Effects of Ablation of the Submaxillary Gland in Guinea Pigs IV. Cause of deterioration of the tubules in the testes

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1 Effects of Ablation of the Submaxillary Gland in Guinea Pigs IV. Cause of deterioration of the tubules in the testes Kazuo Suzuki Received August 1, 1960 Shakujii Institute, Tokyo Medical College, Tokyo, Japan Introduction The atrophic change of germ cells in the testis after the submaxil lariectomy in the guinea pigs has been reported by the writer in the previous paper (1957). Morphologically atrophic change of germ cells in the testis after the submaxillariectomy closely resembled the deterioration of testicular tubules in the hypohysectomized animals. Therefore, the cause of deteriora tion of germ cells in the testis after the submaxillariectomy was thought to be a result following the change in the androgen that has arisen from the submaxillariectomy. However, as stated in the previous paper, the lack of correlation between the conditions of testis and seminal vesicle was observed. The above reported facts raised a question of hormonal consideration about the deterioration of germ cells in the testis by the submaxillariectomy. A new interpretation as to the deterioration of tubules in the testis after the submaxillariectomy will be stated in this paper. Material and methods Guinea pigs used in the present work were the progeny of the inbred stock maintained in our laboratory. The testes, ranging from 3 days to 180 days after submaxillariectomy, were fixed with Bouin's fluid. The sections were cut in 5-8 micra in thickness, and stained separately with Heidenhain's iron haematoxylin and Delafield's haematoxylin eosin. observations The histological examination of testis after the submaxillariectomy re vealed an extensive degenerative change in the germinal epithelium. Fig. 1 shows the lobule found in testis three days after operation. A great amount of fluid was stained deep red pouring into the lobular cavity, whereas, the germ cells of intact are seen in the germinal epithelium. Figs. 2 and 3 show progressive deterioration of the germinal epithelium. The germ cells are reduced in numbers, and space devoid of intercellular

2 Figs , section through the lobule of testis 3 days after the submaxillariectomy. It is noted that a great amount of water fluid flowed into the lobular cavity, and germ cells were intact in the lobular epithelium. 2, beginning of the deterioration of lobular epithelium, and masses of intercellular substances containing the sperms were broken loose from the lobular epithelium. 3, section through the same testis of Fig. 2, showing the progressive deterioration of lobular epithelium. 4, section through the lobule of testis 14 days after operation, showing degenerating lobule. Masses of intercellular substances and germ cells lie in the lobular cavity. Scattered germ cells in the lobular epithelium devoid of intercellular substances suffered no damage. 5, a lobule after the deterioration, degenerating spermatocyte cells and fibrin-like pattern in the site of lobular epithelium, and masses of intercellular substances, degenerating germ cells and sperms in the lobular cavity. 6-7, sections through the lobules of testis 13 days after operation, showing the liberation of germ cells embedded in the masses of intercellular substances like the cyst. 8, section through the testis 30 days after operation showing the empty lobules, fibrin-like network in the site of lobular epithelium. Sertoli b cells and spermatogonial cells lining on the inner order of the lnhule.

3 1961 Effects of Ablation of the Submaxillary Gland in Guinea Pigs IV 477 substances is seen in the germinal epithelium. The sperms are embedded in the massed intercellular substances and seem to flow into the lobular cavity. Fig. 4 shows the lobule of the testis on the 7 days after operation, the diminution of intercellular substances and scattered germ cells can be seen in the germinal epithelium, but it is a noticeable aspect that most germ cells have not yet suffered the impairment of intercellular substances. The massed young cells and spermatazoa, and mass of intercellular substances are seen in the lobular cavity. Fig. 5 shows the lobule after the deterioration. Most germ cells are broken loose from the lobular epithelium and the fibrin-like pattern, which is clotted intercellular substances, and a few spermatocyte cells remained in the germinal epithelium. The degenerating germ cells, spermatozoa and massed intercellular substances lie scattered in the lobular cavity. Figs. 6 and 7 show the lobules of testis 14 days after the operation, a number of germ cells are embedded in the massed intercellular substances like the cyst, which will be seen broken loose from the germinal epithelium. Fig. 8 shows a part of the adjoining lobules after the deterioration, and the lobules are poorly populated with germ cells. The spermatogonial cells and sertoli cells remained adjacent to the basement membrance and a fibrillar network is seen in the site of germinal epithelium. Discussion The testis in the submaxillariectomized guinea pigs caused atrophic changes of germinal epithelium. This process of change was found in the testis 3 days after the operation. The process of the deterioration in the germinal epithelium could be observed in the testis after the submaxillariectomy, namely: 1) the watery fl uid flowed into the lobular cavity as seen in Fig. 1, 2) the liberation of the mass of intercellular substances, germ cells contained and spermatozoa from the germinal epithelium (Figs. 2, 3, 6 and 7), and 3) the fibrin-like network appeared in the site of germinal epithelium after the deterioration of tubules (Figs. 5 and 8). It seemed logical to assume that a change to excrete the watery fluid, and to transmute into the jelly-like mass may have appeared in the coagula tion of intercellular substances. From the above stated fact the cause of deterioration of tubules in the testis after operation is thought to be the result of clotting in the intercel lular substances that has arisen from the submaxillariectomy. It is a noteworthy fact that germ cells in the germinal epithelium which is devoid of intercellular substances, and germ cells embedded in the massed intercellular substances, seemed to suffer no damage. Therefore, it will be seen that deterioration of intercellular substances Cytologia

4 478 K. Suzuki Cytologia 26 apparently occurred earlier than the damage of germ cells incurred by the submaxillariectomy. Judging from the facts stated above, it may be surmised that atrophic changes of the germ cells in the testis are due to the result of deterioration of intercellular substances by the submaxillariectomy. Various authors reported on the morphological aspects that the deterio ration of tubules in the testis after the submaxillariectomy in the guinea pigs closely resembles the disorder of the spermatogenetic process in the testis of hypohysectomized animals. Thus, in the previous paper (1957), the present writer reported that the cause of atrophic change of germ cells in the testis could be conjectured as a result after the change in the androgen that has arisen from the sub maxillariectomy. However, as reported in the same paper (1957), the lack of correlation between the conditions of testis and seminal vesicle was observed in the specimen after operation, namely, the germ cells and sperms suffered damage, whereas the seminal vesicle exhibited normal development. Such hormonal consideration about the atrophic change of germ cells in the testis after the submaxillariectomy needs further exploration. Judging from the results of the present observation, the atrophic change of germ cells in the testis of submaxillariectomized guinea pigs may be due to the results of clotting in the intercellular substances which has arisen from the submaxillariectomy. In the experimental studies on the female guinea pigs by the submaxil lariectomy, the writer (1961) has reported that secretion of the submaxillary gland presumably contains the anti-coagulate enzyme for the follicular fluid and tissue fluid in the ovarian stroma. Similarly, it may be surmised that secretions of the submaxillary gland in the male guinea pigs contains the anti-coagulate enzyme for the inter cellular substances in the germinal epithelium. One of the surprising things is that the deterioration of tubules in the abdominal testis showed close similarity to the morphological aspects that atrophic changes of tubules were found in the testis of the submaxillari ectomized guinea pigs. The writer's interpretation with regard to the cause of progressive deterioration of tubules in the abdominal testis of the guinea pigs will be outlined below. The peritubular fibrosis in the abdominal testis is thought to be the results of clotting in the intercellular substances that has arisen from elevated temperature, and an arrested spermatogenetic progress may be due to the result of deterioration of intercellular substances in the germinal epithelium.

5 1961 Effects of Ablation of the Submaxillary Gland in Guinea Pigs IV 479 Summary 1. The nature of submaxillary glands and the cause of deterioration of the tubules in the testis of guinea pigs were studied. 2. The cause of deterioration in the testis may be due to the clotting in the intercellular substances of the germinal epithelium as a result of the submaxillariectomy. 3. The atrophic change of germ cells in the testis of the submaxil lariectomized males may be due to the deterioration of intercellular substances in the germinal epithelium. 4. The secretion of submaxillary gland in the male guinea pigs seems to contain the anti-coagulate enzyme for the intercellular substances of germi nal epithelium. Literature cited Allanson, E. E., Hill, R. T. and McPhill, M. K J. Exp. Biol. 21: 348. Cutuly, E Proc. Soc. Exp. Biol. and Med. 47: Anat. Rec. 79. Supple 2: 16. -, Cutuly, E. C. and McCullagh, D. R Proc. Soc. Exp. Biol. and Med. 38: , McCullagh, D. R. and Cutuly E. C Am. J. Physiol. 119: 121. Nelson, W. O Anat. Rec. 67. Suppl 1: Proc. Soc. Exp. Biol. and Med. 38: 737. Suzuki, K Cytologia 22: Cytologia 24: Cytologia 26: 486. Walsh, E. L., Cuyler, W. K. and McCullagh, D. R Am. J. Physiol. 107: 548.