Characterization of sperm protein at 22 KDA (SP22) in the stallion reproductive tract
by Miller, Lynda Mae Jean, Ph.D., UNIVERSITY OF KENTUCKY, 2010, 156 pages; 3498026

Abstract:

SP22 has been described and proven to be related to fertility in the rat, but no information is available in horses. This dissertation was completed to characterize SP22 in the reproductive tract of the stallion. Five hypotheses were tested: (1) SP22 is present on stallion spermatozoa; (2) SP22 is expressed in testicular and epididymal tissues in stallions; (3) Environmental damage of equine spermatozoa results in alteration of the expression and localization pattern of SP22 on spermatozoa; (4) Insult to the testes affects the expression and localization of SP22 on ejaculated and caudal epididymal spermatozoa; and (5) Insult to the testes affects the expression of SP22 in the reproductive tract of stallions.

To determine if SP22 was present on stallion spermatozoa, immunocytochemistry (ICC) was performed and the distribution pattern(s) of SP22 was determined. The protein was determined to be present. To test if SP22 was expressed in the reproductive tract of stallions, tissues were collected during routine castrations. Protein and RNA were extracted and polymerase chain reaction (PCR) and Northern Blot analysis were performed. Western Blot analysis confirmed the presence and size of the SP22 protein. It was determined that SP22 protein and RNA are present in the stallion's reproductive tract. To determine if cooling and freeze/thawing of spermatozoa affected the expression and localization of SP22 on spermatozoa ICC was performed on ejaculates of stallions prior to and following cooling and a routine freezing protocol. It was determined that the expression was altered by environmental damage. Hypotheses four and five determined that the localization of SP22 and expression of SP22 protein and RNA on the spermatozoa was, in fact, altered following testicular insult.

When the results of all five experiments are considered in concert, it was determined that SP22 is present on spermatozoa and in testicular and epididymal tissues. Furthermore, environmental and physiologic damage to spermatozoa alters the expression of SP22.

KEYWORDS: Sperm Protein, SP22, Plasma Membrane Proteins, Equine Spermatozoa, Stallion Reproduction

 
AdviserMats H.T. Troedsson
SchoolUNIVERSITY OF KENTUCKY
SourceDAI/B 73-06, p. , Apr 2012
Source TypeDissertation
SubjectsCellular biology; Veterinary medicine
Publication Number3498026
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