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Samenvatting Celbiologie en Algemene weefselleer
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Samenvatting van Celbiologie en Algemene weefselleer Professor Wim Van Den Broeck UGent Diergeneeskunde
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CELBIOLOGIE EN ALGEMENEWEEFSELLEER
Wim Van Den Broeck
1e Bachelor Diergeneeskunde Ugent 2021-‘22
,Inhoud
Inleidende begrippen .............................................................................................................................. 7
Optische principes ................................................................................................................................... 7
Microscopie – eenheden ..................................................................................................................... 7
Oplossend vermogen en contrast ....................................................................................................... 8
De lichtmicroscoop .................................................................................................................................. 8
Gewone lichtmicroscoop ..................................................................................................................... 8
Fasecontrastmicroscoop ..................................................................................................................... 8
Donkerveldmicroscoop ....................................................................................................................... 8
Polarisatiemicroscoop ......................................................................................................................... 8
Fluorescentiemicroscoop .................................................................................................................... 8
De elektronenmicroscoop ....................................................................................................................... 9
Algemeen............................................................................................................................................. 9
Transmissie elektronenmicroscoop (TEM) .......................................................................................... 9
Scanning elektronenmicroscoop (SEM) .............................................................................................. 9
Methodologie .......................................................................................................................................... 9
Paraffinemethode (LM) ....................................................................................................................... 9
Vriessnedemethode (LM en EM)....................................................................................................... 10
Transmissie elektronenmicroscopie.................................................................................................. 10
Scanning elektronenmicroscopie ...................................................................................................... 10
Weefselcultuur .................................................................................................................................. 10
Morfologisch onderzoek ................................................................................................................... 10
Histochemie....................................................................................................................................... 11
Enzymhistochemie ............................................................................................................................ 11
Autoradiografie ................................................................................................................................. 11
Immunohistologie ............................................................................................................................. 11
Algemene structuur van de cel.............................................................................................................. 12
Optische waarneming ....................................................................................................................... 12
Structuur van de cel .......................................................................................................................... 13
Specialisatie van de cel ...................................................................................................................... 15
De celmembraan – membraanstructuren............................................................................................. 17
“unit membrane” .............................................................................................................................. 17
Lipide-proteïne model ....................................................................................................................... 17
Vloeibaar lipide-proteïne model ................................................................................................... 17
Samenstelling celmembraan ......................................................................................................... 17
De celmantel...................................................................................................................................... 18
1
, Functies plasmalemma en celmantel ................................................................................................ 19
Contactdifferentiaties: differentiaties celmembraan ivm contact tussen de cellen......................... 19
Oppervlaktetoename ivm resorptie en secretie ............................................................................... 20
Tijdelijke membraanmodificaties/vormveranderingen v/h plasmalemma ...................................... 20
Opname en afgave ........................................................................................................................ 20
Beweging ....................................................................................................................................... 20
De kern – nucleus – karyon ................................................................................................................... 21
De celcyclus ....................................................................................................................................... 21
Algemene kenmerken – interfase ..................................................................................................... 21
Kern envelop – interfase ............................................................................................................... 22
Nucleoplasma – interfase .............................................................................................................. 22
Chromatine – interfase.................................................................................................................. 23
Chromatine – metafase ..................................................................................................................... 24
Nucleolus ....................................................................................................................................... 25
Celdeling ............................................................................................................................................ 26
De celcyclus ................................................................................................................................... 26
Mitose – kenmerken ..................................................................................................................... 27
Mitose – fasen ............................................................................................................................... 27
Meiose ........................................................................................................................................... 28
Mitose – meiose ............................................................................................................................ 29
De ribosomen en het endoplasmatisch reticulum ................................................................................ 29
Ribosomen ......................................................................................................................................... 29
Morfologie ..................................................................................................................................... 29
Samenstelling ................................................................................................................................ 29
Functie ........................................................................................................................................... 30
Afbraak eiwitten ............................................................................................................................ 30
Endoplasmatisch reticulum ............................................................................................................... 30
Morfologie ..................................................................................................................................... 30
Ruw endoplasmatisch reticulum RER ............................................................................................ 30
Glad endoplasmatisch reticulum GER ........................................................................................... 31
Het golgi-complex.................................................................................................................................. 31
Morfologie ......................................................................................................................................... 31
Functie ............................................................................................................................................... 31
Endosomen ........................................................................................................................................ 32
Ontstaan ........................................................................................................................................ 32
Kenmerken .................................................................................................................................... 32
2
, Functie ........................................................................................................................................... 32
Lysosomen ............................................................................................................................................. 33
Ontstaan ............................................................................................................................................ 33
Morfologie ......................................................................................................................................... 33
Functie ............................................................................................................................................... 33
Disfunctie........................................................................................................................................... 34
Peroxisomen .......................................................................................................................................... 34
Morfologie ......................................................................................................................................... 34
Functie ............................................................................................................................................... 34
Mitochondriën....................................................................................................................................... 35
Morfologie ......................................................................................................................................... 35
Aantoonbaarheid............................................................................................................................... 35
Functie ............................................................................................................................................... 36
Oorsprong, replicatie......................................................................................................................... 36
Apoptose ........................................................................................................................................... 36
Cytoskelet .............................................................................................................................................. 36
Inleiding ............................................................................................................................................. 36
Intermediaire filamenten .................................................................................................................. 37
Microtubuli ........................................................................................................................................ 39
Microfilamenten ................................................................................................................................ 42
Celinclusies – paraplasma ..................................................................................................................... 43
Indeling .............................................................................................................................................. 43
Secretorische activiteit ...................................................................................................................... 43
Reservemateriaal............................................................................................................................... 44
Glycogeen ...................................................................................................................................... 44
Vetvacuolen ................................................................................................................................... 44
Eiwitstapeling ................................................................................................................................ 44
Pigmentinclusies ................................................................................................................................ 44
Endogene pigmenten .................................................................................................................... 44
Exogene pigmenten ....................................................................................................................... 45
Celdood – apoptose .............................................................................................................................. 46
Accidentele vs gereguleerde celdood ............................................................................................... 46
Morfologische kenmerken ................................................................................................................ 46
Apoptose vs necrose ......................................................................................................................... 47
Regulated cell death .......................................................................................................................... 48
Genetische basis van celdood ........................................................................................................... 48
3
, Caspases ........................................................................................................................................ 48
Caspase activatie ........................................................................................................................... 48
Apoptose: intrinsiek en extrinsieke route ......................................................................................... 49
Intrinsiek: mitochondriaal mechanisme ........................................................................................ 49
Intrinsieke apoptose (mitochondriaal mechanisme) .................................................................... 49
Apoptose mechanismen.................................................................................................................... 49
Extrinsiek: receptor-ligand gemedieerde apoptose .......................................................................... 49
Extrinsieke apoptose (receptor gemedieerd mechanisme) .............................................................. 50
Apoptose inhibitie ............................................................................................................................. 50
Histologisch aantonen van apoptose ................................................................................................ 50
Epitheelweefsel ..................................................................................................................................... 51
Eigenschappen................................................................................................................................... 51
Bedekkingsepitheel ........................................................................................................................... 55
Klierepitheel ...................................................................................................................................... 59
Regeneratie ....................................................................................................................................... 61
Het gewoon bindweefsel....................................................................................................................... 62
Structuur – grondstof ........................................................................................................................ 62
Structuur-vezels................................................................................................................................. 63
Structuur – cellen .............................................................................................................................. 66
Types bindweefsel ............................................................................................................................. 69
Functie ............................................................................................................................................... 71
Regeneratie ....................................................................................................................................... 71
Het kraakbeen ....................................................................................................................................... 71
Kraakbeentypes ................................................................................................................................. 72
Groei .................................................................................................................................................. 74
Voeding en degeneratie .................................................................................................................... 74
Het beenweefsel.................................................................................................................................... 75
Functies ............................................................................................................................................. 75
Basisstructuur beenderen ................................................................................................................. 75
Schedelbeenderen............................................................................................................................. 75
Basisstructuur .................................................................................................................................... 76
Basisstructuur beenweefsel .............................................................................................................. 76
Osteocyten ........................................................................................................................................ 76
Basistructuur compact been ............................................................................................................. 76
Basisstructuur .................................................................................................................................... 77
Organisatie (anatomisch) .................................................................................................................. 77
4
, Basistructuur ..................................................................................................................................... 78
Remodellering en vorming ................................................................................................................ 78
Organisatie (histologisch) .............................................................................................................. 78
Remodellering ............................................................................................................................... 79
Beenvorming: osteogenese ........................................................................................................... 79
Intramembraneuze osteogenesen ................................................................................................ 79
Endochondrale osteogenese ......................................................................................................... 79
Breukheling.................................................................................................................................... 81
Gewrichten ............................................................................................................................................ 81
Synarthrodiale gewrichten ............................................................................................................ 82
Tussenwervelschijven .................................................................................................................... 82
Diarthrodiale gewrichten .............................................................................................................. 82
Gewrichtskraakbeen...................................................................................................................... 82
Diarthrodiale gewrichten .............................................................................................................. 83
Gewrichten .................................................................................................................................... 83
Het bloed ............................................................................................................................................... 84
Inleiding ............................................................................................................................................. 84
Erytrocyten ........................................................................................................................................ 84
Celinhoud ...................................................................................................................................... 84
Vorm .............................................................................................................................................. 84
Groepering (uitstrijkje) .................................................................................................................. 85
Kleur .............................................................................................................................................. 85
Levensduur .................................................................................................................................... 85
Parameters (niet vanbuiten kennen) ............................................................................................ 85
Leukocyten ........................................................................................................................................ 86
Leukocyten – granulocyten ........................................................................................................... 86
Leukocyten – agranulocyten ......................................................................................................... 89
Trombocyten ..................................................................................................................................... 90
Morfologie ..................................................................................................................................... 90
Lichtmicroscopisch ........................................................................................................................ 91
LM en TEM ..................................................................................................................................... 91
SEM ................................................................................................................................................ 91
Functie ........................................................................................................................................... 91
Bloedklonter .................................................................................................................................. 92
Afweer ............................................................................................................................................... 92
Hematopoiese ....................................................................................................................................... 92
5
, Prenataal ........................................................................................................................................... 92
Postnataal .......................................................................................................................................... 92
Hematopoietische stamcellen ........................................................................................................... 93
Erytropoiese ...................................................................................................................................... 94
Granulopoiese ................................................................................................................................... 96
Monocytopoiese................................................................................................................................ 98
Trombopoiese ................................................................................................................................... 98
Lymfopoiese ...................................................................................................................................... 99
Hematopoiese ................................................................................................................................. 100
Het spierweefsel .................................................................................................................................. 100
Inleiding ........................................................................................................................................... 100
Skeletspierweefsel – structuur ........................................................................................................ 101
Skeletspierweefsel – contractie ...................................................................................................... 104
Skeletspierweefsel – classificatie .................................................................................................... 105
Skeletspierweefsel – myogenese .................................................................................................... 105
Skeletspierweefsel – hypertrofie .................................................................................................... 105
Skeletspierweefsel – atrofie ............................................................................................................ 106
Glad spierweefsel – structuur ......................................................................................................... 106
Glad spierweefsel – contractie ........................................................................................................ 107
Glad spierweefsel – classificatie ...................................................................................................... 107
Glad spierweefsel ............................................................................................................................ 108
Hartspierweefsel – structuur........................................................................................................... 108
Hartspierweefsel - contractie .......................................................................................................... 109
Hartspierweefsel ............................................................................................................................. 109
Het zenuwweefsel ............................................................................................................................... 110
Inleiding ........................................................................................................................................... 110
Neuronen......................................................................................................................................... 110
Synapsen.......................................................................................................................................... 113
Neuroglia ......................................................................................................................................... 115
Neuroglia – CZS................................................................................................................................ 115
Neuroglia – PZS................................................................................................................................ 118
Zenuwvezel ...................................................................................................................................... 118
Zenuw .............................................................................................................................................. 121
Ruggenmerg .................................................................................................................................... 122
Regeneratie ..................................................................................................................................... 122
Trofische functie .............................................................................................................................. 123
6
, Algemene inleiding
Inleidende begrippen
Cytologie = “cytos” en “logos” celleer, - Morfologische eenheid: cytomorfologie
de cel - Functionele eenheid: celbiologie
Metabolisme (anabolisme, katabolisme)
Groei
Deling (mitose)
Afsterven (apoptose)
Histologie = “histos” en “logos” - Organisatie cellen tot weefsels (epitheelweefsel, spierweefsel,
weefselleer zenuwweefsel, bindweefsel) algemene histologie
- Organisatie weefsels tot organen, stelsels
(spijsverteringsorganen, ademhalingsorganen…) bijz histologie
Anatomie = “anatomia” ontleedkunde Organisatie van organen of stelsels tot individuen
Embryologie = “embruon” en “logos” Ontstaan en ontwikkeling van organen, stelsels, individuen
Morfologie, fysiologie en biochemie Normale toestand
Histopathologie, anatomopathologie, Pathologische toestand
pathofysiologie
Histologische onderzoekstechnieken
Optische principes
Microscopie – eenheden
- Microscopen
Lichtmicroscoop: lichtbundel, µm
Elektronenmicroscoop: elektronenbundel, nm
- Lengte-eenheid in microscopie
SI-eenheid Symbool en waarde
Micrometer µm = 0.001 mm = 10-6m
Nanometer nm = 0.001 µm = 10-9 m
vb. celmembraan ongeveer 9 nm
1 Angström (atomair niveau) 0.1 nm = 10-10 m
7
,Oplossend vermogen en contrast
Oplossend vermogen (R, resolutie)
- Definitie: omgekeerde afstand tussen 2 punten zodat ze nog als
aparte punten kunnen onderscheiden worden
Hoe kleiner de afstand, hoe groter de resolutie
- R = (K*λ)/NA
NA = n * sinα
- R en vergroting…
Contrast
- Dode cellen of weefsels
- Levende cellen of weefsels
De lichtmicroscoop
Gewone lichtmicroscoop
- Doorvallende lichtbundel
- Dunne coupes (3-8 µm)
- Condensor, objectief, oculair
- R = 0.25 µm
Fasecontrastmicroscoop
- Weefsel wijzigt lichtstraal
Breking: scherpte
Wijziging in golflengte: kleur
Wijziging in amplitude: intensiteit
- Verschillende brekingsindexen: faseverschil
- Optisch systeem
Faseverschil: amplitudeverschil: verschillende lichtintensiteit
Donkerveldmicroscoop
- Licht niet gecondenseerd in het objectief, maar rakelings erbuiten
Preparaat buigt stralen af
Vallen gedeeltelijk in objectief
Omlijning van structuren wordt zichtbaar
Polarisatiemicroscoop
- Polarisator en analysator in lichtbundel
- Polarisator tussen lichtbron en coupe: polariseert lichtbundel in één vlak
- Analysator tussen coupe en oculair: laat licht in één vlak door
- Polarisator en analysator gekruist: geen licht doorgelaten
- Evenwijdige structuren in weefsel zullen licht dubbel breken
Worden zichtbaar (anisotroop)
Fluorescentiemicroscoop
- luminescentie (excitatie, emissie)
fosforescentie
Fuorescentie
- Epifluorescentie
8
, - Primaire fluorescentie (aanwezig in molecule) of autofluorescentie
- Secundaire fluorescentie
Geïnduceerd: specifieke componenten aantonen door specifieke binding
Fluorochromen: fluorescerende eigenschappen
- Confocale microscopie, FRET
De elektronenmicroscoop
Algemeen
- Elektronenbundel (kathode, anode)
- Voltage, hoogvacuüm
- Magnetische velden
- Fluorescerende scherm
Transmissie elektronenmicroscoop (TEM)
- Doorvallende elektronenbundel
- Ultradunne coupes (20-100 nm)
- Contrasteren (uranyl- of loodzouten)
- Sub-cellulair, ultrastructureel, R tot 0.1 nm
Scanning elektronenmicroscoop (SEM)
- Elektronenstraal niet doorheen weefsel maar teruggekaatst
- Oppervlak of breukvlak wordt afgetast
- Teruggekaatste bundel w gedetecteerd door detector: stimuleert kathode die fluorescerend
scherm bestraalt
- Voorbereiding weefsel (dehydrateren, coaten)
Methodologie
Paraffinemethode (LM)
- Monstername
Vlug na de door (autolyse, bederf, diffusie, …)
Grootte
- Fixatie (meestal immersiefixatie)
Algemeenheden
Weefsel bewaren zoals het in vivo gestructureerd is
Chemische reactie
Formaldehyde
Formaline (handelsformol, formol) = 40% formaldehyde-oplossing
4% formaldehyde-oplossing (10% formoloplossing- voor weefselfixatie
Voordelen en nadelen
Picrinezuur, alcohol, trichloorazijnzuur, aceton, kaliumbichromaat, …
- Doorvoeren, inbedden en snijden
Consistentie niet voldoende na fixatie
Infiltratie van weefsel met paraffine via tussenstappen (alcohol, xylol)
Coupes: 4-20 µm
- Kleuren (histochemie, autoradiografie, immunohistochemie)
- Nabehandeling
Ontwateren
9
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