Animal Reproduction (AR)
Animal Reproduction (AR)

Distribution of extracellular matrix related proteins in normal and cryptorchid ziwuling black goat testes

Hua Wang; Ligang Yuan; Juanjuan Song; Qianmei Wang; Yong Zhang

Downloads: 0
Views: 49


The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.


goat, cryptorchid, extracellular matrix, histochemistry, immunofluorescence


Baert Y, Stukenborg J-B, Landreh M, De Kock J, Jornvall H, Soder O, Goossens E. Derivation and characterization of a cytocompatible scaffold from human testis. Hum Reprod. 2015;30(2):256-67. PMid:25505010.

Berkholtz CB, Lai BE, Woodruff TK, Shea LD. Distribution of extracellular matrix proteins type I collagen, type IV collagen, fibronectin, and laminin in mouse folliculogenesis. Histochem Cell Biol. 2006;126(5):583-92. PMid:16758163.

Binsila KB, Selvaraju S, Ghosh SK, Parthipan S, Archana SS, Arangasamy A, Prasad JK, Bhatta R, Ravindra JP. Isolation and enrichment of putative spermatogonial stem cells from ram (Ovis aries) testis. Anim Reprod Sci. 2018;196:9-18. PMid:29861343.

Ezeasor D. Light and electron microscopical observations on the Leydig cells of the scrotal and abdominaltestes of naturally unilateral cryptorchid West African dwarf goats. J Anat. 1985;141:27-40. PMid:2867081.

Fayomi AP, Orwig KE. Spermatogonial stem cells and spermatogenesis in mice, monkeys and men. Stem Cell Res. 2018;29:207-14. PMid:29730571.

França LR, Godinho CL. Testis morphometry, seminiferous epithelium cycle length, and daily sperm production in domestic cats (Felis catus). Biol Reprod. 2003;68(5):1554-61. PMid:12606460.

Gur FM, Timurkaan S, Taskin E, Guven C, Gur HE, Senturk M, Dastan S, Nurdinov N, Unalan A, Cankut S, Tatyuz I. Thymoquinone improves testicular damage and sperm quality in experimentally varicocele‐induced adolescent rats. Andrologia. 2021;53(5):e14033. PMid:33660882.

Hafizuddin H, Karja NWK, Praharani L, Setiadi MA. Adiponectin and testosterone levels and their correlations with fertility in Anglo-Nubian x Etawah Grade Crossbred Bucks. Trop Anim Sci J. 2020;43:110-6.

Hayashi K, Hayashi M, Jalkanen M, Firestone JH, Trelstad RL, Bernfield M. Immunocytochemistry of cell surface heparan sulfate proteoglycan in mouse tissues. A light and electron microscopic study. J Histochem Cytochem. 1987;35(10):1079-88. PMid:2957423.

Heeren AM, van Iperen L, Klootwijk DB, de Melo Bernardo A, Roost MS, Gomes Fernandes MM, Louwe LA, Hilders CG, Helmerhorst FM, van der Westerlaken LA, Chuva de Sousa Lopes SM. Development of the follicular basement membrane during human gametogenesis and early folliculogenesis. BMC Dev Biol. 2015;15(1):4. PMid:25605128.

Hoflack G, Van den Broeck W, Maes D, Van Damme K, Opsomer G, Duchateau L, de Kruif A, Rodriguez-Martinez H, Van Soom A. Testicular dysfunction is responsible for low sperm quality in Belgian Blue bulls. Theriogenology. 2008;69(3):323-32. PMid:18023470.

Huang Y-F, Chen L-P, Zhao Y-J, Zhang H, Na R-S, Zhao Z-Q, Zhang JH, Jiang CD, Ma YH, Sun YW, e GX. Complete mitochondrial genome of Chuanzhong black goat in southwest of China (Capra hircus). Mitochondrial DNA A DNA Mapp Seq Anal. 2016;27(5):3063-4. PMid:25740215.

Ivell R, Alhujaili W, Kohsaka T, Anand-Ivell R. Physiology and evolution of the INSL3/RXFP2 hormone/receptor system in higher vertebrates. Gen Comp Endocrinol. 2020;299:113583. PMid:32800774.

Ivell R, Anand-Ivell R. Biological role and clinical significance of insulin-like peptide 3. Curr Opin Endocrinol Diabetes Obes. 2011;18(3):210-6. PMid:21415739.

Ivell R, Heng K, Anand-Ivell R. Insulin-like factor 3 and the HPG axis in the male. Front Endocrinol (Lausanne). 2014;5:1-8. PMid:24478759.

Jia J, Han L, Wang X, Liu W. Risk and regionalization of drought for Winter Wheat in Gansu Province. Arid Zo Res. 2019;36(6):1478-86.

Jia Y, Zhou J, Chang Y, An F, Li XW, Xu XY, Sun XL, Xiong CY, Wang JL. Effect of optimized concentrations of basic fibroblast growth factor and epidermal growth factor on proliferation of fibroblasts and expression of collagen: related to pelvic floor tissue regeneration. Chin Med J (Engl). 2018;131(17):17. PMid:30127219.

Jiang D, Fu X, Sheng Z. The diversity of structure and function of heparin sulfate proteoglycans via modification of some relative enzymes. Chinese J Pathophysiol. 2015;021:1020-5.

Li C, Yuan L, Zhang Y. The distribution of extracellular matrix proteins in small-tail han sheep epididymis in Plateau. Chinese J Anim Vet Sci. 2016;47:1565-73.

Li L, Zhang L, Zhang Z, Keyhani NO, Xin Q, Miao Z, Zhu Z, Wang Z, Qiu J, Zheng N. Comparative transcriptome and histomorphology analysis of testis tissues from mulard and Pekin ducks. Arch Tierzucht. 2020;63(2):303-13. PMid:32964101.

Lu T, Zou X, Liu G, Deng M, Sun B, Guo Y, Liu D, Li Y. A preliminary study on the characteristics of micrornas in ovarian stroma and follicles of chuanzhong black goat during estrus. Genes (Basel). 2020;55(9):970. PMid:32825655.

Madekurozwa MC, Booyse D. Seasonal changes in the immunolocalization of cytoskeletal proteins and laminin in the testis of the black-backed jackal (Canis mesomelas). J Vet Med Ser C Anat Histol Embryol. 2017;46(1):85-93. PMid:27477545.

McFarlane JR, Laslett A, De Kretser DM, Risbridger GP. Evidence that heparin binding autocrine factors modulate testosterone production by the adult rat Leydig cell. Mol Cell Endocrinol. 1996;118(1-2):57-63. PMid:8735591.

Minagawa I, Fukuda M, Ishige H, Kohriki H, Shibata M, Park EY, Kawarasaki T, Kohsaka T. Relaxin-like factor (RLF)/insulin-like peptide 3 (INSL3) is secreted from testicular Leydig cells as a monomeric protein comprising three domains B–C–A with full biological activity in boars. Biochem J. 2012;441(1):265-73. PMid:21899516.

Moore BC, Hamlin HJ, Botteri NL, Lawler AN, Mathavan KK, Guillette LJ Jr. Posthatching development of Alligator mississippiensis ovary and testis. J Morphol. 2010;271(5):580-95. PMid:20013789.

Murdock MH, David S, Swinehart IT, Reing JE, Tran K, Gassei K, Orwig KE, Badylak SF. Human testis extracellular matrix enhances human spermatogonial stem cell survival in vitro. Tissue Eng Part A. 2019;25(7-8):663-76. PMid:30311859.

Park MH, Park JE, Kim MS, Lee KY, Hwang JY, Yun JI, Choi JH, Lee E, Lee ST. Effects of extracellular matrix protein-derived signaling on the maintenance of the undifferentiated state of spermatogonial stem cells from porcine neonatal testis. Asian-Australas J Anim Sci. 2016;29(10):1398-406. PMid:26954208.

Piprek RP, Kolasa M, Podkowa D, Kloc M, Kubiak JZ. Transcriptional profiling validates involvement of extracellular matrix and proteinases genes in mouse gonad development. Mech Dev. 2018;149:9-19. PMid:29129619.

Pitia AM, Uchiyama K, Sano H, Kinukawa M, Minato Y, Sasada H, Kohsaka T. Functional insulin-like factor 3 (INSL3) hormone-receptor system in the testes and spermatozoa of domestic ruminants and its potential as a predictor of sire fertility. Anim Sci J. 2017;88(4):678-90. PMid:27592693.

Qianmei W, Ligang Y, Chenye L, Hongzao Y, Shaoyu C. Distribution of sex hormone receptors in cryptorchidism and normal tests of ziwuling black goat. Acta Vet Zootech Sin. 2020;51:1455-565.

Qin W, Wang B, Yang L, Yuan Y, Xiong X, Li J, Yin S. Molecular cloning, characterization, and function analysis of the AMH gene in Yak (Bos grunniens) Sertoli cells. Theriogenology. 2021;163:1-9. PMid:33476894.

Santamaria L, Martinez-Onsurbe P, Paniagua R, Nistal M. Laminin, type IV collagen, and fibronectin in normal and cryptorchid human testes. An immunohistochemical study. Int J Androl. 1990;13(2):135-46. PMid:1971616.

Siu MKY, Cheng CY. Extracellular matrix and its role in spermatogenesis. Adv Exp Med Biol. 2008;636:74-91. PMid:19856163.

Soito ICS, Favorito LA, Costa WS, Sampaio FJB, Cardoso LEM. Extracellular matrix remodeling in the human gubernaculum during fetal testicular descent and in cryptorchidic children. World J Urol. 2011;29(4):535-40. PMid:21626117.

Taranta A, Teti A, Stefanini M, D’Agostino A. Immediate cell signal induced by laminin in rat Sertoli cells. Matrix Biol. 2000;19(1):11-8. PMid:10686421.

Tung PS, Fritz IB. Interactions of sertoli cells with myoid cells in vitro. Biol Reprod. 1980;23(1):207-17. PMid:7191336.

Verkauskas G, Malcius D, Dasevicius D, Hadziselimovic F. Histopathology of unilateral cryptorchidism. Pediatr Dev Pathol. 2019;22(1):53-8. PMid:30012073.

Walker WH. Molecular mechanisms of testosterone action in spermatogenesis. Steroids. 2009;74(7):602-7. PMid:19095000.

Wang R, Okamoto M, Xing X, Crawford NM. Microarray analysis of the nitrate response in arabidopsis roots and shoots reveals over 1,000 rapidly responding genes and new linkages to glucose, trehalose-6-phosphate, iron, and sulfate metabolism. Plant Physiol. 2003;132(2):556-67. PMid:12805587.

Xing J, Bai Z. Is testicular dysgenesis syndrome a genetic, endocrine, or environmental disease, or an unexplained reproductive disorder? Life Sci. 2018;194:120-9. PMid:29183799.

Yuan L, Zhu J, Gu L, Yan Z. The histologic characters of testis in aging yak. Histol Characters Testis Aging Yak. 2015;46:855‎–‎62.

Yuan L, Qu Y, Li C. The histologic and ultrastructural characteristics of the Bactrian camel testis in cryptorchidism. Chinese J Anim Vet Ences. 2016;47:993-1000.

Yuan L, Lu Y, Tao J, Zhang Y. Comparison of histochemical and ultrastructural characteristics of extracellular matrix components in the scrotal and cryptorchid testes of the Bactrian camel. Acta Theriologica Sinica. 2017a;37:189-99.

Yuan L, Zhang Y, Li C, Cheng X. Comparison of distribution characteristics of extracellular matrix components in the testis of the Tibetan sheep and the small-tail Han sheep from plateau. Acta Anat Sin. 2017b;48:179-86.

Yuan L, Wang H, Wang Q, Li C, Yang D. INSL-3 protein expression in normal and cryptorchid testes of Ziwuling black goats. Reprod Domest Anim. 2021;56(5):725-35. PMid:33544931.

Zhao H, Nie J, Zhu X, Lu Y, Liang X, Xu H, Yang X, Zhang Y, Lu K, Lu S. In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig. J Vet Sci. 2018;19(5):592-9. PMid:29929354.

Zhu J, Gu L, Yan Z, Tian D, Yuan L. Morphological changes of testicular seminiferous tubules and Leyig cells of yak in different ages. Gansu Nongye Daxue Xuebao. 2014;4:18-22.

Submitted date:

Accepted date:

62952224a95395656b44ffb4 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections