Animal Science 434

1. Animal Science 434 Epididymis, Ejaculation and Semen

2. Head Sperm Structure Acrosome Neck Middle Piece Annulus Principal Piece End Piece

3. Sperm From Different Species

4. Sperm Head (Equatorial Segment) Nucleus Nucleus

6. Acrosome Plasma Membrane Nucleus

7. Acrosin • Hyaluronidase Acrosome Nucleus Plasma Membrane

8. Posterior head Capitulum Laminated columns(flexibility) Mitochondrial helix (energy) Middle piece Axoneme (9 + 9 + 2) Course outer fibers

9. Annulus Midpiece Principal Piece Fibrous Sheath or helix

11. Axoneme Principal Piece Middle Piece

12. Axoneme

13. Microtubule Sliding in Sperm Tail

14. Sperm Metabolism Hexokinase Glucose Glucose-6-phosphate Phosphohexoisomerase Anerobic Metabolism (Glycolysis) Fructose-6-phosphate Fructose Hexokinase Lactate + 2 ATP + 2 H2O Fatty Acids Aerobic Metabolism Tricarboxcylic Acid Cycle 36 ATP/Glucose 36 ATP/Fructose

15. Sperm Metabolism phosphatase Hexokinase Glucose Glucose-6-phosphate Phosphohexoisomerase Anerobic Metabolism (Glycolysis) Fructose-6-phosphate Fructose Hexokinase Lactate + 2 ATP + 2 H2O Fatty Acids Aerobic Metabolism Tricarboxcylic Acid Cycle 36 ATP/Glucose 36 ATP/Fructose

16. ATP Utilization in Sperm • motility (60%) • substrate cycling (40%) • wasted • maintenance of ionic gradients • very small amount • transcription and translation • none after condensation of nucleus and loss of residual body

17. Sperm Metabolism • Temperature dependent • ATP production increases as temperature increases • ATP dependent processes are temperature dependent • motility increases with increasing temperature

18. Effect of Temperature on Motility

19. Transport of Sperm Vas Efferentia • fluid flow • cilia Caput • contractions Vas Deferens Rete Testis • contractions • fluid flow • RT secretion Corpus • contractions Seminiferous Tubule • bulk fluid flow • contractions ofof myoid cells Cauda • contractions

22. Epididymal Function • Maturation • change in fertility • develop motility • nuclear condensation • cytoplasmic droplet • Concentration • remove water

23. Epididymal Function • Secretion • T DHT • energy substrates (GPC, free fatty acids, carnitine) • glycoproteins, lipids, enzymes • Transport • smooth muscle contraction • Storage • cauda

24. Hormonal Control of Epididymis • Caput • tubal testosterone • Corpus • tubal dihydrotestosterone (DHT) • vascular testosterone • Cauda • tubal DHT • vascular testosterone • Absolute need for Androgen

25. Erection Sensory Psychic Sympathetic relaxation of penile retractor muscle Parasympathetic contraction of ischiocavernosus muscle dilation of vessels Inhibit vasoconstriction engorgement of corpus cavernosum decreased venous outflow

26. Emission and Ejaculation Sensory Psychic Parasympathetic Sympathetic smooth muscle contraction of prostate, seminal vesicles, vas deferens, cauda epididymis, closure of the sphincter to the bladder striated muscle contraction of bulbospongiosus, pelvic urethral emission ejaculation

27. Ejaculate Characteristics Species Time lapse for Composition of Bull 1 second single fraction Ram 1 second single fraction Boar 5 - 25 minutes fractionated sperm-free sperm-rich coagulum Stallion 30 - 60 seconds fractionated sperm-free sperm-rich mucus Human 10 - 30 seconds single fraction but coagulated

28. Components of Semen • water • sperm • substrates • fructose • sorbitol • inositol • glycerol phosphatidyl choline (GPC) • citric acid • inorganic salts • sodium, potassium, calcium, magnesium, chloride • proteins

29. Initial Semen Evaluation • color • volume • concentration • motility • viability • morphology

30. Semen Color and Volume • Color • White to cream color - good • Yellow – urine present • Clear or thin in appearance – few sperm • Volume • Weight, 1 ml = 1 gm • Concentration • Hemocytometer – gold standard • Spectrophotometer, photometer, fluorometer

31. Motility Evaluation • Also a viability evaluation • Visual estimation – Nearest 5% estimate • Total motility – all the moving sperm • Progressive motility – those sperm moving in a straight line • CASA – Computer aided sperm analysis • Tracts individual sperm and computes a variety of statistics on the sample • Sample preparation is key to usefulness of data obtained!!!

32. Fluorescent Sperm Viability Assay • Sybr14 / PI • Green = viable • Red = dead • Adaptable to Microscopy or Flow cytometry

33. Morphology • Variety of sample preparations and methods for evaluation • Stain vs. no stain • Wet mount vs. dried or fixed • Magnification of observation • Computer systems require training samples

34. Morphology Classifications • Scheme A • Primary – Head abnormalities • Secondary – Tail abnormalities • Scheme B • Primary – Head abnormalities • Secondary – Tail abnormalities • Tertiary – Cytoplasmic droplets

37. Ejaculate Characteristics Species Ejaculate Sperm Total Sperm % % Volume Conc. (x109) Motile Normal (x109/ml) /Ejaculate Bull 8.0 1.5 12 75 95 Ram 1.0 3.0 3 95 95 Boar 200 .25 50 70 90 Stallion 80 .15 12 70 40-90 Man 2-6 .15 .9 65 30-70

38. Impact of Heat on Spermatogenesis L Enwall, J. Schindler, K Gibbs, K Yagoda and J. Parrish

39. Raises Scrotal Temperature 1.9°C (3.4°F)

41. Normalization of Data • (Sample value) – (Pre-insulation value) • Normalized for each boar • On day 0, all graphed data should be 0 • Data • Primary Abnormalities • Secondary Abnormalities • Tertiary Abnormalities • Motile Sperm

46. Effect of Scrotal Insulation on Primary Abnormalities Development During Boar Spermatogenesis

47. Effect of Scrotal Insulation on Secondary Abnormalities Development During Boar Spermatogenesis

48. Effect of Scrotal Insulation on Tertiary Abnormalities Development During Boar Spermatogenesis

49. Effect of Scrotal Insulation on Development of Motility Potential During Boar Spermatogenesis

50. Impact of Scrotal Insulation on Spermatogenesis in the Boar • Pachytene Primary Spermatocyte • Morphology • Motility • Total sperm output