Advances in Bull Fertility and Semen Evaluation From Sperm Phenotype to Genome and Back
Peter Sutovsky, PhD, Dr.h.c. Division of Animal Sciences, and Departments of Obstetrics, Gynecology and Women’s Health, University of Missouri, Columbia, MO 62511 [email protected] Thank You
Brazil 2015 TRUMAN Sutovsky Lab Intro
• Established 2001 • Male fertility in livestock and humans • Assisted Reproductive Technologies/Therapy (ART) • Improvement of Artificial Insemination (AI) Ubiquitination Targets Proteins for Degradation by The 26S Proteasome UPS in Gametogenesis, Fertilization & Development – USDA Projects 1998-2019
EPIDIDYMAL SPERM MAJOR ZYGOTIC MITOCHONDRIAL PRONUCLEAR MATURATION – GENOME INHERITANCE: Sperm DEVELOPMENT & HISTONE Defective sperm ACTIVATION mitophagy after MODIFICATION ubiquitination fertilization:
FERTILIZATION: Sperm-zona penetration & anti- polyspermy defense
SPERMATID DIFFERENTIATION, OVARY & OOCYTE - Oocyte SPERM CAPACITATION ACROSOMAL maturation, cortical granule Sperm remodeling in BIOGENESIS biogenesis, meiotic spindle preparation for function fertilization Guinea pig Ostrich
H. sapiens..?
Zebra finch Opossum Drosophila Large Animal Models
For every physiological problem, there will be some animal of choice, or a few such animals, on which it can be most conveniently studied.
(The Krogh Principle) Goals
• Better understand reproductive process in mammals
• Improve reproductive health, well being and increase the efficiency of assisted reproductive therapy in humans
• Increase the efficiency od reproductive performance in food animals (AI, IVF, ET, sexing, cryopreservation, reprogenomics) Market Need/Opportunity
Annual semen sales: $1.5 billion (dairy), $250 million (beef), $600 million (swine)
Trend & Hotspots: • Genomics • Sexed semen • Higher AI fertility • Maximized AI dose production from genetically valuable sires Problems & Solutions
• Suboptimal conception rates, early pregnancy loss • Lack of objective semen analysis methods • Mitigation of sperm damage from cryopreservation, sexing • Fertility biomarker discovery & validation • Improvement of AI dose • Maximize extension of semen from sires with valuable genomes outline
1. Brief overview of defective sperm phenotypes and their causes 2. Conventional and objective methods for semen analysis 3. Biomarkers of sperm quality/fertility 4. Biomarkers of sperm capacitation – Zinc signature 5. Genomics of sperm phenotype Teratospermia
• Generally poor morphology, motility, viability • Various causes – age, toxicants, heat stress, nutrition Causes of Reduced Sperm Quality
• Age • Health (infections) • Nutrition (malnutrition, trace element imbalance, toxicants) • Environment (heat stress, pollution) • Genetics • Damage from collection and processing (cryopreservation/cryocapacitation, sexing) Heat Stress
• May be partially, completely reversible • Cause variety of sperm defects • Can be studied by scrotal insulation Infectious Bacterial Disease
• Leucocytospermia • Increase in amorphous cellular debris • Semen quality may recover after antibiotic treatment • Case in point: ubiquitin and PAFR (WBC marker) levels recovered in yearling bulls treated with antibiotics
Sutovsky et al., J. Androl. 2006 Feed Toxicants
• Gossypol (cottonseed feed, Chenoweth 2000) • Decapitation, head defects, midpiece lesions, cytoplasmic droplet (CD) retention
Corkscrew
Nuclear crest
Proximal CD Distal reflex
Distal CD Genetic Causes
• Single nucleotide polymorphisms (SNP)/point mutations • Differences in copy number of multi-copy genes • Epigenetic aberrations • Early example: Dag defect • Often recessive, manifested only in homozygous configuration Heritable Sperm Defects
• Globozoospermia (round headed sperm syndrome) • Pyriform head • Knobbed acrosome • Fibrous sheath dysplasia (stump tail) • Multi-nuclear, multi- flagellar defect • Asthenozoospermia (flagellar defects) 2. Semen Analysis
First (Human) Sperm Observations Van Leeuvenhook & Hamm 1677-79 Conventional Semen Analysis 1909 A careful Microscopic examination is a valuable aid in determining the nature of chronical diseases of generative organs Spermatorrhea – seminal weakness*
*may be a result of marital excess University of Missouri 1917
Frederick B. Mumford 1868-1946 Conventional Semen Analysis 2019
• Front line semen assessment (volume, density, color, swirl) • Sperm Count • Motility • Appearance/Morphology • Contaminants (Leukocytes, spermatids, epithelial cells, residual bodies, cellular debris). • Not always correlated with field AI fertility Good or Bad?
Conventional Detection of “Good “Bad Sperm” Marker Semen Analysis Fertility” Protein PAFr Ubiquitin SOLUTION: Biomarker-based Sperm Quality/ Fertility Assays
• Biomarker-based, objective, automated • Correlate with conventional semen parameters and field AI fertility • Inexpensive, simple, and predictive of sires’ future fertility 3. Biomarker Identification by Comparing Normal vs. Defective Sperm Fraction Proteomes
1-13
Infertile Defective Normal
14-21
Fertile
Surface-Enhanced Laser Desorption/Ionization (SELDI) P62 UBA Negative Biomarkers of Male Fertility
• Vital stains (mitopotential, acrosome, live/dead)
• Ubiquitin (UBB)
• 15-lipoxygenase (ALOX15)
• Lectin ligands (LCA, PNA)
• PAWP (WBP2NL)
• Aggresome (AGG) Sperm Flow Cytometry
Flow Cell
Photo Laser Detectors Side Scatter
Forward Scatter Flow Cytometry – Histogram & Scatter Diagram
Normal Histogram Abnormal Histogram Image-based FC: Cytometer & Microscope in One Box
Normal Sperm (R3) Defective Sperm (R2)
Amnis Corp., Seattle WA [Buckman et al., 2009, Systems Biol. Reprod. Med., 55(5-6):244-251] Ubiquitin (UBB)
• Protein recycling peptide tags defective proteins, binds covalently to the surface of defective spermatozoa • • Ubiquitinated proteins of testicular origin may be carried over by defective spermatozoa Apocrine Secretion of Ubiquitin in the Epididymis
From Hermo and Jacks, 2002 UBC-GFP Rat (Baska et al., 2008) Defective Sperm Ubiquitination
Sutovsky et al., 2001, J. Cell Sci. 114:1665-75. Possible Roles of Defective Sperm Coating
• Clustering of defective spermatozoa
• Prevention of autoimmune infertility
• Degradation of defective spermatozoa Ubiquitin Correlates With PARAMETER r Breeding Soundness Parameters Scrotal circumference -0.79 in Yearling Bulls Color (1-4) -0.59 Semen Volume -0.42 BSE identifies young bulls satisfactory for Swirl (0-5) -0.64 breeding based on a physical examination, Strength of Motility (SMR; 0-5) -0.71 reproductive organ examination including % Progressive Motility -0.72 of scrotal circumference and semen analysis % Normal Sperm -0.73 % Head Abnormalities +0.65 USDA, Forth Keogh, MT (n=207 Yearlings) % Total Abnormalities +0.74
TABLE: Highest correlations* between sperm UBI and % Live Sperm -0.71 BSE parameters obtained in 5 flow cytometry UBI trials Hemocytometer dilution -0.37 testing a total of 207 bulls Concentration -0.56 Pearson’s r-coefficient (0.01-0.99) Contaminants +0.29 Ubiquitin Correlates with AI Fertility Indexes
%M2 (good MedM3 (Bad Sperm Med-All MedM1 MedM2 Sperm) %M1 ) %M3
Services, ATA 02-07 0.070911 0.241528 0.085717 -0.15057 -0.23675 0.402741 -0.13591
Services, ERCR 02-07 0.297884 -0.00446 0.279037 0.1108 -0.28891 0.171768 0.24182
Services, RBE II 01-07 0.267008 -0.16777 0.221209 -0.04858 -0.26394 0.082796 0.314891
DEV, ATA 02-07 -0.13288 0.244449 -0.10763 -0.50733 -0.07998 0.322993 -0.26919
DEV, ERCR 02-07 0.180263 0.40738 0.142119 -0.26655 -0.3231 0.351187 0.076113
DEV, RBE II 01-07 -0.09158 0.318927 -0.1193 -0.64229 -0.09412 0.222514 -0.09654
RBE II Index 01-07 0.07999 0.306967 0.03692 -0.58223 -0.32076 0.427711 0.011327
Services, CFI 02-07 0.267276 0.08448 0.245291 0.03589 -0.31763 0.26772 0.167393
DEV, CFI 02-07 0.008237 0.504845 -0.02311 -0.44093 -0.25192 0.425151 -0.12445
CFI Index, 02-07 0.11139 0.526457 0.079442 -0.47993 -0.34474 0.481186 -0.04369 • N=15 sires/85 collections Aggresomes (AGG)
• Proteinaceous inclusion bodies that form when the ubiquitin-proteasome machinery is overloaded (cellular stresses, ROS overproduction)
• Degraded by autophagosome, bind SQSTM1
• Sperm mitochondrial sheath has aggresome-like properties
• Detected by ProteoStat Aggresome Detection Kit (Enzo), a 488 nm excitable molecular rotor dye that binds to denatured proteins Does Size Matter?
HYPOTHESIS: Length of sperm tail mitochondrial sheath (MS) correlates with conventional semen and sperm parameters, and with fertility in bulls used for artificial insemination (AI) service.
Fit Plot for Pregnancy and Fit Plot for Fertility Index and
Mitochondrial Sheath Length Mitochondrial Sheath Length
Fertility Index Fertility Pregnancy Rate (%) Rate Pregnancy
Expected Progeny Difference (EPD) Correlations
Mitochondrial Sheath (MS) Length Reproduction Correlations Avg MS Length:Yearling Weight -0.61
Nelore 9.74±0.31 µm Avg MS Length:Bull Fertility Index 0.38 Avg MS Length:Weaning Weight -0.59
Angus 9.35±0.28 µm Avg MS Length:% Live Sperm 0.41 Avg MS Length:Residual Average -0.50 Daily Gain P<0.0001 Avg MS Length:Motility 0.04 Avg MS Length:Calving Ease -0.61 Std Dev MS Length:% Dead and 0.44 Modified Sperm Acrosome Avg MS Length:Maternal Milk EPD -0.63 Grace Wiley, Eriklis Nogueira, Camile Sanches, Karl Kerns, Peter Sutovsky Avg MS Length:Fat Thickness EPD -0.58 Future is in Multiplexing
Kennedy et al., 2014, Mol Reprod. Dev. 81:436-449 Ubiquitinated Spermatozoa Lack PAWP Protein Postacrosomal Sheath WW-Domain- Binding Protein (WBP2NL/PAWP)
• Resides in the post-acrosomal sheath of the sperm head perinuclear theca (PT)
• Promotes oocyte activation and pronuclear development during fertilization
• Present in normal sperm but abnormal sperm may have elevated levels of PAWP PAWP-Candidate SOAF*
Sperm-borne Oocyte Activating Factor
Before sperm- oolemma fusion
After sperm- oolemma fusion
Wu et al., 2007, J. Biol. Chem. 282:12164-75. Aarabi et al., 2014, FASEB J PMID: 28(10):4434-40 Gating of Low, Medium and High PAWP in Bull Spermatozoa
Low/no PAWP Medium/Optimal PAWP High/Excess PAWP Sperm Phenotype Analysis by Image-based FC
Spermatozoa with low/no PAWP are most likely to have sperm head defects
Kennedy et al., 2014, Mol. Reprod. Dev. 81:436-449 Ubiquitin Binding Probes Recognize Spermatozoa with Hidden Abnormalities, DNA Damage
Sutovsky et al., 2002, Mol. Reprod. Dev. 61: 406-413 Semen Content of Ubiquitinated Spermatozoa Correlates with Sperm Acrosomal Damage
• Essential for fertilization process
• Induced by sperm binding to sperm receptor protein on the ZP
• Irreversible Peanut Agglutinin (PNA)
• High affinity/strong specificity for disaccharides with terminal galactose, especially the D-Gal-α(1,3)-D GalNAc disaccharide
• Binds to the outer acrosomal membrane (OAM), exposed during capacitation and acrosome reaction Dual UBB-PNA Assay
Odhiambo et al., 2011 Lentil Lectin (LCA)
• Shows a strong specificity to d-glucosyl and d- manosyl residues • Binds to the acrosome in normal sperm, whole surface in defective sperm Bull Fertility Biomarker Trial - Results
UBI vs. LCA • Ubiquitin is positively 60 correlated with high % 50 LCA labeled
40 spermatozoa
30
LCA(Total) %M7 • r=0.65 20
10
0 0 5 10 15 20 25 30 35 40 45 UBI % M3 UBI (Total) Multiplex Assay Correlations
Kennedy et al., 2014, Mol Reprod. Dev. 81:436-449 Ubiquitin-PNA Based Semen Nanopurification
• Goal: Remove defective spermatozoa from semen used or artificial insemination (AI)
• Benefits: Increased fertility/pregnancy rate, lower dose of spermatozoa per insemination, efficient use of genetically valuable sires, possibly eliminating the need for semen pooling
Sperm Depletion Protocol
15 min incubation
15 min. magnetic particle pull down
Cryopreservation
Artificial Insemination Semen nanopurification
Defective sperm removed
Normal sperm retained 466 Healthy Calves Born to 798 Inseminated Cows & Heifers
100% Pregnancy Rates of Semen Treatments 90%
80%
70% 64% 65% 60% 54% 51% 50%
40%
30%
20%
10% n = 228 n = 240 n = 160 n = 170 0% 10M Control 20M Control 10M UBI 10M PNA
Odhiambo et al., 2014, Biol. Reprod., Semen Nanopurification in Other Species
Domestic Pig AI Human Infertility Therapy
https://www.youtube.com/watch?v=ZsFsyFQTljs Feugang et al., 2015, JFIV http://www.mcrmfertility.com/treatment-options/in-vitro-fertilization-ivf- Reprod. Med. Genet. 3:145 /selecting-the-perfect-sperm/nanobead-sperm-selection-process.aspx CONCLUSIONS - BIOMARKERS
• Normal and defective spermatozoa have different proteomes and ubiquitomes
• Sperm levels of “negative” fertility markers correlate negatively with conventional parameters of semen quality and fertility in bulls and boars
• Negative biomarkers on the sperm surface can be targeted by semen nanopurification to improve AI fertility and reduce sperm number per AI dose 4. Markers of Premature Sperm Capacitation
• Suboptimal handling and extenders after collection • Time between collection and processing • Cryocapacitation • Semen sexing-induced capacitation
Redgrove 2012 Gamete Transport & Fertilization
1. Deposition of spermatozoa & sperm-protective seminal plasma
2. Acquisition of motility & rapid swimming through cervix removes seminal plasma
3. Sperm movement through uterine cavity assisted by muscle contractions
4. Slowing of the sperm in uterotubal junction or isthmus
5. Binding to oviductal epithelium (sperm reservoir)
6. Capacitation, hyperactivation and sperm release induced by ovulation
7. Oocyte picked by cilia on fimbria, moved down ampula by cilia and contractions
8. Fertilization occurs in ampullary-isthmic junction (Suarez, 2002, 2008) Oviductal Sperm Reservoir
• Binding of spermatozoa to oviductal epithelium in utero-tubal junction or isthmus • Described first by Yanagimachi and Chang (1963) • Present in cows, pigs, hamsters, sheep, hares, mares (human???) • Spermatozoa bind to fucosylated ligands resembling Lewis trisaccharide on the surface of oviductal epithelium (Annexins)
Proposed Functions: • Maintenance of spermatozoa between the onset of oestrus and ovulation • Synchronization of sperm & oocyte transport in the oviduct
Release: • Sperm capacitation & hyperactiovation • A change it the sperm surface, rather than a change in oviductal epithelium • Only hyperactivated spermatozoa can detach Set of changes in the sperm plasma Sperm membrane that enables a cell to acquire fertilizing potential/undergo Capacitation – acrosomal exocytosis
Point of No Asynchronous and continuous: Only a percentage of spermatozoa are capacitated Return and those are continuously replaced
Probably triggered by oviductal secretion near the time of ovulation
Requires removal of seminal plasma -SP has decapacitating activity
M.C. Chang (1951) & C. R. Austin (1951) observed that rat Capacitated state lasts 50-240 minutes. spermatozoa cannot penetrate the Post-capacitated cells die unless they oocyte immediately after coitus and need ~2 h in the female tract undergo acrosome reaction. to acquire such an ability. Sperm Capacitation Outcomes
• Formation of membrane rafts • Plasma membrane & acrosome remodeling • Protein phosphorylation increase • Calcium influx • Hyperactivation • Detachment from sperm reservoir
• Gadella & Visconti, 2006; In: The Sperm Cell, Cambridge University Press • Visconti et al., 1995, Development 121, 1129-1137 • Lishko et al., Sperm Capacitation Status
• Spermatozoa do not capacitate naturally until they bind to oviductal sperm reservoir epithelium • Premature capacitation kills spermatozoa • Associated with Ca-influx in sperm • Measured by fluorescent Ca-dyes Fluo-3/Fluo-4 (flow cytometry) or by chlorotetracycline (epifluorescence microscopy)
Cap. Non-cap. Zinc Spark
• Rapid efflux of ooplasmic zinc ions after sperm-oolemma fusion • Zinc similar to Ca is a signaling molecule, oocyte content of Zn increased during maturation (cortical zinc vesicles) • Decrease in Zn ions may be cell cycle related, necessary for progressions of second meiosis • Sen only after activation of MII, not GV oocytes • Role in anti-polyspermy defense, sperm alteration, repulsion, loss of chemoattractant?
Woodruff & O’Halloran Labs, Northwestern University Chicago, IL Zinc Chemorepels Capacitated Spermatozoa (mouse, rabbit, and human)
Progesterone Zinc
Guidobaldi et al., 2017 Human Repro If capacitation is defined solely as the capacity to fertilize, it would seem reasonable that the oocyte can de- capacitate spermatozoa via Zn2+ signaling. Hypothesis:
Zinc Spark
+ 'Zinc Shield' 300% Zona Pellucida Zn
Que et al., 2017 Zn2+ – The Capacitation Signature
• Quick one step live sperm staining, detects & Before quantifies premature capacitation capacitation • Also detects & quantifies death sperm
• Indicates sperm ability or readiness to undergo timely capacitation
After capacitation DEAD NON-CAPACITATED
CAPACITATED
Fluorescence Intensity Kerns E et al 2018 Nature Communications Kerns et al., unpublished Super Resolution Microscopy
Kerns et al., in preparation SBiRM Vol. 65, 2019 - Cover ART by Karl Kerns PhD Mammalian Sperm Zinc Signature
Kerns et al., 2018, Nat Comm Methodology: Image-based Flow Cytometry with FluoZin3 Probe Zinc Signature Altered with In Vitro Capacitation (IVC)
Kerns et al., 2018, Nat Comm Zinc Signature is Associated with Capacitation-induced Acrosomal Changes (PNA) and Plasma Membrane Permeability (PI)
n=4, P ≤ 0.001, 10,000 cells/treatment analyzed
Kerns et al., 2018 Nat Comm Does Semen Sexing Affect Zinc Signature?
• Same bull, same ejaculate • ½ neat, ½ XY-sorted
Control Sexed
(Zn2+ fluorescence in sperm head) Co-Management of Sperm Mitochondrial Health and Zinc Signature
(Fertilization)
Traditional Semen Handling
Managing the Sperm Zinc Signature
Capacitation Time Clock and Sperm Lifespan Seminal Plasma:
• Produced by male accessory sex glands, rich in Zn2+ • Semen extension dilutes Zn2+ ions in seminal plasma
• FUNCTIONS:
• Alkaline pH for neutralization of vaginal environment
• Induction of sperm progressive motility ( activates sperm soluble adenylate cyclase/SACE, elevates cAMP)
• Formation of vaginal /copulatory plug (rodents)
• Protease inhibitors, prostaglandins, growth factors, immuno-suppressors are present in seminal plasma
• Protective coating of sperm surface (spermadhesins/binder of sperm proteins)
• Induction of CD removal (boar)
• Induction of ovulation (alpaca, beta-nerve growth factor)
• Sperm surface-binding proteins required for sperm binding to oviductal sperm reservoir (e.g. BSPs in bull, spermadhesins in boar)
• Seminal plasma may influence gene expression in uterine epithelia directly and indirectly regulate embryo development through uterine secretion of embryotrophic growth factors (Bromfield et al., 2014, PNAS 111:2200-5.) MU Vanguard II – Day 21 Spermatozoa
0% motility 2%
Control MU Vanguard
• Uses Zn-management and synthetic forms of mito-protectants produced naturally by mitochondria of somatic cells • Sperm storage in dark, at room temperature MU Vanguard Extender - IVF Trials*
*48 h room temperature storage; pbts-muv=0.02 CONCLUSIONS – Zinc Signature
• Zinc ion efflux is a hallmark of sperm capacitation
• Premature capacitation can be mitigated by Zn- reloading
• Diets should be balanced for micronutrients such as Zn2+, specifically for AI bulls/boars
• Mito-protectant, Zn-enriched semen extenders could extend lifespan of extended semen, protect spermatozoa from damage during sexing and cryopreservation Kerns et al., Intl J Mol Sci, 2018
• Sperm Zn imaging is a tool for Androlab quality control (AI, cryo, sexing) 5. Linking Sperm Phenotype to Genome - Dual Purpose with Dual Benefit
• Joint USDA-NIH Initiatives
• One Health One Medicine
• Research in Biomedicine and Agriculture Using Agriculturally Important Domestic Animal Species
• Grant 5 R01 HD084353-02 Linking Fertility-Associated Gene Polymorphisms to Aberrant Sperm Phenotypes (Peter Sutovsky, Jeremy Taylor & Robert Schnabel Co-PIs) Haplotypes Associated With Bull Fertility • 255 AI sires with varied semen quality Cooperative Dairy DNA Repository aligned to NCBI sequences over 2,500 bull genomes
• Variants were called and the variant data were submitted to the 1000 Bull Genomes Project for inclusion in the global variant dataset which will be derived from approximately 4,000 genomes
• Master list of candidate single nucleotide polymorphisms (SNPs) affecting bull fertility will be used for construction of genotyping microarrays.
From Taylor et al., 2018, • Focus on rare homozygous recessive Animal 9:2061. mutations Identification of a Rare, Fertility Affecting Mutation in Bovine Eml5 Gene
• Echinoderm microtubule- associated protein-like 5 isoform X5 [Bos taurus] • Rare mutation in WD40 domain • Repetitive, circular solenoid protein domain for multi-protein complex assembly
Michal Zigo1, Eriklis Nogueira2.3, Karl Kerns1, Miriam Sutovsky1, JaeWoo Kim1, Filip Tirpak4, Thomas E. Spencer1,5, Jeremy F. Taylor1, Robert D. Schnabel1,6, Peter Sutovsky1,5 Distribution of EML5 in Bull Testis
• Between spermatocyte chromosomes • Inside spermatid nucleus • Around acrosomic granule Immunolocalization & Quantification of EML5 Protein in Spermatozoa of Wild Type and Eml5wd40+/- Mutant Bulls
Wild type +/-Mutant
Immunofluorescence Flow Cytometry Protruding Knobbed Acrosome Phenotype in Homozygous Eml5wd40+/+ Mutant Bull Distribution of EML5-related Protein EML4 (red) in Spermatozoa and Testicular Tissues
• Cytoplasmic lobe of EML4 Acrosome DNA round spermatids • Caudal manchette of elongating spermatids • Connecting piece & midpiece of normal spermatozoa • Aberrant localization & high accumulation in mutant & abnormal spermatozoa Wild type (WT) Eml5 Stump tail mutant WT Conclusions – Genomics of Sperm Phenotype
• Carrier bulls with sperm-affecting SNPs are rare within the AI pool and may have compromised fertility, which may be tolerated by the AI industry due to their high genetic value for production traits.
• Identification of fertility affecting mutations is important for cattle genomic selection and reproductive management, and significant for the diagnostics and therapy of human infertility.
• Data from this project will be used to build a bull fertility microarray. OVERALL IMPACT
• Improved methodology for semen processing, including extension of fresh semen shelf life and improved viability after cryopreservation. • Improved commercial embryo transfer by promoting optimal pronuclear development, timely MZGA and high rate of development to blastocyst. • Mitigate early pregnancy loss which is impacting the economics of cattle and swine breeding • Advance basic knowledge of reproduction and development
Kerns & Sutovsky, Syst Biol Reprod Med Cover Figure, 2019 IMPACT Ctd’
• Taking full advantage of new trends in precision agriculture (genomic selection, semen sexing) • Lowering the cost of AI by reducing sperm number/dose • Reducing the environmental impact of AI studs (fewer bulls needed) • Healthy Bottom Line = Food $ecurity MU Collaborators Lab-Present External Research Collaborators Miriam Sutovsky Randy Prather MU ANSCI Karl Kerns Kevin Wells MU ANSCI Bill Ballard, Sydney, AUSTRALIA Jerry Taylor & Bob Schnabel MU ANSCI & GENOMICS Justin St John, Hudson Institute, AUSTRALIA Michal Zigo Tim Safranski MU ANSCI Mark Baker & John Aitken, Newcastle, AU Dalen Zuidema Gavin Conant MU ANSCI Stuart Meyers, UC Davis Grace Wiley Jamal Ibdah – MU School of Medicine Richard Oko-Queen’s U. Kathy Sharpe-Timms MU-OBGYN Tom Geary USDA-ARS Kathy Craighead Gary Clark MU-OBGYN David Miler, U. of Illinois Laura Schultz MU OBGYN Maik Huttemann, Wayne State, Detroit, MI Yuksel Agca MU VETMED Markéta Sedmíková, Lucie Tumová, CZU Heide Schatten MU VETMED Lab-Past (CULS ) Prague Dietrich Volkmann MU VETMED Vӗra Jonáková , Jana Pӗknicová , Eriklis Nogueira Susan Nagel, MU OBGYN Pavla Postlerová, Inst. Biotech., Prague Filip Tirpak Jan Nevoral, Miriam Štiavnická, Charles Seda Ocakli University in Pilsen CZ Lucie Tumova Katerina Dvoraková-Hortová, Charles U., Miriama Stiavnicka Prague Clio Maicas Eva Nagyová, Andrej Šušor, IAPG, CAS, Jiude Mao Libĕchov, CZ Keith Latham, Temple U. Jenny Jankovitz Kingston, ON Wonhee Song Erdogan Memili, U. Linda Yan Mississippi Jan Nevoral Namdori Mtango, Peter Petruska Temple/Minitube Veronika Benesova Susan Suarez, Cornell U. Young-Joo Yi Satish Gupta, Natl. Inst. Immunol., New Delhi TJ Miles Antonio Miranda-Vizuete, Peter Vargovic UPdO, Sevilla Shawn Zimmerman Klaus van Leyen, Harvard Chelsey Kennedy Winston Thompson, Morehouse School Gauri Manandhar of Medicine Fred Odhiambo Fred van Leeuwen, University of Maastricht T.J. Myles Katie Fischer Current Funding Kathleen Baska USDA-NIFA NIH-USDA Dual Purpose - NIH-NICHD Kyle Lovercamp F21C UM Kelly Moore Dawn Feng Industry Collaborators Jen Antelman Mel DeJarenette-Select Sires Heinz Leigh Kari Beth Krieger – Genex Nicole Leitman Abdullah Kaya -Alta Clayton Buckman Tom Geary, USDA Eric Schmitt, IMV Technologies Peter Ahlering, MD, MCRM, St. Louis
The Great Facebook Caption Challenge
Infertility Tango Hooking up? Tug of (sperm) war Twisted Sister before the band got together A tale of two sperm: “It was the best of sperm it was the worst of sperm” Yours?