Expression characterization and functional implication of the collagen-modifying Leprecan proteins in mouse gonadal tissue and mature sperm

Abstract

The Leprecan protein family which includes the prolyl 3-hydroxylase enzymes (P3H1, P3H2, and P3H3), the closely related cartilage-associated protein (CRTAP), and SC65 (Synaptonemal complex 65, aka P3H4, LEPREL4), is involved in the post-translational modification of fibrillar collagens. Mutations in CRTAP, P3H1 and P3H2 cause human genetic diseases. We recently showed that SC65 forms a stable complex in the endoplasmic reticulum with P3H3 and lysyl hydroxylase 1 and that loss of this complex leads to defective collagen lysyl hydroxylation and causes low bone mass and skin fragility. Interestingly, SC65 was initially described as a synaptonemal complex-associated protein, suggesting a potential additional role in germline cells. In the present study, we describe the expression of SC65, CRTAP and other Leprecan proteins in postnatal mouse reproductive organs. We detect SC65 expression in peritubular cells of testis up to 4 weeks of age but not in cells within seminiferous tubules, while its expression is maintained in ovarian follicles until adulthood. Similar to bone and skin, SC65 and P3H3 are also tightly co-expressed in testis and ovary. Moreover, we show that CRTAP, a protein normally involved in collagen prolyl 3-hydroxylation, is highly expressed in follicles and stroma of the ovary and in testes interstitial cells at 4 weeks of age, germline cells and mature sperm. Importantly, CrtapKO mice have a mild but significant increase in morphologically abnormal mature sperm (17% increase compared to WT). These data suggest a role for the Leprecans in the post-translational modification of collagens expressed in the stroma of the reproductive organs. While we could not confirm that SC65 is part of the synaptonemal complex, the expression of CRTAP in the seminiferous tubules and in mature sperm suggest a role in the testis germ cell lineage and sperm morphogenesis.

Keywords

Leprecans - CRTAP - SC65 - testis - ovary - sperm morphology

^#a Current Address: Department of Molecular Medicine, Universita' di Pavia, Pavia, Italy

^#b Current Address: Department of Veterinary Integrative Biosciences, College of Veterinary Medicine and Biomedical Sciences, Texas A⨮M University, College Station, TX, USA

Publication History

Received: 05 October 2017

Accepted: 30 January 2018

Article published online:
10 May 2021

• References

• 1 Morello R, Bertin TK, Chen Y. et al. CRTAP is required for prolyl 3- hydroxylation and mutations cause recessive osteogenesis imperfecta. Cell 2006; 127: 291-304
• 2 Cabral WA, Chang W, Barnes AM. et al. Prolyl 3-hydroxylase 1 deficiency causes a recessive metabolic bone disorder resembling lethal/severe osteogenesis imperfecta. Nat Genet 2007; 39: 359-365
• 3 van Dijk FS, Nesbitt IM, Zwikstra EH. et al. PPIB mutations cause severe osteogenesis imperfecta. Am J Hum Genet 2009; 85: 521-527
• 4 Mordechai S, Gradstein L, Pasanen A. et al. High myopia caused by a mutation in LEPREL1, encoding prolyl 3-hydroxylase 2. Am J Hum Genet 2011; 89: 438-445
• 5 Hudson DM, Joeng KS, Werther R. et al. Post-translationally abnormal collagens of prolyl 3-hydroxylase-2 null mice offer a pathobiological mechanism for the high myopia linked to human LEPREL1 mutations. J Biol Chem 2015; 290: 8613-8622
• 6 Chen Q, Pearlman RE, Moens PB. Isolation and characterization of a cDNA encoding a synaptonemal complex protein. Biochem Cell Biol 1992; 70: 1030-1038
• 7 Fraune J, Schramm S, Alsheimer M. et al. The mammalian synaptonemal complex: protein components, assembly and role in meiotic recombination. Exp Cell Res 2012; 318: 1340-1346
• 8 Fossa A, Siebert R, Aasheim HC. et al. Identification of nucleolar protein No55 as a tumour-associated autoantigen in patients with prostate cancer. Br J Cancer 2000; 83: 743-749
• 9 Gaboreanu AM, Hrstka R, Xu W. et al. Myelin protein zero/P0 phosphorylation and function require an adaptor protein linking it to RACK1 and PKC alpha. J cell Biol 2007; 177: 707-716
• 10 Romcy-Pereira RN, Erraji-Benchekroun L, Smyrniotopoulos P. et al. Sleep-dependent gene expression in the hippocampus and prefrontal cortex following long-term potentiation. Physiol Behav 2009; 98: 44-52
• 11 Crockett DK, Fillmore GC, Elenitoba-Johnson KS. et al. Analysis of phosphatase and tensin homolog tumor suppressor interacting proteins by in vitro and in silico proteomics. Proteomics 2005; 5: 1250-1262
• 12 Gruenwald K, Castagnola P, Besio R. et al. Sc65 is a novel endoplasmic reticulum protein that regulates bone mass homeostasis. J Bone Mineral Res 2014; 29: 666-675
• 13 Heard ME, Besio R, Weis M. et al. Sc65-Null Mice Provide Evidence for a Novel Endoplasmic Reticulum Complex Regulating Collagen Lysyl Hydroxylation. PLoS Genet 2016; 12: e1006002
• 14 Hudson DM, Weis M, Rai J. et al. P3h3-null and Sc65-null Mice Phenocopy the Collagen Lysine Under-hydroxylation and Cross-linking Abnormality of Ehlers-Danlos Syndrome Type VIA. J Biol Chem 2017; 292: 3877-3887
• 15 Baldridge D, Lennington J, Weis M. et al. Generalized connective tissue disease in Crtap-/- mouse. PloS one 2010; 5: e10560
• 16 Chang W, Barnes AM, Cabral WA. et al. Prolyl 3-hydroxylase 1 and CRTAP are mutually stabilizing in the endoplasmic reticulum collagen prolyl 3-hydroxylation complex. Hum Mol Genet 2010; 19: 223-234
• 17 Skinner MK, Tung PS, Fritz IB. Cooperativity between Sertoli cells and testicular peritubular cells in the production and deposition of extracellular matrix components. J Cell Biol 1985; 100: 1941-1947
• 18 Irving-Rodgers HF, Hummitzsch K, Murdiyarso LS. et al. Dynamics of extracellular matrix in ovarian follicles and corpora lutea of mice. Cell Tissue Res 2010; 339: 613-624
• 19 Yang F, De La Fuente R, Leu NA. et al. Mouse SYCP2 is required for synaptonemal complex assembly and chromosomal synapsis during male meiosis. J Cell Biol 2006; 173: 497-507
• 20 Yuan L, Liu JG, Zhao J. et al. The murine SCP3 gene is required for synaptonemal complex assembly, chromosome synapsis, and male fertility. Mol Cell 2000; 5: 73-83
• 21 de Vries FA, de Boer E, van den Bosch M. et al. Mouse Sycp1 functions in synaptonemal complex assembly, meiotic recombination, and XY body formation. Genes Dev 2005; 19: 1376-1389
• 22 Bolcun-Filas E, Hall E, Speed R. et al. Mutation of the mouse Syce1 gene disrupts synapsis and suggests a link between synaptonemal complex structural components and DNA repair. PLoS Genet 2009; 5: e1000393
• 23 Bolcun-Filas E, Costa Y, Speed R. et al. SYCE2 is required for synaptonemal complex assembly, double strand break repair, and homologous recombination. J Cell Biol 2007; 176: 741-747
• 24 Schramm S, Fraune J, Naumann R. et al. A novel mouse synaptonemal complex protein is essential for loading of central element proteins, recombination, and fertility. PLoS Genet 2011; 7: e1002088
• 25 Hamer G, Wang H, Bolcun-Filas E. et al. Progression of meiotic recombination requires structural maturation of the central element of the synaptonemal complex. J Cell Sci 2008; 121: 2445-2451
• 26 He Z, Feng L, Zhang X. et al. Expression of Col1a1, Col1a2 and procollagen I in germ cells of immature and adult mouse testis. Reproduction 2005; 130: 333-341
• 27 Wyrobeck AJaB, Robert W. The Induction of Sperm-Shape Abnormalities in Mice and Humans. Chem Mutagens 1978; 257-285
• 28 Kawai Y, Hata T, Suzuki O. et al. The relationship between sperm morphology and in vitro fertilization ability in mice. J Reproduction Dev 2006; 52: 561-568