This proposal focusses on definition of the critical differences underlying fertility and infertility after vasovasostomy, and the analysis of immunodominant post-vasectomy sperm autoantigens using monoclonal antibodies and gene cloning. In most of the experiments, rats will receive a bilateral vasectomy and a later vasovasostomy, a vasectomy alone, or sham operations. Among our previous observations in this model were testicular and epididymal alterations, correlations between antisperm antibodies and reproductive tract changes, and definition of immunodominant sperm autoantigens post-vasectomy and vasovasostomy. Some vasectomy-induced responses were not reversed by a subsequent vasovasostomy.
The first aim of the proposed work is to determine the basis for differences in fertility after vasovasostomy, utilizing Lewis and Fischer strains of rats, which differ dramatically in fertility after vasovasostomy but share histocompatibility loci.
The second aim i s to identify the types of leukocytes that appear within the reproductive tract in reaction to vasectomy and vasovasostomy using immunohistochemical studies.
The third aim i s to capture rat monoclonal autoantibodies to dominant post-vasectomy sperm autoantigens by constructing rat x rat hybridomas.
The fourth aim i s to clone and sequence cDNAs for the dominant post-vasectomy sperm autoantigens, to characterize the autoantigens, and the utilize the nucleotide and protein probes thereby made available for further studies in rats and humans. Methods include light and electron microscopy, immunohistochemistry, determination of serum antisperm autoantigens, micropuncture for measurement of intraluminal hydrostatic pressure, and determination of the resistance of the vas deferens to fluid flow. Standard methods of molecular biology will be employed to clone cDNAs and analyze recombinant sperm autoantigens.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD018825-08
Application #
3315985
Study Section
Reproductive Biology Study Section (REB)
Project Start
1984-09-01
Project End
1995-04-30
Budget Start
1992-05-01
Budget End
1993-04-30
Support Year
8
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Virginia
Department
Type
Schools of Medicine
DUNS #
001910777
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Flickinger, C J; Bush, L A; Howards, S S et al. (1997) Distribution of leukocytes in the epithelium and interstitium of four regions of the Lewis rat epididymis. Anat Rec 248:380-90
Flickinger, C J; Howards, S S; Baran, M L et al. (1997) Appearance of 'natural' antisperm autoantibodies after sexual maturation of normal Lewis rats. J Reprod Immunol 33:127-45
Flickinger, C J; Baran, M L; Howards, S S et al. (1996) Sperm autoantigens recognized by autoantibodies in developing rats following prepubertal obstruction of the vas deferens. J Androl 17:433-42
Flickinger, C J; Herr, J C; Baran, M L et al. (1995) Temporal appearance of antisperm antibodies during sexual maturation of rats after obstruction of the vas deferens. J Androl 16:75-9
Flickinger, C J; Herr, J C; Baran, M L et al. (1995) Testicular development and the formation of spermatic granulomas of the epididymis after obstruction of the vas deferens in immature rats. J Urol 154:1539-44
Flickinger, C J; Howards, S S; Herr, J C (1995) Effects of vasectomy on the epididymis. Microsc Res Tech 30:82-100
Flickinger, C J; Howards, S S; Bush, L A et al. (1995) Antisperm autoantibody responses to vasectomy and vasovasostomy in Fischer and Lewis rats. J Reprod Immunol 28:137-57
Flickinger, C J; Harris, M; Herr, J C et al. (1994) Early antibody response following vasectomy is related to fertility after vasovasostomy in glucocorticoid-treated and untreated Lewis rats. J Urol 151:791-6
Flickinger, C J; Howards, S S; Bush, L A et al. (1994) Temporal recognition of sperm autoantigens by IgM and IgG autoantibodies after vasectomy and vasovasostomy. J Reprod Immunol 27:135-50
Flickinger, C J; Herr, J C; Sisak, J R et al. (1993) Ultrastructure of epididymal interstitial reactions following vasectomy and vasovasostomy. Anat Rec 235:61-73

Showing the most recent 10 out of 25 publications