Histomorphometric Investigation of Changes In Adult Rat Testis After Experimental Vasectomy At Different Periods

Yıl 2020, Cilt: 25 Sayı: 2, 130 - 137, 13.06.2020

Alev Cumbul , Varol Şahintürk , Ünal Uslu

https://doi.org/10.21673/anadoluklin.666778

Öz

Aim: In previous studies, the causes of damage in seminiferous tubules were unclear; if germ cells and supporting cells were involved in the damage and what was the pattern of damage in terms of severity, extent and time course. The aim of this study is to qualitatively and quantitatively analyze the structural changes in seminiferous tubular epithelia thus explain the extent of structural damage of spermatogenic serial in rats.

Material and Methods: In this study, 8 weeks old, 48 male Wistar Albino rats were used. Vasectomy group underwent unilateral testes vasectomy, control group was subjected to abdominal incision. By using cardiac perfusion, left testes of all rats were removed after 1, 12 and 24 weeks (each group n=8). Specimens were immersed in Bouin's solution. Following routine histological procedures, 2 testes of each group stained with Masson trichrome and orcein for qualitative evaluation, 6 testes sectioned randomly (1/8 ratio) and stained with PAS+Hematoxylin. Morphometric data were obtained using stereological methods.

Results: Mean values of numerical density of spermatogonium, Sertoli and myoid cells in 1-12 week vasectomy groups were significantly higher than control. In contrast, amount of spermatids in vasectomy group was observed significantly lower. In vasectomy 24 week group, all cells in seminiferous tubules decreased significantly. In extracellular matrix, an increase was observed in the amount of collagen fibers, and was positively correlated with duration. In vasectomy 1 week group, giant cell formation was observed in lumen.

Conclusion: Two mechanisms may take part. Firstly, compensation mechanism can play role, due to paracrine and intercellular interactions in basal compartment. Secondly, seminal granulom of epididymis may increase the pressure of tubules, thus disrupts the spermatid-sertoli relationship. This may cause sloughing of spermatids in adluminal compartments.

Anahtar Kelimeler

Histomorphometry, vasectomy, testis, rat

Deneysel vazektominin farklı süreler sonrasında erişkin sıçan testisinde oluşturduğu değişikliklerin histomorfometrik olarak incelenmesi.

Öz

Amaç: Yapılan hayvan deneylerinde vazektomi sonrası spermatogenetik seride gözlenen ciddi hasarın oluşum mekanizması tam olarak aydınlatılamamıştır. Bu çalışmanın amacı erişkin sıçanların seminifer tübüldeki spermatogenetik serinin yapısal hasarını nitel ve nicel değişkenler yardımıyla mikroskopik olarak açıklamaya çalışmaktır.

Gereç ve Yöntem: Çalışmada 8 haftalık 48 adet erkek Wistar Albino sıçan kullanıldı. Deney gruplarına tek taraflı vazektomi uygulandı, kontrol grubundaki hayvanlara abdominal kesi yapılıp kapatıldı (her grup için n=8). Deney ve kontrol gruplarındaki sıçanların sol testisleri vazektomiden 1, 12 ve 24 hafta sonrasında perfüzyonla alınarak Bouin’s fiksatifine konuldu. Rutin histolojik takip işleminden sonra her gruptan ikişer adet testis trikrom masson ve orseinle boyanarak ışık mikroskopunda nicel olarak değerlendirildi. Altışar adet testis histomorfometrik inceleme için 1/8 oranında seri kesilerek PAS+Hematoksilenle boyandı. Stereolojik inceleme için Cavalieri ve optik parçalama+optik disektör sondaları kullanıldı.

Bulgular: 1 ve 12 haftalık vazektomi gruplarının spermatagonyum, Sertoli ve myoid hücrelerinin mm3 deki sayılarının kontrole göre arttığı (p<0,001), spermatid sayılarının ise azaldığı (p<0,001) saptandı. 24 haftalık vazektomi grubunda ise seminifer tübülü oluşturan hücrelerin azaldığı görüldü (p<0,001). Nicel değerlendirmede vazektomi sonrası süreye paralel olarak ara maddedeki kollajen lif miktarında artış ve 1 haftalık deney grubunda seminifer tübül lümeninde dev hücre oluşumu gözlemlenmiştir.

Sonuç: Bu bulgulara göre vazektomi sonrasında 12. haftaya kadar seminifer tübül adlüminal ve bazal kompartımanlarında iki farklı mekanizmanın çalıştığı görülmektedir. Bazal kompartımanda gerek parakrin gerekse hücreler arası bağlantılara bağlı bir telafi mekanizması çalışmaktadır. Epididimiste gelişen seminal granülomunun, seminifer tübül iç basıncını artırarak adlüminal kompartımanda spermatid sertoli ilişkisini bozduğu, bunun spermatidlerin olgunlaşamadan hızla dejenere olmasına neden olduğu düşünülmektedir.

Anahtar Kelimeler

Vazektomi, Testis, Sıçan, Histomorfometri

Kaynakça

• Amory JK, Male contraception. Fertility and sterility, 2016;106(6):1303-9.
• Tulsiani DRP and Abou-Haila A, Importance of male fertility control in family planning. Endocrine, metabolic & immune disorders drug targets, 2014;14(2):134-144.
• Weiske WH, Vasectomy. Andrologia, 2001;33(3): 125-134.
• Cook LA, Van Vliet HAAM, Lopez LM, Pun A, and Gallo MF, Vasectomy occlusion techniques for male sterilization. The Cochrane database of systematic reviews, 2014;3:CD003991.
• Makhlouf AA and Niederberger CS, Ensuring vasectomy success: what is the standard? Journal of andrology, 2006;27(5):637-640.
• Jacobs EJ, Anderson RL, Stevens VL, Newton CC, Gansler T, and Gapstur SM, Vasectomy and Prostate Cancer Incidence and Mortality in a Large US Cohort. Journal of clinical oncology, 2016;34(32):3880-5.
• Clarkson TB and N. J. Alexander NJ, Long-term vasectomy: effects on the occurrence and extent of atherosclerosis in rhesus monkeys. The Journal of clinical investigation, 1980;65(1):15-25.
• Bridges FS and Westerfield RC, Effects of vasectomy and exercise upon aortic extensibility, cholesterol deposition, and stainable tissue lipids. Biology of reproduction, .1984;31(2):296-302.
• Pasqualotto FF, Lucon AM, Sobreiro BP, Pasqualotto EB, and Arap S, The best infertility treatment for vasectomized men: assisted reproduction or vasectomy reversal? Revista do Hospital das Clinicas, 2004;59(5):312-5.
• Moss MB and Rosene DL, A perfusion-fixation procedure for the concurrent demonstration of Timm’s, horseradish peroxidase (HRP), and acethycholinesterase (AChE) histochemistry. The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 1984;32(10):1113-6.
• McFadden WC et al., Perfusion fixation in brain banking: a systematic review. Acta neuropathologica communications, 2019;7(1)146.
• Beach TG, Tago H, Nagai HT, Kimura H, McGeer PL, and McGeer EG, Perfusion-fixation of the human brain for immunohistochemistry: comparison with immersion-fixation. Journal of neuroscience methods, 1987;19(3):183-192.
• Gundersen HJ et al., The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. APMIS : acta pathologica, microbiologica, et immunologica Scandinavica, 1988;96(10):857-881. vol. 96, no. 10, pp. 857–881, Oct. 1988.
• Kohen. MC, Tatlipinar S, Cumbul A, and Uslu Ü, The effects of bevacizumab treatment in a rat model of retinal ischemia and perfusion injury. Molecular vision, 2018;24:239-250.
• Gundersen HJ and Jensen EB, “The efficiency of systematic sampling in stereology and its prediction. Journal of microscopy, 1987;147(3):229-263.
• Güleç A, Bakkalbaşı BÇ, Cumbul A, Uslu Ü, Alev B, and Yarat A, Effects of local platelet-rich plasma injection on the rate of orthodontic tooth movement in a rat model: A histomorphometric study,” American Journal of Orthodontics and Dentofacial Orthopedics, 2017;151(1)92-104.
• Yalcin-Ulker GM, Cumbul A, Duygu-Capar AG, Uslu Ü, and Sencift K, Preventive Effect of Phosphodiesterase Inhibitor Pentoxifylline Against Medication-Related Osteonecrosis of the Jaw: An Animal Study. Journal of Oral and Maxillofacial Surgery, 2017;75(11):2354-68.
• Yang ZW, Y. Guo Y, Lin L, X.-H. Wang. XH, J.-S. Tong JS, and G.-Y. Zhang GY, Quantitative (stereological) study of incomplete spermatogenic suppression induced by testosterone undecanoate injection in rats. Asian journal of andrology, 2004;6(4):291-297.
• Dobson CC, Reid O, Bennett NK, and McDonald SW, Effect of vasectomy on the seminiferous tubule boundary zone in the Albino Swiss rat. Clinical anatomy, 2000;13(4):277-286.
• Aydos K et al., Testicular effects of vasectomy in rats: an ultrastructural and immunohistochemical study. Urology, 1998;51(6):1051-56.
• Aitken H, Kumarakuru S, Reid O, Milne EW, Bennett NK, and McDonald SW, Degenerated tubules in the guinea pig testis after long-term vasectomy or sham operation. Clinical anatomy, 2000;13(1):6-10.
• Peng B, Zhang RD, Dai XS, Deng XZ, Wan Y, and Yang ZW, Quantitative (stereological) study of the effects of vasectomy on spermatogenesis in rhesus monkeys (Macaca mulatta). Reproduction, 2002;124(6):847-856.
• West DA, Chehval MJ, Winkelmann T, and Martin SA, Effect of vasovasostomy on contralateral testicular damage associated with unilateral vasectomy in mature and immature Lewis rats. Fertility and sterility, 2000;73(2):238-241.
• Anton E, Arrested apoptosis without nuclear fragmentation produced by efferent duct ligation in round spermatids and multinucleated giant cells of rat testis. Reproduction, 2003;125(6):879-887.
• Kong LS, Huang AP, Deng XZ, and Yang ZW, Quantitative (stereological) study of the effects of vasectomy on spermatogenesis in rabbits. Journal of anatomy, 2004;205(2):147-156.
• McDonald SW, Lockhart A, Gormal D, and Bennett NK, “Changes in the testes following vasectomy in the rat. Clinical anatomy, 1996;9(5):296-301.
• Ma L, Guo Y,Yuan Y, Li YG, Deng XZ, and Yang ZW, “Morphometric study of the testis and reproductive tract (including sperm granuloma) after vasectomy in mature rats. Asian journal of andrology, 2016;18(1):66-73.
• Lui WY, Mruk D, Lee WM, and Cheng CY, Sertoli cell tight junction dynamics: their regulation during spermatogenesis. Biology of reproduction, 2003;68(4):1087-97.