Skip to main content
Log in

A missense mutation in LRR8 of RXFP2 is associated with cryptorchidism

  • Published:
Mammalian Genome Aims and scope Submit manuscript

Abstract

Using genome-wide mutagenesis with N-ethyl-N-nitrosourea (ENU), a mouse mutant with cryptorchidism was identified. Genome mapping and exon sequencing identified a novel missense mutation (D294G) in Relaxin/insulin-like family peptide receptor 2 (Rxfp2). The mutation impaired testicular descent and resulted in decreased testis weight in Rxfp2 DG/DG mice compared to Rxfp2 +/DG and Rxfp2 +/+ mice. Testicular histology of the Rxfp2 DG/DG mice revealed spermatogenic defects ranging from germ cell loss to tubules with Sertoli-cell-only features. Genetic complementation analysis using a loss-of-function allele (Rxfp2 ) confirmed causality of the D294G mutation. Specifically, mice with one of each mutant allele (Rxfp2 DG/−) exhibited decreased testis weight and failure of the testes to descend compared to their Rxfp2 +/− littermates. Total and cell-surface expression of mouse RXFP2 protein and intracellular cAMP accumulation were measured. Total expression of the D294G protein was minimally reduced compared to wild-type, but cell-surface expression was markedly decreased. When analyzed for cAMP accumulation, the EC50 was similar for cells transfected with wild-type and mutant RXFP2 receptor. However, the maximum cAMP response that the mutant receptor reached was greatly reduced compared to the wild-type receptor. In silico modeling of leucine rich repeats (LRRs) 7–9 indicated that aspartic acid 294 is located within the β-pleated sheet of LRR8. We thus postulate that mutation of D294 results in protein misfolding and aberrant trafficking. The ENU-induced D294G mutation underscores the role of the INSL3/RXFP2-mediated pathway in testicular descent and expands the repertoire of mutations known to affect receptor trafficking and function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Agoulnik AI (2005) Mouse mutants of relaxin, insulin-like 3 peptide and their receptors. Curr Med Chem Immunol Endocr Metab Agents 5:411–419

    Article  CAS  Google Scholar 

  • Balvers M, Spiess AN, Domagalski R, Hunt N, Kilic E et al (1998) Relaxin-like factor expression as a marker of differentiation in the mouse testis and ovary. Endocrinology 139:2960–2970

    Article  CAS  PubMed  Google Scholar 

  • Bogatcheva NV, Truong A, Feng S, Engel W, Adham IM et al (2003) GREAT/LGR8 is the only receptor for insulin-like 3 peptide. Mol Endocrinol 17:2639–2646

    Article  CAS  PubMed  Google Scholar 

  • Bogatcheva NV, Ferlin A, Feng S, Truong A, Gianesello L et al (2007) T222P mutation of the insulin-like 3 hormone receptor LGR8 is associated with testicular maldescent and hinders receptor expression on the cell surface membrane. Am J Physiol Endocrinol Metab 292:E138–144

    Article  CAS  PubMed  Google Scholar 

  • De Felice M, Di Lauro R (2004) Thyroid development and its disorders: genetics and molecular mechanisms. Endocr Rev 25:722–746

    Article  PubMed  Google Scholar 

  • Elder JS (1988) The undescended testis. Hormonal and surgical management. Surg Clin North Am 68:983–1005

    CAS  PubMed  Google Scholar 

  • Ferlin A, Simonato M, Bartoloni L, Rizzo G, Bettella A et al (2003) The INSL3-LGR8/GREAT ligand-receptor pair in human cryptorchidism. J Clin Endocrinol Metab 88:4273–4279

    Article  CAS  PubMed  Google Scholar 

  • Ferlin A, Zuccarello D, Zuccarello B, Chirico MR, Zanon GF et al (2008) Genetic alterations associated with cryptorchidism. JAMA 300:2271–2276

    Article  CAS  PubMed  Google Scholar 

  • Ferlin A, Zuccarello D, Garolla A, Selice R, Vinanzi C et al (2009) Mutations in INSL3 and RXFP2 genes in cryptorchid boys. Ann N Y Acad Sci 1160:213–214

    Article  CAS  PubMed  Google Scholar 

  • Gorlov IP, Kamat A, Bogatcheva NV, Jones E, Lamb DJ et al (2002) Mutations of the GREAT gene cause cryptorchidism. Hum Mol Genet 11:2309–2318

    Article  CAS  PubMed  Google Scholar 

  • Hartley BJ, Scott DJ, Callander GE, Wilkinson TN, Ganella DE et al (2009) Resolving the unconventional mechanisms underlying RXFP1 and RXFP2 receptor function. Ann N Y Acad Sci 1160:67–73

    Article  CAS  PubMed  Google Scholar 

  • Hsu SY, Nakabayashi K, Nishi S, Kumagai J, Kudo M et al (2002) Activation of orphan receptors by the hormone relaxin. Science 295:671–674

    Article  CAS  PubMed  Google Scholar 

  • Hsu SY, Nakabayashi K, Nishi S, Kumagai J, Kudo M et al (2003) Relaxin signaling in reproductive tissues. Mol Cell Endocrinol 202:165–170

    CAS  PubMed  Google Scholar 

  • Hutson JM, Baker M, Terada M, Zhou B, Paxton G (1994) Hormonal control of testicular descent and the cause of cryptorchidism. Reprod Fertil Dev 6:151–156

    Article  CAS  PubMed  Google Scholar 

  • Jensen MS, Toft G, Thulstrup AM, Henriksen TB, Olsen J et al (2010) Cryptorchidism concordance in monozygotic and dizygotic twin brothers, full brothers, and half-brothers. Fertil Steril 93:124–129

    Article  PubMed  Google Scholar 

  • Kobe B, Deisenhofer J (1995) A structural basis of the interactions between leucine-rich repeats and protein ligands. Nature 374:183–186

    Article  CAS  PubMed  Google Scholar 

  • Moran JL, Bolton AD, Tran PV, Brown A, Dwyer ND et al (2006) Utilization of a whole genome SNP panel for efficient genetic mapping in the mouse. Genome Res 16:436–440

    Article  CAS  PubMed  Google Scholar 

  • Nef S, Parada LF (1999) Cryptorchidism in mice mutant for Insl3. Nat Genet 22:295–299

    Article  CAS  PubMed  Google Scholar 

  • Overbeek PA, Gorlov IP, Sutherland RW, Houston JB, Harrison WR et al (2001) A transgenic insertion causing cryptorchidism in mice. Genesis 30:26–35

    Article  CAS  PubMed  Google Scholar 

  • Perry R, Heinrichs C, Bourdoux P, Khoury K, Szots F et al (2002) Discordance of monozygotic twins for thyroid dysgenesis: implications for screening and for molecular pathophysiology. J Clin Endocrinol Metab 87:4072–4077

    Article  CAS  PubMed  Google Scholar 

  • Pieper U, Eswar N, Webb BM, Eramian D, Kelly L et al (2009) MODBASE, a database of annotated comparative protein structure models and associated resources. Nucleic Acids Res 37:D347–D354

    Article  CAS  PubMed  Google Scholar 

  • Roh J, Virtanen H, Kumagai J, Sudo S, Kaleva M et al (2003) Lack of LGR8 gene mutation in Finnish patients with a family history of cryptorchidism. Reprod Biomed Online 7:400–406

    Article  CAS  PubMed  Google Scholar 

  • Scott DJ, Layfield S, Yan Y, Sudo S, Hsueh AJ et al (2006) Characterization of novel splice variants of LGR7 and LGR8 reveals that receptor signaling is mediated by their unique low density lipoprotein class A modules. J Biol Chem 281:34942–34954

    Article  CAS  PubMed  Google Scholar 

  • Scott DJ, Wilkinson TN, Zhang S, Ferraro T, Wade JD et al (2007) Defining the LGR8 residues involved in binding insulin-like peptide 3. Mol Endocrinol 21:1699–1712

    Article  CAS  PubMed  Google Scholar 

  • Siepka SM, Takahashi JS (2005) Forward genetic screens to identify circadian rhythm mutants in mice. Methods Enzymol 393:219–229

    Article  CAS  PubMed  Google Scholar 

  • Tomiyama H, Hutson JM, Truong A, Agoulnik AI (2003) Transabdominal testicular descent is disrupted in mice with deletion of insulinlike factor 3 receptor. J Pediatr Surg 38:1793–1798

    Article  PubMed  Google Scholar 

  • Toppari J, Kaleva M (1999) Maldescendus testis. Horm Res 51:261–269

    Article  CAS  PubMed  Google Scholar 

  • Yan Y, Scott DJ, Wilkinson TN, Ji J, Tregear GW et al (2008) Identification of the N-linked glycosylation sites of the human relaxin receptor and effect of glycosylation on receptor function. Biochemistry 47:6953–6968

    Article  CAS  PubMed  Google Scholar 

  • Zimmermann S, Steding G, Emmen JM, Brinkmann AO, Nayernia K et al (1999) Targeted disruption of the Insl3 gene causes bilateral cryptorchidism. Mol Endocrinol 13:681–691

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank the laboratory of Dr. David Beier for the SNP analysis. This work was supported by the Northwestern University Genomics Core and a Cancer Center Support Grant (NCI CA060553). This work was supported by the National Institutes of Health grants U01 HD043425-01(JLJ) and R01HD03706 (AIA).

Conflicts of interest

The authors have no conflicts of interest or financial ties to disclose.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to J. Larry Jameson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Harris, R.M., Finlayson, C., Weiss, J. et al. A missense mutation in LRR8 of RXFP2 is associated with cryptorchidism. Mamm Genome 21, 442–449 (2010). https://doi.org/10.1007/s00335-010-9291-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00335-010-9291-5

Keywords

Navigation