10/10/2016 1 Digestive system 1. Microscopic anatomy of esophagus, stomach, small and large intestine, anus 2. Microscopic anatomy of liver, gall bladder and pancreas 3. Embryonic development of GIT Petr Vaňhara, PhD Department of Histology and Embryology LF MU PVanhara@med.muni.cz General architecture of hollow organs incl. gut tube 1. Mucosa (Tunica mucosa) 2. Submucosa (Tela submucosa) 3. Tunica muscularis externa 4. Serosa/adventitia Donna Myers © 2007 Lumen Serosa/Adventitia Muscularis externa Submucosa Mucosa Four layers Lumen 1 2 3 4 Forstudypurposes 10/10/2016 2 General architecture of hollow organs incl. gut tube Forstudypurposes 10/10/2016 3 Mucosa (Tunica mucosa) - inner layer of gut tube - protective, absorption and resorption - microscopic structure depending on localization - Lamina epithelialis mucosae - Lamina propria mucosae - Lamina muscularis mucosae - Lamina epithelialis mucosae - epithelium type corresponding to function of gut tube - oral cavity, pharynx, esophagus, anus – stratified squamous ep. - stomach, intestine – simple columnar - mucus - secreted by mucosal or submucosal glands (oral cavity, esophagus), secretory epithelium (stomach) or goblet cells (intestine) - Lamina propria mucosae - Layer of mucosal connective tissue – loose collagen - Fenestrated blood capillaries – transport of metabolite (intestine) - mucosal glands in some regions /esophagus) - innervations, immune system - Lamina muscularis mucosae - thin smooth muslce layer - well developer in esophagus, stomach and intestine - small mechanical movements of mucosa facilitating secretion and absorption independently on peristaltic movements. Mucosa (Tunica mucosa) Forstudypurposes 10/10/2016 4 Submucosa (Tela submucosa) Submucosal connective tissue - distinct layer of loose connective tissue - defines shape of mucosa (rugae, plicae) - larger blood and lymph veins nourishing mucosa, muscularis externa and serosa - innervations – nerve plexus - plexus submucosus Meissneri = groups of multipolar neurons and small ganglions, visceral sensory fibers (sympaticus) and fibers and terminal ganglions of parasympaticus (enteric nerve system) - glands – different in different regions; protective function Outer muscular layers (Tunica muscularis externa) - Two concentric, thick layers of smooth muscle, separated by thin layer of connective tissue - Inner – circular, outer – longitudinal (spiral) - Myenteric (Auerbach) plexus - Peristaltic – passage through the gut tube - Local modifications of m.e. - pharyngoesophagal sphincter + external anal sphincter – skeletal muscles - stomach – third - oblique - layer - taenie coli – thickened part of longitudinal layer in colon Circular Longitudinal Forstudypurposes 10/10/2016 5 Serosa/Adventitia (Tunica serosa/adventitia) - outermost layer of gut tube - Serosa - serous membrane of loose connective tissue (Lamina propria serosae) and single layer squamous epithelium (L. epithelialis serosae) - syn. mesothelium, visceral peritoneum - continuous with mesenterium - barrier against various pathogens , antiadhesive properties – intracoelomic movements, immune functions (Ag presentation), ECM production, etc. - Adventitia - some parts of the tube are not covered with epithelium - esophagus in thorax, parts of digestive system in peritoneal cavity in sites of fixation to the walls (duodenum, part of colon, rectum, anal canal) - connective tissue only continuous with connective tissue of the walls S.E. Mutsaers / The International Journal of Biochemistry & Cell Biology 36 (2004) 9–16 Serosa/Adventitia (Tunica serosa/adventitia) 1.4µm Forstudypurposes 10/10/2016 6 Esophagus (Oesophagus) - Mucosa - nonkeratinized stratified squamous epithelium → mechanically protects esophageal tissue - L. propria contains cardial glands (tubular mucinous) and diffuse lymphatic tissue - Submucosa - loose collagen connective tissue, defines shape of mucosa - blood and lymph veins, plexus submucosus Meissneri - submucosal glands (tubular mucinous) - diffuse lymphatic tissue Esophagus (Oesophagus) - Muscularis externa - inner circular and outer longitudinal layer - plexus myentericus Auerbachi - upper third – skeletal muscle, mid third – mixed smooth and skeletal, lower third – smooth muscles only - Adventitia - neck and chest – connects esophagus with surrounding tissue - loose connective tissue - in peritoneal cavity - serosa Forstudypurposes 10/10/2016 7 Cardia of stomach – connection with esophagus Nonkeratinized stratified squamous epithelium → simple columnar epithelium Stomach (Ventriculus, Gaster) - general anatomy of hollow tube - anatomical regions differ also in histologic structure - rugae gastricae (submucosa) Forstudypurposes 10/10/2016 8 Stomach (Ventriculus, Gaster) - Gastric mucosa - simple columnar epithelium - surface epithelium produces mucus (mucinogenic granules, high content of HCO3 -, K+) = protective function - areae gastricae, foveolae gastricae Stomach (Ventriculus, Gaster) - Gastric mucosa - L. propria contains large amount of glands - Gl. cardiacae - Gl. pyloricae - Gl. gastricae propriae Forstudypurposes 10/10/2016 9 Stomach (Ventriculus, Gaster) - Gl. gastricae propriae - glands of fundus and body - simple tubular or branched - 2-4 opens to the gastric pits - four cell types of gl. gastricae propriae chief - most abundant, lower part of body and fundus of the gland - pyramidal shape, basophilic cytoplasm, RER, pepsinogenic granules parietal - neck-body junction - eosinophilic cytoplasm, high numbers of mtch., SER - complex and dynamic ultrastructure - intracellular canals in apical part with microvilli – membrane bound enzyme complexes producing H+ a Cl- (HCl originates extracelullarly) neck cells - cubic, mucinous - capable of regeneration of all cell types in gastric epithelium Stomach(Ventriculus, Gaster) Gl. gastricae propriae Forstudypurposes 10/10/2016 10 Stomach (Ventriculus, Gaster) Gl. gastricae propriae Type Hormone Localization/Function D cells Somatostatin - Stomach, intestine, hepatic and pancreatic ducts EC cells Serotonin - Stomach, gallbladder, intestine - Peristaltics ECL cells Histamin - Stomach - HCl secretion G cells Gastrin - Pars pylorica, duodenum - HCl, pepsin secretion L (EG) cells Enteroglucagon - Stomach, intestine - attenuates secretion of pancreatic enzymes and peristaltics (entero)endocrine - minor, secretion - granules - different cell types with different sensitivity to various histological stainings - secretion of various biologically active compounds - DNES/APUD - GIT chemosensing - see lesson spring semester 2012 - Epithelial tissue Break http://luminaryvisuals.com/ Forstudypurposes 10/10/2016 11 General architecture of hollow organs incl. gut tube Gastroduodenal junction Forstudypurposes 10/10/2016 12 General architecture of the intestine Four basic layers: mucosa, submucosa, muscularis externa, serosa mucosa and submucosa maximise the resorptive area • plicae circulares (Kerckringi) – mucosa + submucosa, ca 800, increase 2-3x, distal region of duodenum • villae (villi intestinales) – mucosa (l. propria + epithelium) 0,5-1,5 mm long, 10- 40/mm2 , 4 000 000, increase 5-10x • microvillae – apical part of enterocytes – 1- 2 µm long, 0,1 µm wide, 100 mil./mm2, increase 20x Small intestine – adaptation to efficient resorption Forstudypurposes 10/10/2016 13 plicae circulares (Kerckringi) – 2-3x villi (villi intestinales) – 5-10x microvilli (striated border) – 20x Small intestine – adaptation to effective resorption Simple columnar epithelium - enterocytes - goblet cells - Paneth cells - enteroendocrine cells - M-cells Intestinal mucosa Crypts of Lieberkühn 200-600x Crypts of Lieberkühn (gl. intestinales) - simple tubular structures of intestinal mucosa, depth 0,3-0,5 mm - pass through l. propria and open to lumen - different cell types - secretion of digestive enzymes - epithelial renewal - enteroendocrine cells - immune response Forstudypurposes 10/10/2016 14 Enterocytes Intestinal mucosa - tall, columnar cells - nucleus located in basis of the cell - apical surface modified- microvilli (3000) + glycocalyx (0,5µm) = striated border (cuticle) - tight intercellular connections, interdigitations Function: - digestion – enzymatic complexes on microvilli membrane - absorption and transport – passive, facilitated i active - lipid uptake - chylomicrons 1µm 0,1µm Forstudypurposes 10/10/2016 15 Microvilli Transportion and resorption Transport of glucose from intestinal lumen to blood stream Na+/K+ ATPase - basolateral surface - concentration gradient Na+ and K+ K+ gradient generates negative membrane potential Na+/glucose symport on apical surface Facilitated diffusion by glucose uniporter (GLUT2) in basolateral membrane Acidification of stomach fluid by parietal cells Apical membrane - H+/K+ ATPase + Cl− a K+ canals Basolateral membrane – anion antiporter HCO3 − and Cl− ions Combined activity of ion channels a cells keeps the electroneutrality and neutral cytoplasmic pH while reaching high extracellular concentration of H+ and Cl− in lumen of stomach http://www.ncbi.nlm.nih.gov/books/NBK21502/ Forstudypurposes 10/10/2016 16 Transportion and resorption water resorbing epithelium water secreting epithelium Jádra F-aktin Mucin v sekrečních granulech - Cylindrical glandular epithelial cells - Apical surface – apocrine/merocrine secretion of mucin - Basal part – RER, GA, nucleus, mitochondria - Mucinogenic granules - see lesson spring semester 2016 - Epithelial tissue Goblet cells Intestinal mucosa Forstudypurposes 10/10/2016 17 Goblet cells Intestinal mucosa Paneth cells Intestinal mucosa - basal part of crypts of Lieberkühn - basophilic cytoplasm - GA located above nucleus - acidophilic (red) granules - immune system - secretion granules contain biologically active substances e.g. lysozym) - influence intestinal microflora Enteroendocrine cells - similar to gastric enteroendocrine cells - regulate pancreatic secretions - homeostatic axis (brain-intestine-adipose tissue) - cholecystokinin, secretin, GIP, motilin, neurocrine peptides etc. Forstudypurposes 10/10/2016 18 M cells (microfold) Intestinal mucosa - epithelial cells above Peyer’s patches and lymphatic nodules - no microvilli - induces immune response - MHCII - antigen presentation to dendritic cells and lymphocytes „Microfold“ Intestinal stem cells Intestinal mucosa - bottom of crypts of Lieberkühn - epithelial renewal (4-5 days) - stem cell niche - tumour transformation Forstudypurposes 10/10/2016 19 L. propria Intestinal mucosa - immune system – GALT - immunologic barrier - Peyer’s patches Brunner’s glands Submucose - gl. duodenale Brunneri - branched tuboalveolar glands, columnar mucinous cells - connective tissue reduced to thin septa between glandular lobules - open to crypts of Lieberkühn Forstudypurposes 10/10/2016 20 Muscularis externa - two layers of smooth muscle (inner circular, outer longitudinal) - plexus myentericus Auerbachi Serosa - loose collagen connective tissue + simple squamous epithelium (mesothelium) Colon - no plicae of Kerckring, villi - muscularis externa – longitudinal layer forms taenie coli - surface serosa forms appendices epiploicae (adipose) Small intestine Colon Forstudypurposes 10/10/2016 21 Colon - absorption of water, electrolytes - deeper crypts of Lieberkühn, no Paneth cells - abundant goblet cells - abundant lymphatic follicles in l. propria (GALT) Forstudypurposes 10/10/2016 22 Apendix - develops from and is connected to caecum 8-10 cm (0,5-1cm) - continuous longitudinal layer of m. externa - lymphatic follicles reaching submucosa - irregular crypts of Lieberkühn with Paneth cells Forstudypurposes 10/10/2016 23 Rectum and anal canal - Pars pelvina - plicae transversae recti - histological architecture identical to colon - Canalis analis - anulus hemorhoidalis – no L. crypts, simple columnar epithelium replaced by stratified squamous epithelium - rich venous plexus - columnae rectales - sinus rectales and valvulae rectales - zona cutanea – typical skin Rectum and anal canal Forstudypurposes 10/10/2016 24 For study purposes 10/10/2016 25 Microscopic anatomy of the gut tube Summary - General architecture of hollow organs and gut tube: mucosa (l. epithelialis m ., l. propria, l. muscularis m.), submucosa, t. muscularis externa, serosa (l. propria s., l. epith. s.), adventitia - Esophagus - structure, epithelium, mucosal and submucosal glands, differences in t. muscularis ext. - Stomach – anatomical and histological structure, mucosa - areae gastricae, foveolae gastricae, gastric glands (pyloricae vs. propriae), localization, ultrastructure and function of gl. gastricae propriae and its cells (chief, parietal, neck, enteroendocrine - Small and large intestine, appendix - anatomical and histological structure, mucosa, glands (crypts of Lieberkühn, Brunner’s glands), cell types of intestinal mucosa, lymphatic system, modifications of intestinal wall - Rectum and anal canal - anatomical and histological structure, mucosa, epithelium, description of associated structures Embryonal development - Development of primitive gut and its derivatives, embryonic flexion, differentiation and characteristics of individual regions and associated organs • Sadler: Langman’s Medical Embryology, 2000 • Ovalle&Nahirney: Netter’s Essential Histology, 2008 • Klika&Vacek: Histologie, 1974 • Ross&Pawlina: Histology (a text and atlas), 2011 • Ross&Romrell: Histology (a text and atlas), 1989 • Berman: Color Atlas of Basic Histology • Ústav histologie & embryologie LF MU, www.med.muni.cz/histol/histolc.html • Čech S., Horký D., Sedláčková M.: Přehled embryologie člověka, Brno, LF MU, 2011 • Horký D., Čech S.: Mikroskopická anatomie, Brno, LF MU, 2011 • About.com Health's Disease and Condition, Donna Myers © 2007 • Mutsaers SE, The mesothelial cell, The International Journal of Biochemistry & Cell Biology, 2004, 36(1):9-16 • A.D.A.M. Education /University of Maryland Medical Center (UMMC) http://www.umm.edu/; http://www.adameducation.com/index.aspx • Science Photo Library • Barker N, Bartfeld S, Clevers H. Tissue-resident adult stem cell populations of rapidly self-renewing organs. Cell Stem Cell. 2010 Dec 3;7(6):656-70. • Mills JC, Shivdasani RA. Gastric epithelial stem cells. Gastroenterology. 2011 Feb;140(2):412-24. • Ohno H. M cells hold the key to gut immunity. Riken Research Frontlines. 2010. http://www.rikenresearch.riken.jp/eng/frontline/6346 • Kosinski C et al. Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors. PNAS 2007;104:15418-15423 • Knoblich JA. Asymmetric cell division during animal development. Nature Reviews Molecular Cell Biology 2001, 2, 11-20 • van der Flier LG, Clevers H. Stem Cells, Self-Renewal, and Differentiation in the Intestinal Epithelium. Annu. Rev. Physiol. 2009.71:241-260 • Southern Illionis University, http://www.siumed.edu/~dking2/erg/GI015b.htm Study materials References Forstudypurposes