James K. Graham, PhDProfessor Office: 247 Phyiology Building Member Education Link to a PubMed listing of Dr. Graham's
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Teaching Activities
I teach male reproductive physiology to veterinary students and graduate students. I also teach endocrinology to undergraduate (BS 430) and graduate students and offer a course in Human Health and Disease (BSCC 120) to undergraduate students.
Research Interests -- Male Reproductive Physiology
My research focuses on gamete physiology and cryobiology, with an emphasis on membrane biochemistry and function. Membranes in spermatozoa are modified extensively during transit through the epididymis and the female reproductive tract prior to fertilization, or during in vitro manipulations such as cryopreservation.I am interested in understanding the membrane modifications that occur during these processes, i.e., what changes occur, when they occur, and how they affect cell function. Using liposome technologies, membranes of sperm can be changed in a predetermined way with a subsequent alteration of cell function. Changes in sperm membranes can be quantified using flow cytometry. Present studies investigate mechanisms by which liposomes interact with cells to: 1) induce an acrosome reaction in sperm and allow their subsequent penetration into eggs in vitro; and 2) modify membranes to permit cell survival during cryopreservation.Cryopreservation of cells is poorly understood, and cryopreserved spermatozoa behave differently than unfrozen cells. Only for cattle is fertility of frozen-thawed spermatozoa satisfactory. Research in my laboratory is directed toward understanding some of the basic principles of freezing cells. These include changes in the membranes and cytoplasm induced by cryoprotectants and low temperatures, and the interactions between frozen/thawed spermatozoa and the epithelium of female tract and/or the egg. These studies involve both in vivo systems and in vitro culture systems. Using the chicken as a model, these studies should lead to improved methods for the preservation of cells in general, and spermatozoa specifically.
For additional information on current projects, consult the page on Gamete Biology and Andrology.
Representative Publications
Graham JK, Kunze E, Hammerstedt RH. 1990. Analysis of sperm cell viability, acrosomal integrity, and mitochondrial function using flow cytometry. Biol Reprod 43:55-64.
Graham JK, Nolan JP, Hammerstedt RH. 1991. Effect of dilauroylphosphatidylcholine liposomes on motility, induction of the acrosome reaction and subsequent egg penetration of epididymal ram sperm. Biol Reprod 44:1092-1099.
Parks JE, Graham JK. 1992. Effects of cryopreservation on sperm membranes. Theriogenology 38:210-222.
Phillips JJ, Bramwell RK, Graham JK. 1996. Effect of methyl cellulose on the cryopreservation of rooster sperm. Poultry Sci 75:915-923.
Truelson SL, Graham JK, Mortimer RG, Field TG. 1996. The in vitro penetration into bovine oocytes and zona-free hamster oocytes by spermatozoa treated with liposomes. J Dairy Sci 79:991-999.
Dalimata AM, Graham JK. 1997. Cryopreservation of rabbit spermatozoa using acetamide in combination with trehalose and methyl cellulose. Theriogenology 48:831-841.
Denniston DJ, Graham JK, Squires EL, Brinsko SP. 1997. The effect of liposomes composed of phosphatidylserine and cholesterol on fertility rates using frozen-thawed equine spermatozoa. J Equine Vet Sci 17:675-676.
Parrish JJ, Susko-Parrish JL, Graham JK. 1999. In vitro capacitation of bovine spermatozoa: Role of intracellular calcium. Theriogenology 51:461-472.
Denniston DJ, Squires EL, Bruemmer JE, Brinsko SP, McCue PM, Graham JK. 2000. Effect of antioxidants on the motility and viability of cooled stallion spermatozoa. J Reprod Fert, Suppl 56:121-126.
Brinsko SP, VanWagner GS, Graham JK, Squires EL. 2000. Motility, morphology and triple stain analysis of fresh, cooled and frozen-thawed stallion spermatozoa. J Reprod Fert, Suppl 56:111-120.
Dewit M, Marley WS, Graham JK. 2000. Fertilizing potential of mouse spermatozoa cryopreserved in a medium containing whole eggs. Cryobiology 40:36-45.
Salazar P, Graham JK, Parrish JJ, Susko-Parrish J, Squires EL. 2000. Indirect determination of stallion sperm capacitation based on esterase release from spermatozoa challenged with lysophosphatidylcholine. J Reprod Fert, Suppl 56:407-414.
Crockett EC, Graham JK, Bruemmer JE, Squires EL. 2001. Effect of cooling of equine spermatozoa before freezing on post-thaw motility: Preliminary results. Theriogenology 55:793-803.
Landim-Alvarnega FC, Alvarenga MA, Seidel GE Jr. 2001. Penetration of zona-free hamster, bovine and equine oocytes by stallion and bull spermatozoa pretreated with equine follicular fluid, dilauroylphosphatidylcholine or calcium ionophore A23187. Theriogenology 56:937-953.
Graham JK. 2001. Assessment of sperm quality: A flow cytometric approach. Anim Reprod Sci 68:239-248.
Bruemmer JE, Coy RC, Squires EL, Graham JK. 2002. Effect of pyruvate on the function of stallion spermatozoa stored for up to 48 hours. J Anim Sci 80:12-18.
Navratil AM, Bliss SP, Berghorn KA, Haughian JM, Farmarie TA, Graham JK, Clay CM, Roberson MS. 2003. Constitutive localization of the gonadotropin-releasing hormone (GnRH) receptor to low-density membrane microdomains is necessary for GnRH signaling to ERK. J Biol Chem 278: 31593-31602.
Purdy PH, Graham JK. 2004. Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm. Cryobiology 48:36-45.
Purdy PH, Graham JK. 2004. Effect of adding cholesterol to bull sperm membranes on sperm capacitation, the acrosome reaction and fertility. Biol Reprod 71:522-527.
Squires EL, Keith SL, Graham JK. 2004. Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology 62:1056-1065.
Landim-Alvarenga FC, Graham JK, Alvarenga MA, Squires EL. 2004. Calcium influx into equine and bovine spermatozoa during in vitro capacitation. Anim Reprod 1:96-105.
Moore AI, Squires EL, Graham JK. 2005. The effect of seminal plasma on the cryopreservation of equine spermatozoa. Theriogenology 63:2372-2381.
Kirk ES, Squires EL, Graham JK. 2005. Comparison of in vitro laboratory analyses with the fertility of cryopreserved stallion spermatozoa. Theriogenology 64:1422-1439.
Purdy PH, Fox MH, Graham JK. 2005. The fluidity of Chinese hamster ovary cell and bull sperm membranes after cholesterol addition. Cryobiology 51:102-112.
Graham JK, Mocé E. 2005. Fertility evaluation of frozen/thawed semen. Theriogenology 64:492-504.
Moore AI, Squires EL, Graham JK. 2005. Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology 51:241-249.
Mocé E, Graham JK. 2006. Cholesterol-loaded cyclodextrins added to fresh bull ejaculates improve sperm cryosurvival. J Anim Sci 84:826-833.
