between the different species. Another aim was to illustrate and compare the distribution of mucins (neutral, sulfo- and sialomucins) in the gastrointestinal tracts (GITs) of these species, in order to better understand the quality of the biofilm in the GIT. Mucins secreted onto the surface of the GIT have an effect on the colonisation of microflora in the mucosal layer, constructing a biofilm which protects the GIT surface from opportunistic pathogens. The shape, proportional length, and proportional surface areas of the different gastrointestinal regions were recorded and compared in the three species. Histochemical staining methods were used to detect and to distinguish between neutral, sulfo- and sialomucins. The number of goblet cells in the GIT containing each of the above mucins in the epithelium lining the surface or crypts was quantified, and the data expressed as the number of neutral, sulfo- or sialomucin containing goblet cells per mm2 of the surface or crypt epithelium. In all three species the stomach was uncompartmentalised. The internal aspect of the stomach in A. spinosissimus was hemi-glandular, containing stratified squamous epithelium in the fundus, with glandular epithelium in the body and pyloric region. However, C. cyanea and A. hottentotus had wholly glandular stomachs. A. spinosissimus was the only species studied that had a caecum which demonstrated transverse mucosal folds and V-shaped mucosal folds in the proximal colon. Both C. cyanea and A. hottentotus had villi up to the distal part of the GIT. Longitudinal mucosal folds were present in the distal colon. The GITs of both C. cyanea and A. hottentotus showed little morphological differentiation namely a simple, glandular stomach and the lack of a caecum. Mixed (neutral and acid) mucins and mixed acid (sulfo- and sialomucins) mucin secreting goblet cells were prominent mucin cell types in all three mammalian insectivorous species. Despite these general similarities, marked differences were observed in the qualitative expression and distribution of the three types of mucins throughout the GIT. The overall similarity between the three insectivores and other distantly related mammalian species suggests that mixed mucin secreting goblet cell types are prominent contributors to the Page | III Stellenbosch University http://scholar.sun.ac.za maintenance of the intestinal biofilm in the majority of mammals, irrespective of their diet or taxonomy. Page | IV Stellenbosch University http://scholar.sun.ac.za OPSOMMING Die bestudering van die morfologie van die spysverteringskanaal (SVK) en die verspreiding van die verskillende musien produserende bekerselle was in drie insek- etende soogdier spesies uitgevoer, naamlik in A. spinosissimus, C. cyanea en A. hottentotus. Die doel van die studie was om „n omvattende morfologiese vergelyking te maak tussen die drie spesies, sowel as om die verspreiding van die verskillende musiene te beskryf in die SVK. Kennis van die verspreiding van die verskillende tipes musiene (neutral, sulfaat en nie-sulfaat bevattende musiene) kan moontlik inligting verskaf aangaande die kwaliteit van die biofilm in the SVK. Die laasgenoemde musiene wat gesekreteer word op die oppervlak van die SVK, bepaal die kolonisasie van die mikroflora in die mukosale laag wat „n biofilm vorm en die SVK beskerm teen patogene. Die vorm, proportionele lengte en proportionele oppervlaks areas van die verskillende SVK gebiede is opgeteken, waarna dit vergelyk is tussen die drie insektivore spesies. Histochemiese kleurings tegnieke is gebruik om die musiene waar te neem en om te onderskei tussen die neutraal, sulfaat en nie-sulfaat bevattende musiene. Die aantal beker selle wat elk van die bogenoemde musiene bevat het, is getel in die oppervlaks epiteel- en kript areas van die SVK. Hierdie data is weergegee as die aantal neutraal, sulfaat en nie- sulfaat bevattende beker selle per oppervlaks epiteel- of kript area (mm2). Die vorm van die maag in al drie spesies was eenvoudig en nie gekompartementaliseer nie. Die interne aspek van die maag in A. spinosissimus het meerlagige plaveisel epiteel in die fundus gehad en klieragtige epiteel in die liggaam en pilorus gedeeltes. Daarbenewens het C. cyanea en A. hottentotus slegs klieragtige epiteel in die maag gehad. A. spinosissimus was die enigste spesie in hierdie studie wat „n sekum gehad het met dwars voue, asook V-vormige mukosale voue in die proksimale kolon. Beide C. cyanea en A. hottentotus het villi tot in die distale gedeelte van die SVK gehad. Longitudinale mukosale voue was teenwoordig in die distale gedeelte van die kolon. Die SVK van beide C. cyanea en A. hottentotus het min morfologiese differensiasie getoon deurdat die spesies „n eenvoudige, klieragtige maag gehad het en geen sekum nie. Gemenge (neutral en suur) musiene asook gemengde suur (sulfaat en nie-sulfaat bevattende) musiene was die dominante musien tipes in the SVK van al drie insek-etende soogdier spesies. Ten spyte van die algemene ooreenkomste, was daar merkwaardige verskille in die getalle en verspreiding van die verskillende musiene in die SVK. Die algemene ooreenkomste tussen die drie insektivore soogdier spesies met soogdiere van Page | V Stellenbosch University http://scholar.sun.ac.za ander families, stel voor dat die gemende musien sekreterende beker selle „n prominente bydrae maak tot die onderhoud van die biofilm in the SVK in die meerderheid van soogdiere, ongeag van hul dieet of spesie klassifikasie. Page | VI Stellenbosch University http://scholar.sun.ac.za AKNOWLEDGEMENTS This report would not have been possible without the essential and gracious support of many individuals. A special thanks to my supervisors, Dr. S. H. Kotzé and Dr. E. L. van der Merwe for all their effort and support during the writing of this document. Your patience, kindness, as well as your academic experience has been invaluable to me. I would like to thank Prof. Nigel Bennett for donating the specimens that were used in this study to the Division of Anatomy and Histology. Thank you for all your time and effort in the capturing of the animals and ensuring that the specimens were handled correctly. I am extremely grateful to Prof. Martin Kidd for the statistical analysis of all the data, and also for assistance in the interpretation and understanding thereof. Thank you to Mr Reggie Williams and Miss Zandré Fourie, at the University of Stellenbosch and Morea Petersen at the University of Cape Town for technical support with the histochemical techniques. Thank you to the Division of Anatomy and Histology (Department of Biomedical Sciences, Faculty of Health Sciences), and the University of Stellenbosch for financial assistance and for the opportunity to attend the ASSA conference. Thank you to the Harry Crossley Foundation for funding this project. Without this financial support this project would not have been possible. I would also like to acknowledge the following authors for the use of their photographs and illustrations in my thesis: Mills & Hess (1997); Stevens & Hume (1998); Shirazi et al. (2000); Kierszenbaum (2002); Young et al. (2006); Brockhausen et al. (2009) and McGuckin et al. (2009). I am grateful to my surrogate family (Frik Fourie, Erika Fourie, Elmarie Fourie), Pieter Fourie, and all of my friends and everyone at Anatomy and Histology for their help and moral support. I would especially like to thank Elsje-Marie Geldenhuys, Mandi Alblas and Linda Greyling for their support. Finally, I am forever indebted to my parents for their understanding, endless patience and encouragement when it was most required. Page | VII Stellenbosch University http://scholar.sun.ac.za TABLE OF CONTENTS DECLARATION ..................................................................................................................... II ABSTRACT .......................................................................................................................... III OPSOMMING ......................................................................................................................