An insight into testis and gubernaculum dynamics of INSL3–RXFP2 signalling during testicular descent in the dog
S. Arrighi A C , G. Bosi A , D. Groppetti B , M. Aralla A and F. Cremonesi BA Department of Veterinary Sciences and Technologies for Food Safety, Laboratory of Anatomy, Università degli Studi di Milano, Italy.
B Department of Veterinary Clinical Science, Reproduction Unit, Faculty of Veterinary Medicine, Università degli Studi di Milano, Italy.
C Corresponding author. Email: silvana.arrighi@unimi.it
Reproduction, Fertility and Development 22(5) 751-760 https://doi.org/10.1071/RD09260
Submitted: 23 October 2009 Accepted: 24 November 2009 Published: 7 April 2010
Abstract
Insulin-like 3 (INSL3) plays a prominent role in male development and is supposed to induce the growth of the gubernaculum testis (g.t.), thus being directly involved in testicular descent in humans and rodents. This happens through activation of the RXFP2 receptor (GREAT or LGR8). The INSL3–RXFP2 complex is reputed to play an additional paracrine role in the testis, possibly acting as part of an autocrine feedback loop. The present work provides evidence of the immunolocalisation of INSL3 in the Leydig cells of canine fetuses and of the expression of RXFP2 receptor in different tissues of the g.t. of the same specimens. RXFP2 was localised at the cell membrane of g.t. muscle and connective cells, as well as in the epithelial cells of the developing excurrent ducts. Notably, RXFP2 immunoreactivity of the g.t. was limited to fetuses at ~35–45 days of gestation, which is also the fetal period when the endocrine compartment of the dog testis is active endocrinologically, as confirmed by the anti-P450c17 and anti-INSL3 immunoreactivities of the fetal Leydig cells, and by anti-Müllerian hormone immunoreactivity of the Sertoli cells. The same immunoreactivities were also evaluated in the testes of cryptorchid dogs of different ages. RXFP2 immunoreactivity was absent from genital tracts of cryptorchid testes and g.t. remnants.
Additional keywords: anti-Müllerian hormone, cryptorchidism, GREAT, RLF, testicular ligaments.
Acknowledgements
Prof. Alan J. Conley, U. C. Davis School of Veterinary Medicine, USA is warmly acknowledged for kindly providing the anti-P450c17 antibodies and for his skilful suggestions during the preparation of the manuscript. Mr. Paolo Stortini (VSA, University of Milan) is gratefully acknowledged for his experienced and competent technical support. This work was supported by the University of Milan (PUR 2008). Part of this work was published in a special issue of Veterinary Research Communications (33, Suppl 1:S67–S71, 2009), containing a selection of abstracts from the LXII Meeting of the Italian Society of Veterinary Sciences (S.I.S.Vet.), held in San Benedetto del Tronto (Italy) in September 2008.
Adham, I. M. , Burkhardt, E. , Benahmed, M. , and Engel, W. (1993). Cloning of a cDNA for a novel insulin-like peptide of the testicular Leydig cells. J. Biol. Chem. 268, 26 668–26 672.
| PubMed | CAS |
Kawamura, K. , Kumagai, J. , Sudo, S. , Chun, S. Y. , and Pisarska, M. , et al. (2004). Paracrine regulation of mammalian oocyte maturation and male germ cell survival. Proc. Natl. Acad. Sci. USA 101, 7323–7328.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Klonisch, T. , Kauffold, J. , Steger, K. , Bergmann, M. , Leiser, R. , Fischer, B. , and Hombach-Klonisch, S. (2001). Canine relaxin-like factor: unique molecular structure and differential expression within reproductive tissues of the dog. Biol. Reprod. 64, 442–450.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Kothapalli, K. , Kirkness, E. , Pujar, S. , Van Wormer, R. , and Meyers-Wallen, V. N. (2005). Exclusion of candidate genes for canine SRY-negative XX sex reversal. J. Hered. 96, 759–763.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Kubota, Y. , Temelcos, C. , Bathgate, R. A. , Smith, K. J. , Scott, D. , Zhao, C. , and Hutson, J. M. (2002). The role of insulin 3, testosterone, Müllerian inhibiting substance and relaxin in rat gubernacular growth. Mol. Hum. Reprod. 8, 900–905.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Kumagai, J. , Hsu, S. Y. , Matsumi, H. , Roh, J. S. , Fu, P. , Wade, J. D. , Bathgate, R. A. , and Hsueh, A. J. (2002). INSL3/Leydig insulin-like peptide activates the LGR8 receptor important in testis descent. J. Biol. Chem. 277, 31 283–31 286.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Kutzler, M. A. , Yeager, A. E. , Mohammed, H. O. , and Meyers-Wallen, V. N. (2003). Accuracy of canine parturition date prediction using fetal measurement obtained by ultrasonography. Theriogenology 60, 1309–1317.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Nef, S. , and Parada, L. F. (1999). Cryptorchidism in mice mutant for insl3. Nat. Genet. 22, 295–299.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Overbeek, P. A. , Gorlov, I. P. , Sutherland, R. W. , Houston, J. B. , Harrison, W. R. , Boettger-Tong, H. L. , Bishop, C. E. , and Agoulnik, A. I. (2001). A transgenic insertion causing cryptorchidism in mice. Genesis 30, 26–35.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Pusch, W. , Balvers, M. , and Ivell, R. (1996). Molecular cloning and expression of the relaxin-like factor from the mouse testis. Endocrinology 137, 3009–3013.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Scott, D. J. , Fu, P. , Shen, P. J. , Gundlach, A. , Layfield, S. , Riesewijk, A. , Tomiyama, H. , Hutson, J. M. , Tregear, G. W. , and Bathgate, R. A. (2005). Characterization of the rat INSL3 receptor. Ann. N. Y. Acad. Sci. 1041, 13–16.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Shenker, N. S. , Huynh, J. , Farmer, P. J. , and Hutson, J. M. (2006). A new role for androgen in testicular descent: permitting gubernacular cell proliferation in response to the neuropeptide, calcitonin gene-related peptide. J. Pediatr. Surg. 41, 407–412.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Sironi, L. , Ramelli, P. , Lazzari, B. , Groppetti, D. , Lange Consiglio, A. , Mariani, P. , and Cremonesi, F. (2007). Analysis of rxfp2 (lgr8) as candidate gene for canine cryptorchidism. Reprod. Domest. Anim. 42(Suppl. 2), 82.
Sugita, Y. , Paxton, G. A. , Hasthorpe, S. , and Hutson, J. M. (1997). Does calcitonin gene-related peptide act as a chemoattractant for rat gubernacular cells? J. Pediatr. Surg. 32, 15–17.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Truong, A. , Bogatcheva, N. V. , Schelling, C. , Dolf, G. , and Agoulnik, A. I. (2003). Isolation and expression analysis of the canine insulin-like factor 3 gene. Biol. Reprod. 69, 1658–1664.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Virtanen, H. E. , and Toppari, J. (2008). Epidemiology and pathogenesis of cryptorchidism. Hum. Reprod. Update 14, 49–58.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Yamanaka, J. , Metcalfe, S. A. , Hutson, J. M. , and Mendelsohn, F. A. (1993). Testicular descent. II. Ontogeny and response to denervation of calcitonin gene-related peptide receptors in neonatal rat gubernaculum. Endocrinology 132, 280–284.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Yates, D. , Hayes, G. , Heffernan, M. , and Beynon, R. (2003). Incidence of cryptorchidism in dogs and cats. Vet. Rec. 152, 502–504.
| PubMed | CAS |
Zimmermann, S. , Steding, G. , Emmen, J. M. , Brinkmann, A. O. , Nayernia, K. , Holstein, A. F. , Engel, W. , and Adham, I. M. (1999). Targeted disruption of the Insl3 gene causes bilateral cryptorchidism. Mol. Endocrinol. 13, 681–691.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
