Targeted JAM-C deletion in germ cells by Spo11-controlled Cre recombinase

Abstract

SummaryMeiosis is a crucial process for the production of functional gametes. However, the biological significance of many genes expressed during the meiotic phase remains poorly understood, mainly because of the lethal phenotypes of the knockout mice. Functional analysis of such genes using the conditional knockout approach is hindered by the lack of suitable Cre transgenic lines. We describe here the generation of transgenic mice expressing Cre recombinase under the control of the meiotic Spo11 gene. Using LacZ-R26 loxP and EYFP-R26 loxP reporter mice, we show the specific expression and activity of Cre during meiosis in males and females. Spo11 Cre mice were then crossed with floxed Nbs1 and JAM-C mice to produce conditional knockouts. A strong reduction of Nbs1 and JAM-C protein levels was found in the testis. Although Nbs1-deleted mice developed minor gonadal abnormalities, JAM-C-knockout mice showed a spermiogenetic arrest, as previously described for the null mice. These results provide strong evidence that Spo11Cre transgenic mice represent a powerful tool for deleting genes of interest specifically in meiotic and/or in postmeiotic germ cells.

Neutrophil-Epithelial Interactions

“…65e67 An interesting feature of this family of proteins are the tissue-specific functions that seem to be governed by protein distribution patterns and may account for the wide variety of observed functions attributed to various JAMs. A few of the reported functions include regulation of epithelial barrier, 68,69 endothelial and epithelial cell migration, 70,71 spermatogenesis, 72 angiogenesis, 73 and leukocyte adhesion. 74 In addition, JAMs have been linked to roles in the pathophysiology of several disease processes, such as atherosclerosis, 75,76 cancer metastasis, 77 and IBD.…”

Essential Role of Sox2 for the Establishment and Maintenance of the Germ Cell Line

“…Rosa26 CreERT mice were obtained from Jackson Laboratories (Ann Harbor, http://www.jax.org). Spo11 Cre mice have been previously described in [21]. Homozygous YFPRosa26 loxP/loxP and LacZ-Rosa26 loxP/loxP mice were kindly provided by GianGiacomo Consalez (University of San Raffaele, Milan, Italy).…”

“…Meiotic and postmeiotic germ cells were obtained from adult testes and purified by elutriation, as previously described [21].…”

“…To understand if Sox2 might play a role also in oocyte development or in the initial phases of embryogenesis, before the initiation of embryonic transcription, we set out to delete Sox2 specifically in meiotic germ cells. To this end, we crossed Sox2 loxP/loxP with Sox2 lacZ;/þ ;Spo11 Cre , a deleter which expresses Cre in pachytene spermatocytes of the postnatal testis and in lepto-zygotene oocytes of fetal ovaries [21] ( Fig. 5A).…”

Current Frontiers and Perspectives in Cell Biology

“…Claudins (Furuse et al, 1998), (Furuse et al, 2002), (Nitta et al, 2003), (Turksen and Troy, 2002), (Tatum et al, 2010), (Nakano et al, 2009), (Gow et al, 1999), (Gow et al, 2004), (Ben-Yosef et al, 2003), (Tamura et al, 2008), (Simon et al, 1999), (Miyamoto et al, 2005), (Hou et al, 2009), (Angelow et al, 2008) Occludin (Furuse et al, 1993), (Traweger et al, 2002), (Sakakibara et al, 1997), (Saitou et al, 2000), (Furuse et al, 1998), (Du et al, 2010), (Runkle et al, 2011) MarvelD3 (Raleigh et al, 2010), (Steed et al, 2009) Tricellulin (Ikenouchi et al, 2005), (Ikenouchi et al, 2008) LSR (Masuda et al, 2011) JAMs (Hirabayashi and Hata, 2006), (Weber et al, 2007), (Kostrewa et al, 2001), (Laukoetter et al, 2007), (Pellegrini et al, 2011), (Ishida et al, 2003) …”