genomic insight of__sperm_motility

Genomic insight of sperm motility

Subodh KumarSenior Scientist

Division of Animal GeneticsIndian Veterinary Research Institute

Izatnagar (Bareilly) UP

1st Feb 2013

Introduction

Apr 12, 2023

2003 2007

Cattle 185.181 199.075

Buffalo 97.922 105.343Total Bovine 283.446 304.765BAHS,

2010

Livestock Population in Million

Cattle Population

DAHD 2007

1997 (Million)

2003 (Million)

2007 (Million)

Growth% (03-07)

Cross Bred 20.09 24.68 33.06 33.92

Indigenous 178.78 160.04 166.6 3.43

10.1%

89.9%

13.1%

86.4%

15.7%

83.6%

Apr 12, 2023

Milk Production (Million Tonnes)

BAHS, 2010

Apr 12, 2023

Milk Production (MT)

BAHS, 2010

% of Cattle contribution

CB 46.98 IND

53.02

IND 81

CB 19

Indigenous Cross Bred

Population (Million) 52.13 12.5

Milk production (MT) 25.357 22.468

Crossbred Contribution

% of Milch cattle Population

Apr 12, 2023

Quality of crossbred semen as compared to exotic and zebu counterparts in the same environment

Parameter B. indicus

B. taurus

B.Indicus x

B. taurus

References

Initial motility(%) 61.00 59.00

79.41

82.19

51.00

67.49

68.13

Singh and Pangawkar (1990)

Shrivastava and Kumar (2006)

Chacon et al.,(1999)

Head + mid piece

Abnormalities (%)

9.4% 15.1 19.0 Chacon et al.,(1999)

Tail Abnormalities +

cytoplasmic droplets(%)

14.0 13.7 18.0

Bull disqualifying rate(%) 54

48

Tyagi et al., (2006)

Chacon et al.,1999

Mass activity 4.00 3.06 Shrivastava and Kumar (2006)

SPD-CMPT (mm) 45.06 39.94

HOST(%) 49.38 42.06

Post Thaw Motility (%) 40.31 28.13 Apr 12, 2023

Higher diseases incidence

Requires intense managemental conditions

Non-availability of superior crossbred germ plasms

(Mathew et al., 1982)

Poor quality semen

(Rao and Rao, 1991)

Poor freezability of semen and cryo-injuries

(Ghosh et al., 2007)

Poor motility and viability of sperms

(Dhanju et al., 2006)

High percentage of dead and abnormal sperms

(Ghosh et al., 2007)

Problems in crossbreds

Apr 12, 2023

Introduction

Sperm motility - a complex process

Parameters of semen quality

Root cause of asthenzoospermia – poorly understood

Understanding of genetic mechanism

Most knowledge based on human and laboratory animals

Sperm motility genes are highly conserved

Sperm Motility - Classification

Morphological basis:* Non-motile sperm * Motile sperm

@ Progressively motile sperm @ Non-progressively motile sperms

* Total motility

Physiological basis:* Activated motility * Hyperactivated motility

Freshly ejaculated sperm

Flagellum generates a symmetrical lower amplitude wave

Move in a straight line in non viscous media - seminal plasma or semen extender

Activated motility

Hyperactivated motility

Present in sperm after reaching Oviduct of female reproductive tract

Flagellar beats asymmetric and of higher amplitude

Move in circular or figure 8 trajectory

Helps to penetrate egg vestment

Seen in association with onset of capacitation but are not dependent on each other

Basic Sperm Structure

FLAGELLA

MITOCHONDRIA

ION CHANNELS

METABOLISM

AKAP4

CatSper I

SPAG6

SPAG11

CatSper II

CatSper IIICatSper IV

GAPDS

SMCP

Candidate genes for Sperm Motility

Encodes a member of the AKAP family.

Alternative splicing of this gene results in two transcript

variants encoding different isoforms.

Nearly half of the protein in fibrous sheaths isolated

from mouse sperm is AKAP4.

This protein and two others, AKAP3 and AKAP-80, have

anchoring sites for cAMP-dependent protein kinase and

helps the cAMP/PKA signaling pathway

Targeted disruption- causes defects in sperm flagellum

and motility.

AKAP4 (A Kinase Anchor Protein 4)

Map Location

Human Xp11.2Mouse:XA1.6

Start : 49,842,146 bp End : 49,852,404 bpSize : 10,259 basesORF : 2565bpExons : 6Translation : 854 amino acid

AKAP4 – Map location in Human

AKAP4 location in Mouse

Organization of AKAP4

I II III IV V

156 2133 102 51 96 27

5’UTR 3’UTR

EXONS (6) 2565 bp

INTRONS

PROMOTER

Organism HumanSimilarity

Dog(Canis familiaris)

88.44(n)84.02(a)

Chimpanzee(Pan troglodytes)

98.84(n)98.4(a)

Cow(Bos taurus)

83.87(n)78.7(a)

Rat(Rattus norvegicus)

82.84(n)78.65(a)

Mouse(Mus musculus)

82.98(n)79.27(a)

Orthologos for AKAP4 Gene

Expression pattern of AKAP4 Gene

SPAG6 (Sperm Associated Antigen6)

Encodes an axonemal protein containing eight armadillo repeats

Expressed in Male reproductive tissues particularly Epididymis and Testis tissues

SPAG6-deficient males were infertile. motility defects, morphologically abnormal (loss of the sperm head) disorganization of flagellar structures,

Important for the maintenance of the axonemal central apparatus and structural integrity of mature sperm.

Essential for sperm flagellar motility

Start : 22,674,406End : 22,746,545Genomic Size : 72,141ORF : 1524 bpExons : 11Aminoacids : 507

MAP LOCATION

Human 10p12.2Mouse 16A3

Cattle 13

SPAG6 Gene summary

Organization of SPAG6

II III IV V VI VII IXVIII X

205 191183 173166 152 14524 95 69116

5’UTR

3’UTR

EXONS (11) 1524 bp

INTRONSPROMOTER

Orthologs for SPAG6 gene

OrganismHuman

Similarity


92.93(n)96.27(a)

Cow(Bos taurus)

91.94(n)96.27(a)


87.42(n)95.85(a)

Mouse(Mus musculus)

87.57(n)96.25(a)

Chicken(Gallus gallus)

82.25(n)85.8(a)

SPAG6 Expression

Encodes androgen-dependent, epididymis -specific secretory proteins. The specific functions not been determined.

Single gene derived from two ancestrally independent β-defensin genes joined by read-through transcription

Some isoforms contain regions similar to beta-defensins.

Rat epididymal cells or human colonic epithelial cells transfected with rat SPAG11 could induce sperm motility in immotile immature sperm. ( Zhou et al. 2004)

Important for the acquisition of sperm motility and the initiation of sperm maturation.

SPAG 11 (Sperm Associated Antigen 11)

SPAG11- gene summary

Start : 7,292,686 bp from pterEnd : 7,308,602 bp from pterSize : 15,917 bases Exons : 8ORF : 327 nt Translation : 108 aa

Map location

Human : 8p23.1Cattle : 27q1.2Mouse : 8A1.1

Genomic organization of SPAG11

P1

A A A

98 153 151 107 76 259 104 279

PROMOTER

3’UTR

A- Poly adenylation sites

5’ UTRINTRONS

EXONS

I II III IV V VI VII VIII

OrganismHuman

Similarity


71.17(n)62.16(a)

Mouse(Mus musculus)

72.07(n)61.26(a)

Orthology of SPAG11

Encodes a protein belongs to GAPDS family of enzymes that play an important role in carbohydrate metabolism.

Functions in a NAD-dependent manner to remove hydrogen and add phosphate to glyceraldehyde 3-phosphate to form 1,3-diphosphoglycerate.

During spermiogenesis, play an important role in regulating the switch between different energy-producing pathways, and it is required for sperm motility and male fertility

GAPDS (Glyceraldehyde 3, phosphate Dehydrogenase Sperm specific

GAPDS gene summary

Start : 40,716,154 bp from pterEnd : 40,728,061 bp from pterSize : 11,908 basesExons : 11ORF : 1227 ntTranslation : 408 amino acid

MAP LOCATION

Human : 19q13.1Mouse : 7B1Cattle : 18

Organization of GAPDS

I II VIII IXIII IV V VI VII X XI

67 178 97 107 91 119 82 152 163 98 73

5’UTR 3’UTR

EXONS (1227 bp)

INTRONS

Orthologs for GAPDS gene

OrganismHuman

Similarity


84.49(n)86.07(a)


99.18(n)98.77(a)

Cow(Bos taurus)

85.57(n)86.84(a)


79.49(n)83.33(a)

Mouse(Mus musculus)

79.41(n)82.6(a)

GAPDS expression

GAPDS Protein

68% identical with somatic cell GAPD. GAPDs has a 72-amino acid proline-rich segment at the amino terminal

end that is not present in somatic cell GAPD.

Exists in sperms as the tetrameric molecule bound to the fibrous sheath of the flagellum

Cysteine residues (C21, C94, and C150) are specific for the sperm isoenzyme

Residue C21 involved in the formation of the disulfide bond between the N-terminal domain of GAPDs and fibrous sheath proteins.

Localised in the principal piece of the flagellum

Encoded protein localised in Cytoplasm, Mitochondrion membrane.

Becomes associated with the mitochondrial outer membranes at spermatogenesis

Function: Organization and stabilization of the helical structure

of the sperm & mitochondrial sheath Absence is associated with male infertility, Reduced sperm motility in female reproductive tract Inability to penetrate the oocyte zona pellucida

SMCP (Mitochondria Associated Cystein Sperm rich Protein)

SMCP Gene summary

MAP LOCATIONHuman : 1q21Mouse : 3F1Cattle : 3

Start : 151,117,422 bp from pter

End : 151,124,147 bp from pter

Size : 6,726 basesExon : One ORF : 351 bp length Translation : 116 amino acid

SMCP

The 5’ & 3’ UTR are more conserved (71%) than the coding sequences (59%).

The open reading frame encodes a 116-amino acid protein and lacks the UGA codons.

The MCSP gene in human, baboon, and bovine is more conserved than its counterparts in mouse and rat.

Expression is restricted to haploid spermatids in humans (Aho et al. 1996).

The 5’ UTR of mouse, rat, and human SMCP also has a predicted

stem loop that is involved in regulation of SMCP expression. (Hawthorne et al. 2006).

SMCP expression

Discovery of a New Protein/Gene

• A new ion channel protein discovered (Clapham 2001).

• This was found to be expressed in testis (Ren et al., 2001).

• Expressed on the principal piece of sperm tail. • Normal sperm (having this protein) showed faster progressive

movements.

• Those lacking it (gene disrupted), swim with 1/3rd speed and move more randomly.

(Clapham and Garbers, 2005)

More about the New Protein/Gene

• Mutants were 100% ineffective at impregnating females though they displayed normal mating behavior.

• CASA = mutant sperm’s main motility parameters of path velocity, progressive velocity and track speed were impaired significantly.

• Most notable, mutant sperm lacked the vigourous beating and bending of the tail region.

• Thus disrupting this gene resulted in marked reduction in sperm motility (Clapham and Garbers, 2005)

This gene/protein was called as

(Cation channel of Sperm)

CatSper is an Voltage gated Calcium selective ion channel protein that plays a central role in sperm motility.

This newly discovered protein ‘controls flow of Calcium ions in to tail of sperm’ which cause fibre like contractile proteins (motor proteins) to

produce forceful lashings of sperm tail.

Other researches on CatSper

• In human, subfertile men with defficient sperm cell motility had significantly reduced (3.5 times) expression of CatSper (Nikpoor et al., 2004).

• Sperm with Null CatSper (CatSper -/-) were not able ascend the oviduct

whereas sperm from heterozygote (CatSper +/-) were able to (Suarez et al., 2005).

• Tiny electric currents generated by Ca+2 moving into sperm have also been recorded which were called as CatSper dependent current (iCatSper) and shown to be an alkaline potentiated, voltage activated, calcium selective channel (Kirichok et al., 2006).

• CatSper are highly specialized flageller protein and could be expressed in HEK 293 cell line (Quill et al., 2007).

CatSper Characterization

• The mouse CatSper gene is predicated to have a primary structures of 686 amino acids with six putative transmembrane domains (S1-S6) and the pore region (P) which exists between S5 and S6 (Jin et al., 2005).

• Out of six transmembrane domains, the fourth is a voltage sensor (Jhang et al., 2006).

• Alternative forms of CatSper and conserved pore region (T X D

x W) motif reported in human (Asadi et al., 2006).

CatSper channel

5 different genes encoding CatSper 1,2,3 ,4 & β

Expressed in the plasma membrane of principal piece of sperm tail

Catsper protein is a single 6 TM spanning repeat – closest to Cav channels

S4 segment acts as a voltage sensor, abundance of histidine and arginine residues

Heterotetramerization of CatSper channel

CatSpers 1,2,3 & 4 may interact directly or indirectly and form a functional tetramer

CatSper1 and CatSper2 can associate with and modulate the function of the Ca(v)3.3 channel, which might be important in the regulation of sperm function.

Controls calcium entry to mediate the hyperactivated motility,

CatSper3 - has role in acrosome reaction and male fertility

Catsper3 and Catsper4 knockout male mice were completely infertile due to a quick loss of motility and a lack of hyperactivated motility under capacitating conditions

CatSper is therefore implicated as a potential target to explore the molecular mechanisms of male infertility.

CatSper genes -Function

MAP Location Mouse: 19 AHuman: 11q13.1Cattle: 29

CatSper 1 gene summary

Start : 65,540,799 bp from pterEnd : 65,550,564 bp from pterSize : 9,766 basesORF : 2343 NucleotideProtein : 780 aminoacid

CatSper 1 gene organisation

VIIV XIII XIX XII

27 115 76 61 73 64 144 92 148 114 213 1216

5’UTR 3’UTREXONS (12) 2443 bp

INTRONS

Orthologs for CatSper1 gene

OrganismHuman

Similarity


82.98(n)77.67(a)

Cow(Bos taurus)

75.06(n)62.88(a)


63.84(n)53.94(a)

Mouse(Mus musculus)

67.48(n)57.67(a)

CatSper2 gene summary

Start : 41,707,993 bp from pterEnd : 41,728,338 bp from pterSize : 20,346 basesExons : 12ORF : 1593 nt Translation : 530 aa

Map location

Human : 15p15.3Cattle : 21Mouse : 2E5

CatSper2 gene organisation

I II V VII XIIII VI VIII XIIXIX

32 165 218 57 100 179 125 156 173 69 174 145

5’UTR 3’ UTR

EXONS (12) 1593 ntINTRONS


OrganismHuman

Similarity


84.91(n)79.36(a)


99.37(n)98.86(a)

Cow(Bos taurus)

82.32(n)76.52(a)


74.86(n)70.06(a)

Mouse(Mus musculus)

74.49(n)69.47(a)


Start : 134,331,495 bp from pter

End : 134,375,291 bp from pter

Size : 43,797 bases

ORF : 1197 nt

Translation : 398 aa

Map location

Human : 5q31.1Mouse : 13B2


VIIIII VII VIII IV VII

5’UTR3’UTR

98 154 240 183 141 120 158 103

Exons Introns

Orthologs of CatSper3

OrganismHuman

Similarity


82.89(n)76.06(a)


99.66(n)99.25(a)

Cow(Bos taurus)

82.65(n)75.89(a)


73.72(n)67.83(a)

Mouse(Mus musculus)

74.43(n)66.67(a)


MAP LOCATION

Human : 1p35.3Mouse : 4D3Rat : 5q36

Start : 26,389,706 bp from pterEnd : 26,401,620 bp from pterSize : 11,915 basesEXONS : 10 ORF : 1419 nt Protein : 472 amino acids


I II IV V VI VIII IX

213 144 102 98 121 134 175 212 166 54

5’UTR 3’UTR


OrganismHuman

Similarity


82.76(n)77.12(a)


99.64(n)99.78(a)

Cow(Bos taurus)

83.88(n)78.51(a)


79.15(n)76.85(a)

Mouse(Mus musculus)

77.78(n)72.9(a)

LACK OF LITERATURE ON

ANY CATTLE (Bos spp)

Project executed in our lab

Title : Identification of SNPs in CatSper gene and their association with sperm motility in cattle

65

Diagram of Exon1-5 for Catsper1 gene on cattle

E-5E-3E-2 E-4E-1 E-15

10 kb

376bp 237bp282bp385bp299bp

66

Primers Designed (NC_007330) for SSCP analysis of CatSper1 Gene in Cattle

Sl No Primer Name Primer Sequence Primer Length (mer)

Product Length (bp)

1. Cat1-E1F 5'AGTGGAAGCGCACAGTCCTA3' 20 mer 299 bp

Cat1-E1R 5'AGGGATGGACCCTAATGGAG3' 20 mer

2. Cat1-E2F 5'GGCCCATGTGTTAAGCTTTC 3' 20 mer 376 bp

Cat1-E2R 5'ATCCTGGGAAAGGGATGTG 3' 19 mer

3. Cat1-E3F 5'CAGAAGGCCTACCTCCATGA3' 20 mer 237 bp

Cat1-E3R 5'AAGACCGCTGGACGAGAATA3' 20 mer

4.Cat1-E4F 5'GGGGAGTACCGTCATGGAAG3' 20 mer

385 bpCat1-E4R 5'GACTACACCAGCAGGGGAGA3' 20 mer

5.Cat1E5F 5'CCTTTCTGGCCCCCTTACA3' 19 mer

282 bpCat1E5R 5'ACCAACATCAACGGCCTTCTCTAC3' 24 mer

67

Optimized Conditions for PCR-SSCP Analysis

Gel Conc. : 12%

Time : 100bp/hr.

Temp. : 150C

Voltage : 350 Volts

Band patterns were resolved by Silver staining of

the gel (Bassam et al., 1991)

68

• Identification of SSCP band patterns

• Sequencing of SSCP patterns

• SNP identification by Nucleotide sequence analysis using MEGALIGN module

of DNA Star (Lasergene , USA) software.

• Estimation of Gene and Genotype frequency

Falconer and Mackey (2009)

• Haplotyping for SNPs data of each bull

• Anova using SAS programme (GLM, SAS 9.2) for ascertaining effect of

haplotype and its association with seminal parameters

• Statistical Model:

Yij = µ+Hi+eij

Where Hi is the effect of ith haplotype

µ = mean, eij =error effect

SNPs Identification and Association Study

69

Cattle Genomic DNA isolation

Conditions: Agarose gel 0.8%, 50 Volts for 1hr.

OD Ratio at 280/260: 1.6 to 1.9

Working DNA Concentration: 50-100ng/µl

70

PCR-Amplification for Catsper1 exon1

299bp

100bp ladder

S. No. Steps Temp. Time

1. Initial denaturation 95 °C 4min

2. 35 cyclesCyclic denaturation 94 °C 45 sec

Cyclic annealing 64°C 45 sec

Cyclic extension 72 °C 45 sec

3. Final extension 72 °C 10min

4. Storage 4°C 10min

71

SSCP pattern (monomorphic) in Catsper1 Exon1 (299bp)

Pattern frequency (zz)=100%

zz zz zz zz zz zz zz zz zz zz zz zz zz zz zz

72

PCR-Amplification for CatSper1 exon2

376bp

100bp ladder



2.35 cycles

Cyclic denaturation 94 °C 45 sec

Cyclic annealing 62.5°C 45 sec




73

SSCP patterns in Cat1 Exon2

AA (97)70.30 %, AB (16)18.10%, BB(25)11.60%

AA AA AB AA BB AB AA AA AA AB AA BB AB AA

74


237bp

100bp ladder



2.35 cycles

Cyclic denaturation 94 °C 45 sec





75

SSCP patterns in CatSper1 Exon3

DD DD EE FF EE GG GG HH

DD(58)42.03%, EE(34)24.64%, FF(25)18.12%, GG(9)6.52%, HH(12)8.69%

76


385 bp

100bp ladder



2. 35 cyclesCyclic

denaturation94 °C 45 sec





77

SSCP patterns Cat1 Exon 4

II X JJ JJ KK KK JJ LL

II(4)2.89%, JJ(101)73.19 %, KK(10)7.25%, LL(23)16.67%

78


282bp

100bp ladder



2. 35 cyclesCyclic

denaturation94 °C 45 sec





79

SSCP Patterns Cat1 Exon5

MM MM MN MN MN MN MN NN X NN NN NN NN NN

MM(21)15.22% MN(55)39.85% NN(62)44.93%

80

Total 10 novel SNPs identified in CatSper1 gene in cattle

Fragment 1

Fragment 2 Fragment 3 Fragment 4 Fragment 5

NO SNP T C, 199*(exon2)

C G 37(Intro1)

A G, 52 (Intron3)

G C 32 (exon5)Val TO Leu

G A, 272 (exon2)

Gly TO Ser

C T, 94 (exon3)

Thr TO Met

A G, 160 (exon4)

Thr TO Ala

T G, 165 (Intron3)

T C, 345 (Intron4)

G A,188 (Intron3)

* Silent mutation

81

Table: Fragment wise Gene and Genotype frequency for catsper1 gene in cattle Fragment No. Genotype Frequency % Allele Frequency %

Fragment 1 ZZ (138) 100 Z 100

Fragment 2 AA (97) 70.30 A 79.35

AB (16) 18.10 B 20.65

BB(25) 11.60

Total 3 (138) 100 2 100

Fragment 3 DD(58) 42.03 D 42.03

EE(34) 24.64 E 24.64

FF(25) 18.12 F 18.12

GG(9) 6.52 G 6.52

HH(12) 8.69 H 8.69

Total 5 (138) 100 5 100

Fragment 4 II(4) 2.89 I 2.89

JJ(101) 73.19 J 73.19

KK(10) 7.25 K 7.25

LL(23) 16.67 L 16.67

Total 4 (138) 100 4 100

Fragment 5 MM(21) 15.22 M 35.14

MN(55) 39.85 N 64.86

NN(62) 44.93

Total 3 (138) 100 2 100

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82

Haplotype analysis covering five exons of Catsper1 gene in crossbred bulls

Sl.No. Bull No.

Exon1 Exon2 Exon3 Exon4 Exon5 Haplotype

Pattern Pattern

Gen Pattern

Gen Pattern

Gen Pattern

Gen

1 67 ZZ AA G/G FF C/C JJ A/A MM C/C I-GC

2 70 ZZ AB G/A DD T/T JJ A/A MM C/C II-(G/A)T



5 969 ZZ BB A/A FF C/C JJ A/A MM C/C III-AC



83

Seminal Parameters Evaluation

Bull No. N Vol.(ml)

Conc (mill/ml)

MM(0-5)

IPM(%)

PTM(%)

(1) 70 V 21 3.95 ± 0.23 534.66 ± 52.10 3.47 ± 0.14 68.57 ± 2.41 43.33 ± 1.63

(2) 89 V 12 3.65 ± 0.28 533.91 ± 50.08 3.41 ± 0.19 72.91 ± 3.28 45.83 ± 2.20

(3) 1034 M1 18 4.17 ± 0.23 805.77 ± 49.63 3.94 ± 0.20 73.88 ± 2.27 48.88 ± 1.96

(4) 992 M1 12 4.05 ± 0.27 655.66 ± 47.62 3.41 ± 0.28 69.58 ± 3.45 45.83 ± 2.02

(5) 402 V 11 2.90 ± 0.33 671.54 ± 29.97 2.36 ± 0.15 61.36 ± 2.53 37.27 ± 1.56

(6) 969 V 14 4.63 ± 0.29 382.71 ± 39.87 1.21 ± 0.11 27.14 ± 2.26 12.50 ± 1.36

(7) 67 V 9 4.22 ± 0.27 566.66 ± 25.70 2.33 ± 0.16 53.33 ± 2.04 32.22 ± 2.22

84

ANOVA for Mass Motility

Source DF Sum of Squares

Mean Square F Value

Pr > F

Haplotype 2 52.5557429 26.2778715 35.07** <.0001

Error 94 70.4339478 0.7492973

Corrected Total

96 122.989690

85

ANOVA for Initial Progressive Motility


Mean Square

F Value

Pr > F

Haplotype 2 19904.24545 9952.12273 75.88** <.0001

Error 94 12328.74424 131.15685 Corrected

Total96 32232.98969

86

ANOVA for Post Thaw Motility


Mean Square

F Value

Pr > F

Haplotype 2 11329.65442 5664.82721 79.97** <.0001

Error 94 6659.00538 70.84048

Corrected Total

96 17988.65979

87

Association of bull fertility traits with possible haplotypes

HaplotypeSeminal parameters

MM IPM PTM

I-GC 3.22a ± 0.10

67.66a

±1.4543.22a ±1.06

II-(G/A)T 3.47a ± 0.18

68.57a

± 2.4943.33a

±1.83

III-AC 1.21b ± 0.23

27.14b ± 3.06

12.50b

± 2.24

88

CONCLUSIONSCONCLUSIONS

Five exons of catsper1 gene were PCR amplified in

cattle

A total of 15 SSCP patterns were found

10 novel SNPs were identified

3 haplotypes in bulls

Haplotype 1 and 2 significantly contributed to high

sperm motility

89

FUTURE PROSPECTS

The association of Identified haplotypes with sperm

quality traits still need to be validated with large

number of bulls and that could be ultimately used for

Marker Assisted Selection (MAS).

Genetic Effects of the identified DNA alteration at the

genomic level need to be confirmed at the

transcriptional or translational level.

90

Thank you

genomic insight of__sperm_motility

Health & Medicine

sperm flagellar motility

spag6 sperm

introduction sperm motility

mouse sperm

poor motility

motility defects

nonmotile sperm

ejaculated sperm flagellum