FERTILITY AND STERILITY Copyright c 1975 The American Fertility Society
Vol. 26, No. 1, January 1975 Printed in U.S.A.
MICROPUNCTURE AND MICROANALYTIC STUDIES OF THE EFFECT OF VASECTOMY ON THE RAT TESTIS AND EPIDIDYMIS* STUARTS. HOWARDS, M.D., SANDRA JESSEE, B.S.,
AND
ANNE JOHNSON, M.S.
Male Reproductive Micropuncture Laboratory, Department of Urology, Charlottesville, Virginia 22901
In 1924, Moore and Quick wrote, "Vasectomy and its effects upon the mammalian testis have been discussed for more than half a century." 1 During the ensuing four decades, interest in the study of vasectomy waned in this country. However, investigation of the effects of vasoligation has again intensified, perhaps because millions of men have undergone bilateral partial vasectomy to achieve permanent infertility. There is general agreement that vasectomy is a relatively safe, reliable, and inexpensive means of contraception. Nevertheless, several studies have raised the possibility of untoward effects of the procedure. 2 .a Although these reports are not well documented, publicity has created a demand for further investigation of undesirable effects of vasectomy. Our work was initiated to study the effects of occlusion of the vas deferens on the physiology of the testis and epididymis of the rat by means of micropuncture and microanalytic techniques.
thetized with intraperitoneally injected sodium pentobarbital (Nembutal) in a dose of 40 mg/kg body weight, and the left vas deferens was exposed through a 1.5 em vertical incision. Initially, the procedure for vasectomy involved dissection of the vas deferens from the surrounding sheath of fat. Care was taken not to occlude the major deferential blood vessels. Interruption of the vas was then accomplished by removing 5-mm section from between two 6-0 silk ties. Since this method invariably led to formation of a spermatic granuloma at the proximal cut end of the vas, several modifications of the initial procedure were tried. Silk, nylon, and chromic catgut sutures varying in size from 4-0 to 6-0, metallic clips, and electrocautery were all used, both with and without resection of a segment of the vas deferens. In addition, the distance of the partial vasectomy from the cauda epididymidis was varied. Unfortunately, none of these modifications decreased the incidence of spermatic granulomas, and only when the vas was MATERIALS AND METHODS left in its surrounding fat sheath, regardless of the method of interruption, was Male Sprague-Dawley rats, weighing the incidence reduced from 100% to apfrom 300 to 350 gm (Flow Research Aniproximately 85%. After vasectomy, the mals, Dublin, Va), were allowed to acanimals were divided into two groups, climate to their new surroundings for at the first to be studied after one week and least one week. the second after one month. For vasectomy, the animals were anesAt the appropriate interval following *Supported in part by NIH-NICHD contract vasectomy, the rats were anesthetized 72-2770 and the James McCahill Urology Research with 100 mg/kg body weight of intraFund. peritoneal sodium 5-ethyl-5(1-methyl 20
Vol. 26, No.1
21
MICROPUNCTURE STUDIES: II
FIG. 1: Spennatic granuloma at the ligation site on the vas deferens of a rat, one week postvasectomy. The granuloma is the small white oval structure along the vas deferens below the testis.
propyl)-2-thiobarbiturate (lnactin). The trachea was intubated, and the animal was placed on a constant-heat, small animal table and prepared for micropuncture by the method described previously.4 If a spermatic granuloma was present, the animal was placed in the granuloma subgroup (G); if not, it was assigned to the vasectomy without granuloma category (NG). The contents of each granuloma were aspirated with a micropipette, placed on a glass slide, stained with rose bengal, and examined microscopically to confirm the presence of spermatozoa. Microsamples of from 75 to 200 nl (1 nl = 10-6 ml) were then obtained in vivo from the left vasectomized testis, caput epididymidis, and cauda epididymidis. The micropuncture techniques and the puncture sites have been. previously described.4 Next, the right (control) testis
and epididymis were exposed, and samples from sites comparable to those on the left side were taken as one type of control. Following the collections, each defatted testis and epididymis was weighed on a Mettler analytical balance. The microsamples were then prepared for the determination of spermatocrit or for morphologic study as previously described. 4 The percentage of abnormal sperm on eaOC.OO)ll:> ll:>~ll:>'~~o ll:>~ll:>ll:>t-
cicicicici ..-1~0)..-lt:'l
,.....;,.....;,.....;,.....;,.....
tl "' ' O)o t- •
......· 0
C'IO)' t"'
t".l
5.0±0.6e
~
;::s ~
1;:1
~
.....
~
~
MICROPUNCTURE STUDIES: II
Vol. 26, No.1
ever, after vasectomy without granuloma on both the left and right sides, there were more abnormal spermatozoa in the caput than in the cauda
Vol. 26, No. 1, January 1975 Printed in U.S.A.
MICROPUNCTURE AND MICROANALYTIC STUDIES OF THE EFFECT OF VASECTOMY ON THE RAT TESTIS AND EPIDIDYMIS* STUARTS. HOWARDS, M.D., SANDRA JESSEE, B.S.,
AND
ANNE JOHNSON, M.S.
Male Reproductive Micropuncture Laboratory, Department of Urology, Charlottesville, Virginia 22901
In 1924, Moore and Quick wrote, "Vasectomy and its effects upon the mammalian testis have been discussed for more than half a century." 1 During the ensuing four decades, interest in the study of vasectomy waned in this country. However, investigation of the effects of vasoligation has again intensified, perhaps because millions of men have undergone bilateral partial vasectomy to achieve permanent infertility. There is general agreement that vasectomy is a relatively safe, reliable, and inexpensive means of contraception. Nevertheless, several studies have raised the possibility of untoward effects of the procedure. 2 .a Although these reports are not well documented, publicity has created a demand for further investigation of undesirable effects of vasectomy. Our work was initiated to study the effects of occlusion of the vas deferens on the physiology of the testis and epididymis of the rat by means of micropuncture and microanalytic techniques.
thetized with intraperitoneally injected sodium pentobarbital (Nembutal) in a dose of 40 mg/kg body weight, and the left vas deferens was exposed through a 1.5 em vertical incision. Initially, the procedure for vasectomy involved dissection of the vas deferens from the surrounding sheath of fat. Care was taken not to occlude the major deferential blood vessels. Interruption of the vas was then accomplished by removing 5-mm section from between two 6-0 silk ties. Since this method invariably led to formation of a spermatic granuloma at the proximal cut end of the vas, several modifications of the initial procedure were tried. Silk, nylon, and chromic catgut sutures varying in size from 4-0 to 6-0, metallic clips, and electrocautery were all used, both with and without resection of a segment of the vas deferens. In addition, the distance of the partial vasectomy from the cauda epididymidis was varied. Unfortunately, none of these modifications decreased the incidence of spermatic granulomas, and only when the vas was MATERIALS AND METHODS left in its surrounding fat sheath, regardless of the method of interruption, was Male Sprague-Dawley rats, weighing the incidence reduced from 100% to apfrom 300 to 350 gm (Flow Research Aniproximately 85%. After vasectomy, the mals, Dublin, Va), were allowed to acanimals were divided into two groups, climate to their new surroundings for at the first to be studied after one week and least one week. the second after one month. For vasectomy, the animals were anesAt the appropriate interval following *Supported in part by NIH-NICHD contract vasectomy, the rats were anesthetized 72-2770 and the James McCahill Urology Research with 100 mg/kg body weight of intraFund. peritoneal sodium 5-ethyl-5(1-methyl 20
Vol. 26, No.1
21
MICROPUNCTURE STUDIES: II
FIG. 1: Spennatic granuloma at the ligation site on the vas deferens of a rat, one week postvasectomy. The granuloma is the small white oval structure along the vas deferens below the testis.
propyl)-2-thiobarbiturate (lnactin). The trachea was intubated, and the animal was placed on a constant-heat, small animal table and prepared for micropuncture by the method described previously.4 If a spermatic granuloma was present, the animal was placed in the granuloma subgroup (G); if not, it was assigned to the vasectomy without granuloma category (NG). The contents of each granuloma were aspirated with a micropipette, placed on a glass slide, stained with rose bengal, and examined microscopically to confirm the presence of spermatozoa. Microsamples of from 75 to 200 nl (1 nl = 10-6 ml) were then obtained in vivo from the left vasectomized testis, caput epididymidis, and cauda epididymidis. The micropuncture techniques and the puncture sites have been. previously described.4 Next, the right (control) testis
and epididymis were exposed, and samples from sites comparable to those on the left side were taken as one type of control. Following the collections, each defatted testis and epididymis was weighed on a Mettler analytical balance. The microsamples were then prepared for the determination of spermatocrit or for morphologic study as previously described. 4 The percentage of abnormal sperm on eaOC.OO)ll:> ll:>~ll:>'~~o ll:>~ll:>ll:>t-
cicicicici ..-1~0)..-lt:'l
,.....;,.....;,.....;,.....;,.....
tl "' ' O)o t- •
......· 0
C'IO)' t"'
t".l
5.0±0.6e
~
;::s ~
1;:1
~
.....
~
~
MICROPUNCTURE STUDIES: II
Vol. 26, No.1
ever, after vasectomy without granuloma on both the left and right sides, there were more abnormal spermatozoa in the caput than in the cauda
