THE EFFECTS OF LIGATION OF CAUDA EPIDIDYMIDIS ON THE DOG TESTIS

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1 Copyright 1974 The American Fertility Society FERTILITY AND STERILITY Vol. 25, No.3, March, 1974 Printed in U.S.A. THE EFFECTS OF LIGATION OF CAUDA EPIDIDYMIDIS ON THE DOG TESTIS A. M. VARE, M.B.B.S., M.Sc. (MED.), M.A.M.S., AND P. C. BANSAL, M.S., F.R.C.S., M.A.M.S. Department of Anatomy, Medical College Aurangabad (M.S.), India Researchers studying the effects of ligation of the epididymis at different levels and ductuli efferenti on animal testes are not in agreement. It has been reported that the ligation of efferent ducts or caput epididymidis produces substantial testicular damage. I - lo On the other hand, obstruction of cauda epididymidis does not affect the testes.u Baillie 7 and Paufler and Foote I2 have reported that after ligation of epididymis at different levels (caput or corpus or cauda), the seminiferous tubules undergo two phases in testicular response; first, the seminiferous tubules dilate, change, then, completely recover. Also, the closer the ligature of the epididymis to the testis, the more damage to the testis. At present, vasectomy is preferably performed by scrotal incisions; therefore, the cauda epididymidis may be ligated instead of the vas deferens. The purpose of this study was to ligate canine cauda epididymidis and compare our findings with those of earlier researchers. MATERIALS AND METHODS Twelve adult dogs weighing 20 to 25 kg were used in this study. Under ether anesthesia, a short incision was made on the left side of scrotum and tunica vaginalis, exposing the testis. The left testis and the epididymis was delivered and the left cauda epididymidis was ligated with a Received August 9, fine suture material. Care was taken not to ligate a blood vessel. The tunica vaginalis and the scrotum were sutured separately. The right testis remained in situ as a control. Two dogs were killed every 15 days for 3 months and the gonads and epididymis were removed. The size, consistency, and external appearance of the testes and epididymides were noted. The testes were fixed in Bouin's fluid, sectioned at 7 }-t, and stained with hematoxylin, eosin, periodic Acid-Schiff Reaction, and van Gieson's solution. The results of histologic examination of ligated testes were compared with the control testes removed from the same animal. A normal seminiferous tubule from a control testis is shown in Figure 1. RESULTS 15 Days After Ligation. Macroscopic examination of the ligated testes revealed that they were larger and more turgid than the control sides. Microscopic examination of experimental gonads showed that there was an increase in the lumen and diameter of most of the seminiferous tubules. The lumen of the tubules contained shed spermatozoa and fluid (Fig. 2). The germinal epithelium was normal except that there were a few sperm attached to it. The intertubular tissue showed no changes. 1 Month After Ligation. The ligated

2 VO'I. 25, NO'. 3 EFFECTS OF CAUDA EPIDIDYMIDIS LIGATION FIG. 1. Normal seminiferous tubule lined by seven or eight cell layers and showing various stages of spermatogenesis. Note the many sperm attached to the germinal epithelium. (x60) FIG. 2. Seminiferous tubule 15 days after ligation. The tubule is lined by seven or eight cell layers and different stages of spermatogenesis are seen, but only a few sperm are attached to germinal epithelium. The lumen of the tubule is dilated and contains shed sperm. (x60) 257

3 258 VARE AND BANSAL March 1974 FIG. 3. Seminiferous tubules 1 month after ligation. The tubule (arrow) is lined by six or seven cell layers. The outline of most of the cells of the germinal epithelium is indistinct and their nuclei show pyknotic changes. The intertubular stroma exhibit inflammatory cells. The Leydig cells are unchanged. (x60) FIG. 4. Section of testis 1 Yz months after ligation. The tubules are lined by three or four cell layers. The degenerative changes in the cells of the germinal epithelium are advanced. Most of the tubules appear smaller than the control. The intertubular stroma shows an increase in fibrous tissue and macrophages. (x60)

4 Vol. 25, No.3 EFFECTS OF CAUDA EPIDIDYMIDIS LIGATION 259 FIG. 5. Section of testis 2 months after ligation. The tubules show striking degenerative changes. They are lined by one or two cell layers and they are much smaller than the controls. The intertubular stroma exhibit a marked increase in fibrous tissue and macrophages. (x60) testes were smaller and softer than those of control testes. Histologic examination of ligated testes showed germinal epithelium degeneration; it decreased from seven or eight cell layers to five or six. The degenerating cells were hazy and their nuclei were pyknotic (Fig. 3). The Leydig cells were normal, but the intertubular stroma contained a few inflammatory cells. 1 % Months After Ligation. The experimental testes were softer and smaller than the controls. The degenerative changes were more advanced in the seminiferous tubules and most of the tubules were smaller than the controls (Fig. 4). The germinal epithelium was reduced to three or four cell layers. The intertubular stroma showed an increase in the collagen fibers, lymphocytes, and macrophages. 2 Months After Ligation. The size of seminiferous tubules of ligated testes was reduced; therefore, the number of tubules per field was greater than in the control testes (Fig. 5). The seminiferous tubules presented marked degenerative changes and they were lined by one or two cell layers. Some tubules were replaced by fibrous tissue (Fig. 5). The intertubular stroma showed much more increase In collagen fibers and inflammatory cells. 2% and 3 Months After Ligation. The seminiferous tubules and intertubular stroma appeared similar to those of 2 months after ligation. DISCUSSION Testis size was found to increase during the first 15 days after ligation of cauda epididymidis and then decrease progressively for 1 to 3 months. The initial increase in size and turgidity is probably due to an increase in pressure within the tubules. The testis decreases in size and becomes soft 1 to 3 months after

5 260 V ARE AND BANSAL March 1974 ligation because of the degenerative changes in the seminiferous tubules and the gradual reduction in intra tubular pressure. The atrophic and degenerative changes of the seminiferous tubules probably result from blood stasis which, in turn, is caused by the resistance of fibrous tunica albuginea to increase in internal pressure. Blood stasis may limit oxygen and therefore injure the sensitive germinal epithelium. Baillie 7 and Paufler and Foote'2 also observed initial lumen increase and seminiferous tubule changes but these changes were temporary, and the tubules eventually became more or less normal. SUMMARY Testis size increased and the testes became more turgid during the first 15 days after ligation of the cauda epididymidis of 12 dogs. However, after 15 days, the size of testes gradually decreased and the testes became soft. Microscopic examination of testes during the first 15 days revealed that there was an increase in lumen and diameter of most of the seminiferous tubules, but the germinal epithelium and intertubular tissue did not show any changes. One to three months after ligation, the seminiferous tubules underwent degenerative changes. The tubules became smaller and were lined by one or two cell layers. Some tubules were replaced by fibrous tissue. The intertubular stroma showed an increase in collagen fibers and macrophages, but the Leydig cells were unchanged. REFERENCES 1. White EW: Duration of fertility and histological changes in reproductive organs after ligation of vasa efferentia in rat. Proc R Soc Lond [BioI] 113:544, van Wagenen G: Degeneration of germinal epithelium in the testis of the rat as a result of efferent duct ligation. Anat Rec 27: 189, van Wagenen G: Changes in the testis of rat following ligation of ductuli efferenti. Anat Rec 29:399, Harrison RG: The effect of ligation of vasa efferentia on the rat testis. Proc Soc Study Fertil 5:97, Harrison RG, MacMillan EW: The effect of high epididymal obstruction upon the Leydig cell volume of rat testis. J Endocrinol 11: 89, Baillie AH: The biology of Leydig cell: histochemical and histological changes following high epididymal obstruction. J Endocrinol 20:339, Baillie AH: Observations comparing the effects of epididymal obstruction at various levels on the mouse testis with those of ischaemia. J Anat 96:335, Glover TD: The response of rabbit spermatozoa to artificial cryptorchidism and ligation of epididymis. J Endocrinol 23:317, Barack BM: Transport of spermatozoa from seminiferous tubules to epididymis in the mouse: a histological and quantitative study. J Reprod Fertil 16:35, Igboeli G, Foote RH: Maturation and aging changes in rabbit spermatozoa isolated by ligatures at different levels of epididymis. Fertil Steril 20: 506, MacMillan EW: The effect of interruption of vasal and inferior epididymal arteries on cauda epididymis and testis. Proc Soc Study Fertil 5: 12, Paufler SK, Foote RH: Spermatogenesis III rabbit following ligation of epididymis at different levels. Anat Rec 164:339, 1969

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