Role of Seminal plasma proteins in semen by J S Rajoriya.ppt

Role of Seminal plasma proteins in semen

2. Semen–A thick ,whitish liquid containing sperm that is produced
by the sex organs of men and male animals
The ejaculated semen consists of two major components viz.
sperm cells (spermatozoa) and the fluid part obtained after
centrifugation called seminal plasma
or
Semen is a complex mixture consisting of spermatozoa and the
liquid known as seminal plasma
INTRODUCTION

4. SEMINAL
PLASMA
(CONSTITUENTS)
IONS
ENERGY
SUBSTRATE
(FRUCTOSE)
BULL
ORGANIC
COMPOUNDS
PEPTIDES
PROTEINS

5. SPERM MOTILITY
CAPACITATION
ACROSOME REACTION
FERTILIZATION
CELL PROTECTION
GAMETE FUSION
Proteins are the main macromolecules contained in seminal plasma,
which have been correlated with male fertility in different species
FUNCTION OF PROTEIN

6. Seminal plasma proteins
•Partly originates from the blood plasma by exudation through the lumen of
the male genital tract and partly are synthesized and secreted by various
reproductive organs
•These proteins are involved in regulation of osmotic pressure and pH of
seminal plasma, transport of ions, lipid and hormones. The biosynthesis and
secretion of these proteins is regulated by testosterone levels in the blood
PRE ALBUMIN
ALBUMIN
GLOBULIN
TRANSFERRING ,
(ALPHA –ANTI
TRYPSIN)
BETA-GLYCOPROTEIN
,( BETA –LIPOPROTEIN)
IMMUNOGLOBINS
,( PEPTIDES HORMONES )
ORSOMUCOID,( KININOGEN

7. •Identified proteins
Forward motility proteins
IgG–Fc binding proteins
Immobilin
Cellular retinol binding
proteins
Andogen binding
proteins
Seminal plasma
Interleukins
Ferrisplasn
Alpha lactalbumin
Clustrin
Heparin binding proteins
Osteopontin
Calsemin
Insulin like growth factors
BSA-A1, BSA-A2
gossact
BSA-A3, BSA-30kDA
proteins
BSP proteins

10. Bovine seminal plasma contains four similar acidic proteins,
previously designated as BSP (bovine seminal plasma)-A1, BSP-A2,
BSP-A3, and BSP-30-kDa, that when added to pituitary cell cultures
result in the immediate secretion of gonadotropins (follitropin and
lutropin)
The amino acid composition is similar to apoA-I
The binding factor cross-reacts in the radioimmunoassay of apoA-
I
The purified binding factor identified as apolipoprotein A-I (apoA-I)
Apo lipoprotein A-I

11. The BSP protein signature is characterized by two tandemly
repeated fibronectin type 2 (Fn2) domains.
Analysis of the relationships between all Fn2 domain-
containing proteins revealed that the Fn2 domains found in
BSP-related proteins have special features that distinguish
them from non-BSP-related proteins.
These features can be used to identify new BSP protein-
related sequences.
Fibronectin type 2

12. In cattle (Bos taurus), the reservoir is evidently formed when
sperm bind to fucosylated ligands resembling Le(a)
trisaccharide on the surface of oviductal epithelium
This protein inhibited binding of sperm to oviductal explants
Blots of SDS-PAGE gels of seminal plasma proteins
revealed the presence of a Le(a)-binding protein with an
apparent mass of 16.5 kDA
Fucose-binding protein

13. Insulin-like growth factor
Insulin-like growth factor-I and -II (IGF-I and IGF-II) are
ubiquitous peptide hormones that exert potent mitogenic,
metabolic, and differentiating actions on cells throughout the
body.
IGFs are produced in the liver and several other organs, and
are frequently released in response to pulsatile secretion of
growth hormone from the anterior pituitary.
In the male reproductive tract, IGF-I has been identified in the
testis, where it is secreted by Leydigand Sertolicells.
IGF-I is believed to be involved in spermatogenesis and
steroidogenesis.

14. The structure of the BSPs show two type II domains
or (fibronectin gelatine binding domains) arranged in tandem
fashion.
Heparin-affinity chromatography was then used for the
separation of GSP-20 and -22 kDa from GSP-14 and -15 kDa.
Amino acid sequence analysis of these proteins indicated
that they are homologous to BSP proteins.
Gelatine-agarose adsorbed Protein

15. A basic protein of apparent molecular weight 15 kD,
designated bSVSP15, was purified from bovine seminal
vesicle secretion
When added to cAMP phosphodiesterase , bSVSP15
inhibited the activation of enzymatic activity by calmodulin in
a reversible manner
Calmodulin has been reported to stimulate various calcium-
dependent enzymatic activities through specific associations
with calmodulin-binding proteins
Calmodulin-Binding Protein

16. Major proteins of mammalian seminal plasma are members of two
main families of proteins namely spermadhesins, which are the major
proteins of boar and stallion seminal plasma and those proteins that
contain fibronectin type II domains, which are the major proteins of bull
seminal plasma
Sperm adhesins are secretory products of the seminal vesicles with the
exception of AWN-1which is present on epididymal sperm and also
synthesized by the rete testis and tubuli recti
Porcine and equine sperm adhesins display carbohydrate-binding
activity
Spermadhesins

17. BSP proteins interact with heparin, apolipoprotein A-I
(apoA-I) and apoA-l associated with high-density
lipoprotein (HDL)
The HDL and heparin present in the female reproductive
tract have been implicated in sperm capacitation and the
acrosome reaction (AR)
BSP proteins modulate the process of capacitation
induced by heparin, HDL, and apoA-l-liposomes
High-Density Lipoprotein

18. Cysteine-rich SecretoryProteins
The cysteine-rich secretory proteins (CRISPS)
represent an evolutionarily highly conserved family
originally described in the rodent male reproductive
tract and more recently in the salivary glands
In the mouse, CRISP-1 is mainly found in the cauda
epididymis where it is synthesized by the principal
cells and secreted into the lumen, accounting for
about 15% of the protein content of the epididymal
fluid

19. Ubiquitin was discovered as a protein capable of inducing B-
lymphocyte differentiation in 1975
Ubiquitin was found in all eukaryotic cells playing a central
role in intracellular proteolysis
Bovine ubiquitin standards were between 0. 16 and 2.5 μg/ml.
Ubiquitin

20. Macrophage migration inhibitory factor
Macrophage migration inhibitory factor (MIF) was
first described as a T-cell cytokine. Since its
discovery as a pro-inflammatory mediator
When secreted by Leydig cells, MIF modulates
Sertoli cell inhibin production
MIF is one of the proteins transferred to spermatozoa
during the epididymal transit

21. Cellular Retinol binding Protein (CRBP)
Retinol (vitamin A alcohol) is essential for male reproductive
function.
Adluminal germ cells also contain two vitamin A-binding
proteins: cellular retinoic acid-binding protein (CRABP) and, in
lesser amounts, cellular retinol binding protein (CRBP)
CRBP is present in relatively high levels in Sertoli cells, where its
level of expression varies dramatically with the spermatogenic
cycle.

22. IgG –Fc binding proteins
IgG-Fc binding protein has been reported in human seminal plasma,
which specifically binds with the Fc region
This protein is synthesised by prostate and is sanative to pronase, but is
resistant to glycosidase and deoxyribonuclease
IgG-Fc binding protein is adsorbed to the sperm surface after
ejaculation and is responsible for the biding of IgG-Fc molecule to
spermatozoa

23. Immobilin is a high mol.wt. (400 kDa) glycoprotein which leepsthe rat
caudaepididymalspermatozoa in fully immobilized state
Immobilin is synthesised by the principle cells of the caput epididymis
and is secreted in the lumen of the tubule which travels with the sperm in
caudaepididymis
Immobilin inhibits sperm motility in the rat and hamster cauda
epididymis mechanically by creating a highly viscous-elastic
environment in the caudaepididymis, which is the store house of
spermatozoa
Immobilin

24. Osteopontin (OPN) is a secreted extracellular matrix phospho
protein identified in various tissues and fluids including those
of the male and female reproductive tracts
OPN was previously identified as a 55 kDa high fertility
marker in bull seminal plasma, produced by the ampulla and
the vesicular gland
When secreted into the proximal tubule of the mouse
nephron , OPN suppressed the accumulation of calcium
oxalate crystals
Osteopontin (OPN)

25. Forward Motility Protein (FMP)
Sperm motility is a key factor in allowing us to determine
semen quality and fertilizing capacity
cAMP activates sperm cytosolic cAMP dependent protein
kinases, which in turn phosphorylate multiple intra-sperm
phosphoproteins that may regulate flagellar motility
These proteins could also play a vital role in diagnosis and
treatment of male infertility, development of contraceptive
vaccines, and population control

26. Androgen-binding protein (ABP) is a glycoprotein (beta-
globulin) and 70-90 kDa produced by the Sertoli cells in the
seminiferous tubules of the testis that binds specifically to
testosterone (T), dihydrotestosterone (DHT), and 17-beta-
estradiol
ABP’s production is regulated under influence of FSH on
Sertoli cell, enhanced by insulin, retinol, and testosterone
Heterogeneity of ABP could be attributed to variation in
glycosylation, phosphorylation , methylation , sulphation and
acetylation pattern of ABP molecules
Androgen-binding protein

27. Heparin-binding Proteins
Heparin binding proteins are abundant on the surface of
ejaculated spermatozoa but are less abundant on the plasma
membrane of epididymal spermatozoa
Heparin-binding proteins also allow spermatozoa to respond
to a challenge with lysophosphatidyl choline and undergo the
acrosome reaction
Heparin-binding proteins (HBP) present in bull seminal fluid
bind to cauda epididymal sperm in vitro and increase the
frequency of zonae pellucidae induced acrosome reactions by
epididymal sperm incubated with heparin

28. Non-heparin binding protein
Seminal plasma proteins were separated on their heparin
binding properties as heparin binding (HBP) and non-heparin
binding (NHBP)
NHBP was found to provide some degree of protection to buffalo
spermatozoa against cryopreservation stress as well as the
deleterious effect of HBP during cryopreservation

29. Clusterin
Clusterin, has recently been identified in the blood, seminal
plasma, and several solid tissues of many species
Clusterin was first identified in the male reproductive tracts of
the rat and ram, where it was named sulphated glycoprotein-2
(SGP-2) and clusterin, respectively
Semen samples with many clusterin-positive spermatozoa
have lower fertility, as determined by non return-to-estrus rates
and had many morphological abnormalities
The presence of clusterin on ejaculated sperm may indicate
improper spermatogenesis or irregular epididymal maturation

30. Seminal plasmin
A highly potent antimicrobial , transcript inhibitory, 5.4 kDa, 48 amino
acids, basic protein present in the bull seminal plasma
Seminal plasminis synthesised and secreted by seminal vesicles,
prostate and ampullaeof the vas deferens of the bulls not by the testis
and epididymis
Seminal plasminis a potent antimicrobial protein present in the bull
seminal plasma and seminal vesicle fluid acting on the wide variety of
gram positive and gram negative bacteria

31. A heat stable, leucine rich seminal plasma protein of 16 kDa
has been isolated and purified from human seminal plasma
and is designed as Gossact
Gossypol inhibits lactate dehydrogenase (LDH) in human
spermatozoa viz. mitochondrial damage, and acrosomal
fragmentation leading to infertility
The inhibitory effect of gossypol of LDH is neutralized by
gossact
Gossact

32. Extracellular matrix proteins have been observed in human
sperm, including fibronectin, laminin, and vitronectin
Vitronectinappears to be localized in the acrosomeof
acrosome-intact sperm and released during the acrosome
reaction
Lamininhas been reported to be localized on the sperm tail
or on the acrosomalmembranes
Extracellular matrix proteins

33. DE, a 37 kD glycoprotein, is a sperm-associated protein that is
synthesized and secreted from the epididymal epithelium in an
androgen-dependent manner and is added the surface of sperm
head during epididymal transit. This protein is also known as
CRISP-1
DE is one of the few reagents that affect sperm-egg fusion
specifically. The human DE-like cDNA is known as AEG-
related protein (ARP) and shares 40% homology with DE
DE (CRISP-1, ARP)

34. Inhibin
Inhibinis a steroid free glycoprotein hormone of 15-30 kDa, present in
seminal plasma and ovarian follicular fluid of various mammalian
species
Inhibinis mainly synthesised by sertolicells in the male and granulosa
cells in female
Inhibinpreferentlysuppresses the secretion and modulates reproductive
functions in the male and female
Physiological effect at pituitary levels by decreasing the basal FSH
secretion