relationships between motility, fertility and cellular characteristics of spermatozoa, and the ability of

152 Pages·2017·1.07 MB·English

Clemson University
TigerPrints
All Dissertations Dissertations
5-2007
RELATIONSHIPS BETWEEN MOTILITY,
FERTILITY AND CELLULAR
CHARACTERISTICS OF SPERMATOZOA,
AND THE ABILITY OF PLATELET-
ACTIVATING FACTOR TO INFLUENCE
PORCINE PRODUCTIVITY
Jennifer Mook
Clemson University, [email protected]
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SPERMATOZOA, AND THE ABILITY OF PLATELET-ACTIVATING FACTOR TO INFLUENCE PORCINE
PRODUCTIVITY" (2007).All Dissertations. 84.
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RELATIONSHIPS BETWEEN MOTILITY, FERTILITY AND CELLULAR
CHARACTERISTICS OF SPERMATOZOA, AND THE ABILITY OF
PLATELET-ACTIVATING FACTOR TO INFLUENCE
PORCINE PRODUCTIVITY

A Dissertation
Presented to
the Graduate School of
Clemson University

In Partial Fulfillment
of the Requirementsfor the Degree
Doctor of Philosophy
Animal Physiology

by
Jennifer Leigh Mook
May 2007

Accepted By:
Dr. Ashby B. Bodine II, Committee Chair
Dr. John R. Diehl
Dr. William C. Bridges
Dr. James C. Morris
Dr. Lesly Temesvari

i
ABSTRACT

This research had 3 goals. First, differences between subjective and
objective motility analysis was determined using bull semen samples. Total
motility (MOT) of bull semen samples (N=117) was measured three ways:
subjective analysis manually on a microscope and objective analysis on 2
different computer-assisted semen analysis (CASA) systems: Hamilton-
Thorne IVOS and Minitube SpermVision (SV). CASA analysis also measures
progressive motility (PRO) and related kinematic parameters. Bull semen
motility analysis indicated a difference (p<0.05) between average MOT: 83.06
(IVOS), 75.84 (manual), 65.48 (SV), and PRO: 44.25 (IVOS) and 60.07(SV).
There were positive relationships (p<0.0001) between manual MOT and both
IVOS MOT (r=0.32) and SV MOT (r=0.59). The relationship between IVOS
and SV MOT and PRO was also notable (r=0.70 for both MOT and PRO).
Second, the ability of platelet-activating factor (PAF) to improve litter
survival parameters and fertilization/ embryonic death rates were assessed by
adding PAF (1x10-7M/mL) to extended high/low fertility boar semen prior to
artificial insemination of gilts synchronized with MATRIXTM. Gilts either
farrowed (N=67) or were slaughtered at day 30 of gestation (N=28). PAF
treatment of boar semen did not improve litter productivity parameters.
Notable variation in litter parameters was observed with low fertility semen
use. Gilt weight at time of insemination and boar fertility were the primary
factors influencing litter parameters and fertilization/embryonic death rates.

iii
Semen motility did not have an effect on litter parameters. Season did not
impact litter parameters when high fertility semen was used, but use of low
fertility semen in the spring resulted in lower (p<0.05) litter sizes.
Third, two cellular features that have been associated with fertility, PAF
receptor (PAFr) and ubiquitin (UBI), were quantitatively measured in
formaldehyde-fixed samples via fluorescence on a FACScan Analyzer
(Becton Dickinson) in bull (N=95) and boar (N=26) semen samples to
evaluate their relationship to motility and pig litter parameters. Boar semen
motility was measured on an IVOS. Analysis of cellular parameters showed
that increasing UBI was inversely related to CASA MOT and PRO of bull but
not pig samples. Increasing PAFr was associated with reductions in bull but
not pig PRO. There was no discernable relationship between UBI and boar
fertility, but there was a positive relationship (p<0.05) between UBI in
samples not washed with a modified Tyrode’s albumin/lactate/pyruvate
(TALP) solution prior to fixation and litter parameters: number born total
(r=0.55), number born alive (r=0.59), average litter weight (r=0.62), and
average wean weight (r=0.60). Washing boar samples with TALP increased
measurable levels of PAFr and UBI fluorescence (p<0.05). The data indicate
that UBI may be related to features associated with boar fertility, but more
work is needed to elucidate the effect of washing on overall fluorescence.
Treatment of boar semen with PAF did not impact gilt productivity as
expected, but future work needs to evaluate treatment concentrations and
interaction of PAF with extender components.
iv
DEDICATION

I dedicate this dissertation to four groups of people who have provided
tremendous support, guidance, encouragement and entertainment while I have
worked on this degree.
First, I would like to acknowledge the Atkins family: Michael, Linda,
Major and Chris. You have been my extended family in South Carolina and
have provided the support and encouragement that my immediate family
could not have provided on a daily basis. Without your love, this endeavor
would not have been possible. Saying that, the next group of people I
acknowledge is my family. Everyone in the Mook and Schaef families has
encouraged me and pushed me my entire life to reach for the stars. I hope I
have made you all proud. Third, I(cid:146)d like to thank Dr. Albert Abbott and the
Abbott Lab crew for supporting me and helping me through all the years I(cid:146)ve
been at Clemson. You have all been valuable friends. Lastly, I would like to
thank my (cid:147)boys(cid:148): John Cummings, Brad Robinson and Tafadzwa Kaisa, my
lunch time companions who put up with my disjointed communication skills,
my entertainment tidbits and my notable lack of skill in telling jokes. Thank
you all for being able to put a smile on my face when I needed one. To John,
thank you especially for making sure I had a job when I needed one and for
the general guidance and advice I have needed through the years. You are an
amazing person with one of the greatest minds I have ever encountered.

v
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ACKNOWLEDGEMENTS

Financial support for this work included the Wade Stackhouse Graduate
Fellowship as well as grants from the SC Pork Board, DANBREDUSA, and the

California State University Agriculture Research Initiative (ARI).
Many people contributed their assistance to the project including: Sandra Gray
(cid:150) TALP recipes and supplies, Nancy Korn (cid:150) lab techniques, Dr. John Gibbons (cid:150)
lab equipment and chemicals, Dr. Steve Ellis (cid:150) lab assistance and chemicals, Dr.
Plummer and his students (cid:150) BSE evaluations and semen collections at the
California Polytechnic University, San Luis Obispo, CA, Kathleen Baska and
Peter Sutovsky (cid:150) bull semen processing and flow cytometry sample analysis, Dr.
Charles Francisco - MATRIXTM donation from Intervet, Hamilton-Thorne/Nancy
Smith/Kate M. Craig - donation of IVOS and IVOS operation at BSE evaluations,
Minitube/Brian Day (cid:150) donation of SV and SV operation at BSE evaluations,
Greenville Hospital System, Department of Reproductive Endocrinology and
Infertility/Dr. William Boone/Dr. Lee Higdon/Glenn Proctor/Jennifer Graves (cid:150)
Use of IVOS and related materials, and the Clemson University Starkey Swine
Center Staff(cid:150) maintaining farm facilities/livestock and general assistance with
research. Lastly, I(cid:146)d like to thank Dr. Diehl, Dr. Bodine, Dr. Bridges, Dr. Morris
and Dr. Temesvari for serving on my committee.

vii
viii
TABLE OF CONTENTS

Page
TITLE PAGE(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) i
ABSTRACT(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. iii
DEDICATION(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. v
ACKNOWLEDGEMENTS(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. vii
LIST OF TABLES(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) xi
LIST OF FIGURES(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. xiii
CHAPTER
1. LITERATURE REVIEW(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 1
a. Search for fertility factors(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) 1
b. Bull and boar fertility(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)... 4
c. Platelet-activating factor (PAF)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)... 8
i. Biochemistry of PAF(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).... 8
ii. PAF and cell signaling(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 11
iii. PAF in reproduction(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 13
d. Ubiquitin(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. 16
e. Computer assisted semen analysis(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)... 20
f. Flow cytometry(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).... 28
2. RELATIONSHIP OF PLATELET-ACTIVATING RECEPTOR
(PAFr) AND UBIQUITIN, AS FERTILITY INDICATORS IN
THE BULL, TO MOTILITY PARAMETERS DETERMINED
VISUALLY AND ON TWO DIFFERENT COMPUTER-
ASSISTED SEMEN ANALYZERS(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)..(cid:133).. 31

a. Introduction(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. 31
b. Materials and methods(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. 33
c. Results(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. 39
d. Discussion(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 49

ix
Table of Contents (Continued)

Page

3. RESULTS OF ADDING PLATELET-ACTIVATING FACTOR
TO LOW AND HIGH FERTILITY BOAR SEMEN PRIOR TO
ARTIFICIAL INSEMINATION ON LITTER PRODUCTIVITY
OF GILTS SYNCHRONIZED WITH MATRIX(cid:153)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) 59

a. Introduction(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. 59
b. Materials and methods(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 62
c. Results(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133).. 70
d. Discussion(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) 87

4. CONCLUSION(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) 95
APPENDICES(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) 101
a. TALP Solutions(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133) 103
b. Basic data for each gilt used(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 105
c. Farrowing and fertilization/embryonic death results
for all gilts(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 109
d. All results for Table 15(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 113
e. All data for Table 17(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133). 115
f. Boar information(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)... 117

LITERATURE CITED(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)(cid:133)... 121

x