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Transcript of The Mouse Placenta - RTP Rodent Pathologyrtprodentpathology.org/...TheMousePlacenta_handout.pdf ·...
1
The Mouse Placenta
Jerrold M. Ward, DVM, PhD, DACVP, FIATP
Veterinary Pathologist
Today’s Discussion Outline
• Comparative anatomy & histology• Embryonic development of the mouse placenta
(extra‐embryonic tissue)• Normal histology• Histopathology ‐ lesion classification• Developmental abnormalities • Infectious diseases• Toxicologic pathology• Tumors• Resources
Origin of the Terminology “placenta”
Modern Latin from L, literally , a cake from Classical Greek plakounta
Πλακούντας
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What is a Placenta?
John King’s Necropsy Show & Tell ‐ submitted by Dr. M Smith, Cornell U., Sheep, Twin lambs in a single amnion
Classification of The Placenta by Gross AppearanceSatoshi Furukawa et al, Nissan Chemical Industries, J Tox Path 27: 11, 2014
Horses, pigsRuminants
Carnivores Primates, rodents
Rats, micePrimates
Rabbits
Carnivores
Ruminants
Horses, pigs
Classification of The Placenta by HistologyS Furukawa et al, J Tox Path 27: 11, 2014
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Extra‐embryonic Tissues
MRI, A Carretero et al in J Ruberte
E12
FromWard &Devor-Henneman2000
Uterus &Embryo
NecropsySheet
Placenta Dissection & Trimminguse formalin fixation
SC Pang et al. InBN Croy et al.Guide to Investigation of Mouse Pregnancy2014
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Fixation and Trimming of The Placenta
IHC, ISH: paraformaldehyde or formalinImaging: formalin or other fixatives
The intact placenta is fixed in formalin (48 hrs), not Bouin’s
E12.5
The Pregnant Mouse Uterus
E 8.5
E 14.5E12.5
Courtesy of P. Treuting
Mouse Placental DevelopmentJ Rossant and JC Cross, Nature Rev Genetics 2: 502, 2001
Jay Cross, DVM, Calgary
Vet Med
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Placental Development E7‐12
From Ward and Devor-Henneman, 2000
EMBRYO
PLACENTA
Pregnancy Dating in the
Rat Placenta
EP De Rijk et alToxicol Pathol30: 271, 2002
Mouse Placenta Histology Section Orientation
JM Ward (for mouse)
Decidua
DeciduaUterus
Uterus
J Rossant and JC Cross
S Furukawa (for rat)
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Uterus
Decidua
Yolk sac Chorionic plate
Labyrinth
The Mature Mouse Placenta E12.5
endoderm
Embryonal&
Maternal
Maternal
GIANT CELLS
SPONGIOTROPHOBLASTS
LABYRINTH
DECIDUA
CHORIONIC PLATE
Yolk Sac
Labyrinth E10 Labyrinth E11
Embryonic erythroid cells
E
EmbryonicLabyrinthtrophoblasts(LTB)
LTB
Embryonic endothelium LTB
LTB
Maternal rbc
Dam’srbc
Dam’srbc
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Labyrinth E13 Labyrinth E15
M
EmbryonicEndothelium
LTB
E
Maturing embryonal erythroid cells
LTB
Mature erythrocytes of embryo and dam
LTB
LTB
EmbryonicEndothelium
Placental Vascularization – micro‐CTMY Rennie, In The Guide to Investigation of Mouse Pregnancy, 2014
Glycogen Cells ‐ E14a trophoblast cell of unknown origin with unknown functions
PAS
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Placental Aging/Degenerative Changes
Membranes
E12
GibbonAM Cater et al, Placenta 2016 Jan;37:65‐71
E14.5
Decidua
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Immune Functions of The Placenta
The Metrial Gland in The RatC Picut et al. Tox Path 37: 474, 2009
Perforin IHCNK marker
Mouse Granulated Metrial Gland?D. Bulmer et al, Cell Differentiation 20: 77, 1987
E14
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Mouse Placental Gene Expression(Gheorghe et al, J Soc Gynec Invest 13: 256, 2006; Gasperowicz et al, Placenta 29: 651, 2008)
DG Simmons, In Guide to Investigation of Mouse Pregnancy BA Croy et al, Eds
Genes Expressed in Mouse Placenta
Endothelium ‐ MECA‐32, CD31
TB giant cells ‐ ProlactinDecidual cells – MAC2
Spongiotrophoblasts ‐ SOS‐2
SM Isaac et al, 2014
Labyrinth TBs
Labyrinth and Junction TBs
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KL Boyd et al., in Treuting, Dintzis & Montine, 2017
The Primate Placenta
Monkey – Cyno, E120
Courtesy of Mark Cline, WFU
Human
M, maternal bloodIVS, IntervillousSpace
SK, Syncytial knot
V, fetal vessel
The Placenta Is Important For Embryonic Growth
From E9‐12
If it develops abnormality or fails, the embryo will die due to lack of normal nutrition received
from the dam through the placenta
Possible Causes of Embryonic Mortality E9‐12
• Placental failure• Membrane abnormalities
• Cardiovascular abnormalitiesAbnormal embryonal blood vessel
developmentHeart failure
• Abnormal erythropoeisis (in yolk sac/embryo)
• Loss of normal cell cycle regulation
• Cell adhesion/germ layer defects/patterning
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Placental Failure As A Cause of Embryonic Lethality
• Inactivation of a gene important for placental development and function
• Toxic or other changes to specific placental anatomical components
• Functional changes in the placenta that interfere with nutrition and oxygenation of embryo
• Changes that interfere with normal embryo development
• Most placental changes cause embryonic lethality prior to birth
How to Evaluate Placental Causes of Embryo Lethality
• Determine patterns of gene expression in placenta, membranes and embryo at various stages of gestation – ISH/IHC/Northern blotting/other methods
• Determine cells and anatomic placental/embryonal structures normally expressing the gene
• In null mice, these cells may not function normally for placental development
• Null mutation abnormal placenta developmentembryonic death
Jackson LaboratoryMammalian PhenotypeBrowser
http://www.informatics.jax.org/vocab/mp_ontology/MP:0001711
558 genotypes1214 annotations
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Which Comes First?Placental Lesions or Embryonic Lesions
or Are They Concurrent?
Patterns of Lesions in the Placenta
Bolon & Ward2015
Developmental Abnormalities in The Mouse Placenta
• Labyrinth blood vessel formation (94 genotypes) ‐ Vhl, Tfeb, Dlx3, Ppar, Mash2 (Ascl2), Hgf, Egfr, Lifr, Il11ra, Err‐JunB, Hsp90Cyr61, Rap250 (Ncoa6)
• Labyrinth trophoblasts ‐ Vhl, Egrfr, Ascl2, Rxra, Arnt, Lifr, Err‐, l‐mfa (Mdfi), Hsd17b2, Cx26 (Gjb2)
• Spongiotrophoblasts ‐ Arnt• Giant cell trophoblasts ‐Mdfi, Cdkn1c
• Giant placenta – cloned mice, Cdkn1c
• Decidua ‐ Il11ra
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Blood Vessels of the Placental Labyrinth
•Angiogenesis – the formation of new blood vessels from pre‐existing blood vessels (endothelium)
•Vasculogenesis ‐ the formation of new blood vessels when there are no pre‐existing ones; formation of new blood vessels from mesenchyme, in mouse embryo from the chorionic plate (CP)
No Nulls At BirthDNA Analyzed From Yolk Sac
+/+, +/‐, but no ‐/‐ (no yolk sac found grossly or in HE slides)
Vascular Lesions in the PlacentaJR Gnarra et al, Proc Natl Acad Sci U S A 94: 9102, 1997
Dead Embryo
E12.5Lack of normal vasculogenesis
disruption of normal placental development
+/+ Vhl -/-Hemorrhagicplacental sites3/12 were KO
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Laser Capture Microdissection Performed
16 yolk sac laser capture microdissection‐procured DNA analyzed14 were ‐/‐2 were +/‐
Abnormal Vasculogenesis of The LabyrinthCD31 IHC
CD31 (formalin, Santa Cruz M-20, goat anti-mouse,1:500, antigen retrieval)
Labyrinth
Labyrinth
Chorionic Plate
+/+ Labyrinth E 12.5 Vhl ‐/‐
Labyrinth
Trophoblast Degenerative Lesions Eosinophilic Droplets
Dysplasia, HyperplasiaSyncytial Labyrinth Trophoblasts
Necrosis
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Abnormalities of Cell Cycle Regulation
• Increased apoptosis•Cell proliferation (Rb1 in placenta)• Imbalance between apoptosis and cell proliferation
•Brca1, Brca2, Rad51, Braf, Kras, Ski, p130 (Nolc1), DNA ligase IV (Lig4)
•Cell cycle abnormalities were usually reported in the embryo and not placenta
Trophoblast Hyperplasia from Rb loss in Tb stem cells PL Wenzel, A De Bruin, et al, Genes & Development 21: 85, 2007
Placentomegaly of Cloned EmbryosS Tanaka et al, Biol Reprod 65:1813, 2001 ;C. Palmieri et al, Vet Pathol 45: 865, 2008
Expansion of spongiotrophoblasts with increased glycogen cells
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Abnormalities of The Yolk Sac
•Mesodermal defects ‐ alpha‐5‐integrin (Itga5), Fn1
• Endodermal defects – Fn1, Hand1•Vascular hypoplasia ‐ Itga5, MEF2C•Vascular dysgenesis ‐Myc, Hand1, Np1/Np2•Dysgenesis of erythropoiesis ‐ Tf, Scl, Tgf‐1•Hypoplasia of vitelline vessels ‐ Tf, c‐myc, Scl
Yolk Sac lesions
Ballooning degeneration, apoptosis – E16
No vessels or no blood?
Defects inVasculogenesis
&Hematopoiesis
TGF1 -/-
Dic
(Dickson et al 1995)
Defective haematopoiesisand vasculogenesisin transforming growth factor‐beta 1 knock out mice
MC DicksonDevelopment121: 1845, 1995
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Abnormalities of the Chorion & Allantois
• Lack of or abnormal fusion ‐ Vcam1, alpha‐4 integrin (Itga), Rbp‐Jk, Smad1, Lpp (Ppap2b)
• Mesodermal defects ‐Mash2 (Ascl2)
• Allantoic ballooning ‐ alpha‐4 integrin (Itga)
• Chorionic ectoderm defects ‐ Hand1
• Allantoic defects ‐Myc
Absence of Chorioallantoic Fusion
VCAM-1 null, GC Gurtneret al, Genes Dev 9:1, 1995
Infectious Disease
• Bacteria (Brucella, Streptococcus, Listeria, Mycobacterium, Treponema)
• Fungi (Candida)
• Viruses (CMV, Zika, Herpes, HIV, others),
• Protozoa (Trypanosomes, Plasmodium)
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Placental Pathology of the Pregnant Mouse Inoculated with Brucella abortus Strain 2308
L. T OBIAS, et al, Virginia Tech Pathobiology, Vet Pathol 30: 119, 1993.
BALB/c mice, ip at E9, sac at E18, necrosuppurative placentitis
IHC
Massive Decidual Necrosis: Plasmodium chabaudiM Waisberg, M Orandle, et al, PLoS One. 2013; 8(5): e62820 E8
Zika Virus Infection during Pregnancy in Mice Causes Placental Damage and Fetal Demise
JJ Miner et al, Cell 165: 1081, 2016
Normal mice – clear virus
Kos ‐Ifnar1‐/‐ mice(lack of Type I interferon signaling)
develop placental infection(trophoblasts, endothelial cells), and brain infection
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Toxicology ‐ Rat PlacentaS Furukawa et al, Exp Tox Path 67: 443, 2015; 65: 211, 2013
Chlorpromazine‐induced placental toxicity
Cisplatin
Tumors of the Placenta
• Choriocarcinoma
• Yolk sac carcinoma
• Deciduoma
• Granular cell tumor?
BM Bany In The Guide To Invest In Mouse Pregnancy
SummaryHow to Determine if Placental Failure Occurs
• E9.5‐10.5 determine if embryo is grossly normal
• Determine if there are any histopathological embryonic lesions that could cause death
• Check especially placenta, heart, blood vessels, yolk sac, and any tissues where gene is expressed
• Placenta – determine if lesions found occur prior to early lesions of embryonic death
• Embryonic lethality may occur from lesions in both placenta and embryo
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Mouse Placenta References• Comparative Placentation (K. Benirschke)
http://placentation.ucsd.edu/
• Rossant, J & Cross, JC. Placental development: lessons from mouse mutants. Nature ReviewsGenetics 2: 538‐548, 2001
• Ward & Devor‐Henneman, Gestational mortality in genetically engineered mice. In: Ward, Mahler, Maronpot, Sundberg, Pathology of Genetically Engineered Mice, Ames: Iowa: ISU Press, p.103‐122, 2000
• Ward JM, S Elmore & J Foley, Pathology methods for the evaluation of embryonic and perinatal developmental defects and lethality in genetically engineered mice. Vet Pathol, 49: 71, 2012
• Natale DR, Cross JC, et al, Phenotypic analysis of the mouse placenta, Methods Mol Med. 2006: 121:275‐93, 2006
• De Rijk EP, et al, Pregnancy dating in the rat. Toxicol. Pathol. 30:271‐282, 2002
• Furukawa S et al, Toxicological pathology in the rat placenta. J Toxicol Pathol 24:95, 2011.
• K Benirschke, GJ Burton, Pathology of the Human Placenta, Springer, 2012
• JM Cline et al, The Placenta in Toxicology. Part III: Pathologic Assessment of the Placenta, ToxicolPathol 42: 339, 2014
• A Croy, AT Yamada, FJ DeMayo, SL Adamson. The Guide to Investigation of Mouse Pregnancy, Academic Press, 2013
Mouse Embryo References
• ME Dickson et al, High‐throughput discovery of novel developmental phenotypes. Nature 537: 508, 2016
• Theiler K, The House Mouse, http://www.emouseatlas.org/emap/ema/theiler_stages/house_mouse/book.html
• Kaufman, MH. The Atlas of Mouse Development, Academic Press, 1992; Kaufman, MH & Bard JBL, The Anatomical Basis of Mouse Development, Academic Press, 1999; Kaufman MH, Nikitin & Sundberg, Mouse Endocrine System Development, 2010 ‐ http://www.emouseatlas.org/emap/eHistology/
• Ward JM, S Elmore & J Foley, Pathology methods for the evaluation of embryonic and perinatal developmental defects and lethality in genetically engineered mice. Vet Pathol, 49: 71, 2012
• Rossant, J and Tam, PPL, Mouse Development, Academic Press, 2002
• Kaufman, MH, Nikitin & Sundberg, Histologic Basis of Mouse Endocrine System Development, (with digital slides), CRC Press, 2009
• Papaioannou VE & Behringer , RR, Early embryonic lethality in genetically engineered mice: diagnosis and phenotypic analysis. Vet Pathol 49: 64, 2012
• A Croy, AT Yamada, FJ DeMayo, SL Adamson. The Guide to Investigation of Mouse Pregnancy, Academic Press,2013
• Prenatal brain atlas http://www.epmba.org