The Sertoli Cell Junction Dynamics

Falana, Benedict Abiola Department of Anatomy,

CMUL. Matriculation Number: 109091016

ANA 902

25th February 2014

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OUTLINE

• INTRODUCTION• LITERATURE REVIEW• DISCUSSION–Sertoli cell structure–Sertoli cell function–Sertoli cell JUNCTIONS

• CONCLUSION• REFERENCES

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INTRODUCTION: List of Abbreviations

BM= Basement Membrane ECM= Extracellular Matrix BTB= Blood-Testis Barrier TJs= Tight Junctions ES= Ectoplasmic

Specializations NAMPC= Nectin-Afadin

Multi Protein Complex ILMPC=Integrin-Laminin

Multi Protein Complex TT= Testosterone FSH= Follicle Stimulating

Hormone GDNF= Glial cell line

Derived Neutrotrophic Factor

CCMPC= Cadherin-Catenin Multiprotein Complex

IPEC-J2= Intestinal Epithelial cells

CAM= Cell dhesion Molecule

AC= Adenylate Cyclase MDCK= Mardin Darby Canine

Kidney CLASP= Cytoplasmic Linker

Associated Protein PI-3= Phosphatydyl Inosotol PLA2=Phospholipase A2, cAMP=Cyclic Adenosine

Monophosphate RHOB= Ras Homolgue B PKA= Protein Kinase A MAP Kinase= Mitogen Activated

Protein Kinase FGF- Fibroblast Growth Factor ICAM= Intercellular Adhesion

Molecule ST- Seminiferous Tubule FAK- Focal Adhesion kinase

NOS= Nitic Oxide Synthase CATNB= Beta Catenin B GJ= Gap Junction AJ= Adherens Junction AnJ= Anchoring Junctions CAM I = Calmodulin I NR5A1= Nuclear receptor

steroidogenic factor RAI 14- Retinoic acid

inducible protein 14 ZO- Zona Occludens CX43- Connexin 43 SRY- Sperm Region Y

TRAF4=TNF receptor associated factor

TM3= Transmembrane Segment 3 ILK= Integrin Linked Kinase ERM= Ezrin Radixin & Moesin GDNF=Glial Cell Line Derived

Neurotrophic Factor MIS=Mullerian inhibitory Substance JAM= Junction Adhesion Molecule TGFβ= Transforming Growth Factor

Beta ERK= Extracellular signal Regulated

Kinases ECL2= Extracellular loop 2 TNF= Tumor zNecrotic Factor SOX= SRY-related HMG Box GATA=Globin Transcription Factor

Introduction: SPERMATOGENESIS• 1. Spermatogonia– Proliferate by mitotic divisions

to provide stem cells & cells which will proceed through spermatogenesis (1º spermatocytes)

• 2. Spermatocyte– diploid cells (2n) give rise to haploid

cells (1n)

– 1º spermatocytes enter Meiosis I to form 2º spermatocytes which then enter Meiosis II and result in spermatids

• 3. Spermatids– spermatid differentiation into

spermatazoa 5

INTRODUCTION

Enrico Sertoli (University of Pavia Italy 1865)

1862; discovered Sertoli cell

Published in 1865

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Enrico Sertoli (1842 – 1910)

Questions• What is the mechanism of by which

developing germ cell traverse the epithelium?

• What signalling event determine the time of for germ cell to translocate from one site to the other?

• What interactions between Sertoli and germ cells take place during translocation?

• What are the events leading to formation and disruption of Sertoli-germ cell junctions?

Ultrastructure of the Sertoli Cell: Irregular nuceli, Mitochondria, lipid and RER are prominent.Courtesy : www.naturescience.com

• Ultrastructure of the Sertoli Cell: Irregular nuceli, Mitochondria, lipid and RER are prominent

LITERATURE REVIEW

On going Controversies.

• A Single Spermatogoonium (2n) gives rise to Eight Spermatids (1n)

(de Krester & Kerr 1988).

• Preleptotene and Leptotene spermatocytes. (Zipper Theory)

• BTB. (Byers 1993; Pelletier 2001).

• Sertoli-Germ cell bi-directional secretion of Androgen-binding protein. (Gunsalus & Bardin 1980).

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Table 2: Proteins Secreted by Sertoli Cells

• a Table modified from Griswold, 1988.

Hormones or Growth FactorsTGF- precursorAnti-Muillerian hormone (MIS)InhibinTGF-EGF-like growth factorBasic FGF-like growth factorSeminiferous growth factorIGF-1Interleukin-l-like factorLeydig cell stimulatory factor Basement Membrane ComponentsType IV collagenLaminin

EnzymesProcathepsin L (cyclic protein 2)Plasminogen activator Transport proteinsAndrogen-binding proteinTransferrinCeruloplasmin OtherSulfated glycoprotein 1 (prosaposin)Sulfated glycoprotein 2 (clusterin)TestibuminTestins

TABLE 1: Germ Cell-Sertoli Cell

Effects on Germ Cells Effects on Sertoli Cells

Limited differentiation in co-cultures

DNA and RNA synthesis

Protein synthesis

Viability and ATP levels

Adenylate cyclase activity

Protein phosphorylation

Alterations in glycoprotein composition

Alterations in membrane proteins

Transferrin secretion

ABP secretion

Estradiol secretion

Cyclic variations in secretory products

Reglation of Spermatogenesis

FSH.LH.TT.ABP.

(McLachlan et al., 2002 ; Kerr et al. 2006; ).

SRYSOX HMGFGF GATAFOGGADD 45gNRSA1 (Hiramatsu 2010; Pask 2010; Bormann 2011;

Johnen 2013)

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• CX43 alters Occludin expression in the rat ST.

(Gerber 2014)

• Spectrin and Plectin sorrounds the actin cuffs of apical tubulolobular complexes in the rat.

(Aristaeuss de Asis 2013)

• Clathrin/actin-base endocytic machinery is associated with junction turnover in ST.

(Vogl 2014).

• Dicer is required for sertoli cell function and survival.

(Papaioannou 2009; Kim 2010;Hensley et al., 2014).

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• RIA 14 regulates F-actin dynamics at the ES.

(Qian 2013).

• CAM-I enhances Germ-Sertoli cell interaction in spermatogenesis.

(Lewis 2005; Wakayama & Iseki 2009)

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• FAK is a regulator of F-actin dynamics (Li 2013)

• Disruption of Sertoli-germ cell adhesion function limited to adherens junctions.

(Xia et al., 2005)

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• Taurine disrupts the morphology and Ultrastructure in the testis of mice.

(Abdel-Moneim 2013)

• TGF-beta3 regulates anchoring junction dynamics via the Ras/ERK signaling pathway.

(Xia 2005)

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• Ultrastructural changes of the Sertoli and Leydig Cells following STZ induced diabetes.

~ Body Weight ↓ (P˂0.05)

~ Testicular Weight↓(P˂0.05) in diabetic rats compared with controls.

(Kianifard 2012)

• Β- Conglycinin reduces the tight junction occludin and ZO-I expression in IPEC-J2.

(Brosnan 2012 ; Zhao et al., 2014)

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• Palmitoy-protein thioesterase I

(PPTI) an obesity induced testicular

marker of reduced fertility. (Liu 2014)

• Claudin-3 and Claudin-5 folding and assembly into tight junctions are controlled by non- conserved residues in TM3 and ECL2 segments. (Rossa et al., 2014; Shabazi 2014).

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• Immunohistochemical analysis of Histone H3 modifications in germ cells during mouse spermatogenesis.

(Song et al., 2011).

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• TRAF4 impedes the formation of tight junction integrin-linked kinase (ILK) Talin-1.

(Rosseau et al., 2014).

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DISCUSSION

STRUCTURE Blood-testis barrierControls the entry and exit of

nutrients hormones and other chemicals into the tubules

Makes the ad luminal part of ST, an immune privileged site.

(Wakayama 2009; Kopera 2010)

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SECRETION:

MIS

Inhibin

Activin

ABP

Estradiol aromatase. 27

GDNFERMTransferrin

(Xiong et al., 2006)

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Hormones,Growth factors,Proteases (Catepsins,Metzincin,Plasminogens) Protease inhibitorsComponents of the extracellular matrix

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Junctions

The BTB

Tight junctions(TJs) Ectoplasmic specializations (BES)Tubulolobular complex (BTC) Desmosome-like junctions Hemidesmosomes

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Blood Testis Barrier- endocrine reviews

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BTB

FUNCTIONSAllow sertoli cells to control the

ad- luminal compartment.Regulates the chemical

composition of luminal fluid.Sensitive to trauma and Autoimmune response

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TJs

COMPOSITION:

Fused membranesBead-like ComponentsSpan adjacent membraneFormed by strands of

transmembrane proteins .

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TJs

FUNCTIONS

Barrier FunctionImpermeableRegulates absorptionCreates a seal

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AJs

• Zona adherens

• Located at site of cell-cell interaction

• Actin-linked

• Attaches ECM

• Posses specific transmembrane receptors of the integrin family

• Adhesive function

Cadherin

Ca ion dependent CAMadhesion at adherens junction

Role:Teetheres cells to the ECM.Tranduce signals.Proliferation. Migration . Differentiation .

AnJ

• Desmosome-like junctions

• Ectoplasmic specializations

Key• 1.Peritubular cells

2. Basal membrane3.Spermatogonia4. Tight junction5. Spermatocyte16. Spermatocyte27a. spermatid7b. Spermatid8. Acrosome9. residual bodies10.Sperm cells11. Nucleus of sertoli cellA. Basal zoneB. Adlumunal zone

• www.instantanatomy.com.39

Desmosome-like Junctions

COMPOSITION

Macula adherensFound between Sertoli cells at the

BTBGerm cells up to but not including

step 8 spermatids. (Holthofer et al.,2007).

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Desmosome-like junctions

• COMPOSITION

Desmoglea. Dense Cytoplasmic plaques.

(Garrod et al., 2005; Holthofer et al., 2007; Scothern & Garrod 2008)

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Desmosome-like junctions

COMPOSITION: Cadherin family (a) Desmogleins (Dsg) (b) Desmocolins (Dsc) © IntegrinsAssociated with keratin & VinculinDistinct at molecular levelCalcium ion dependent (CAM)Sites of signal transduction

(Bruce et al., 2000)

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Desmosome-like Junctions

• FUNCTIONS~Mediate cell adhesion ( Delva et al.. 2007)

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Desmosome-like junctions

FUNCTIONS: Cell adhesion Cell proliferation, Cell differentiation, Cell migration and Morphogenesis.

(Garrod & Chidley 2004)

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ES

CCMPC

CATENIN ( α-β-ϒ-) :Cytoplasmic proteins Bind to the C-terminus of the type I/II and

desmosomal cadherins Regulates cadherin-mediated adhesion via

the actin and intermediate filament cytoskeletons

( Das et al., 2014)

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ES

CCMPC (Wine & Capin 1999;Johnson & Boekelheide 2002; Lee et al., 2003, 2004; Yan et

al., 2008a; Delva & Kowalczyk 2009; Yan et al., ; 2008b; Izumi et al., 2006).

• Best studied actin-based adhesion unit.

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Regulates:Cytoskeleton Cell polarity,Control of cell division andTumor suppression

• ( 5 distinct subfamilies, type 1 and 2, desmosomal, atypical, and cadherin-like)

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ES

NAMPC

Nectins (1-5) Nectin-like molecules ( Necls,

Necls1-5)

--Ca++-independent

--immunoglobulin-like moles.

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ES

Role:Adhesion ProliferationDifferentiationSurvivalMigrationCell polarity.( Takai & Natkanishi 2003; Takai et al., 2003, 2008c; Irie et al., 2004)

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ES

ILMPC

Functions

• Connects a cell to ECM (Margadant et al., 2008; Geiger et al., 2009)

• Constituents of the BM (Miner & Yurchenco 2004; Miner 2008)

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SUMMARY

• The integrity of the BTB cannot be compromised.

• TJ on and off switch

• Signal transduction is very essential and crucial in tight junctional dynamics.

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CONCLUSION

• Changes in secretory activity of either Sertoli or germ cells initially take place at the level of cell junctions.

• Biology of desmosome-like junctions and ES needs to be thoroughly investigated.

• Basis of male contraception and infertility treatment.

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Acknowledgement

• Sincere gratitude to my able supervisors

Dr. Francis Duru and Dr. Abraham Osinubi for their mentorship role.

• I also want to appreciate my Teachers Dr. Ibeabuchi, Dr. Oremosu, Dr. Kusemiju, Dr. Dosumu, Dr. Olabiyi, Dr. Gbotolorun , Dr. Yama for their assistance during preparation for the review.

• A big thanks to fellow Postgraduate students and the non-teaching staff of the Department.

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