Pattern of expression of immune-relevant genes in the gonad of a teleost, the gilthead seabream (Sparus aurata L.)

Immunocytochemical Tools Reveal a New Research Field Between the Boundaries of Immunology and Reproductive Biology in Teleosts

“…The origin of the new Sertoli cells that restart spermatogenesis during the following reproductive cycle has been widely discussed and is still a matter of controversy. Our data demonstrated that after releasing the spermatozoa to the lumen of the tubules, the Sertoli cells remain forming the epithelium of the tubules where they phagocytose the remaining spermatozoa after spawning (Chaves-Pozo et al, 2005a, 2008a. Similarly when a post-spawning stage was induced by dietary intake of 17 -ethynilestradiol, the Sertoli cells released from the open www.intechopen.com…”

“…Regarding macrophages, the anti-Mcsfr revealed that this cell type was scattered in the testis and did not showed any cycling modifications in numbers, not during the first two reproductive cycles neither after 17 -ethynilestradiol dietary intake (Cabas et al, 2011;Chaves-Pozo et al, 2008a). Moreover, Sertoli cells also presented this molecule in their membrane, as other tissue specific macrophages such as those located in the spleen, liver or gills .…”

“…Interestingly, blood circulating acidophilic granulocytes only contained Il1 upon infection, however testicular acidophilic granulocytes constitutively produced Il1 , 2004a. Moreover, in the testis some spermatogonia, spermatocytes and scattered interstitial cells also produced Il1 (Chaves-Pozo et al, 2008a). In mammals Il1 has been described to be involved in germ cells proliferation in the testis (Syed et al, 1995), role that might also developed in fish as only the spermatogonia present in spermatogenesis and spawning stages, where the proliferative rate of spermatogonia are high, were immunostained with anti-Il1 antibody while at postspawning and involution stages, where the proliferative rate of spermatogonia were low, scarce spermatogonia were immunostained (Chaves-Pozo et al, 2005a, 2008b).…”

“…To reach this aim we have used several antibodies: one monoclonal antibody (G7) that is specific to gilthead seabream acidophilic granulocytes (Sepulcre et al, 2002) and three polyclonal antibodies: two against interleukin (Il)1 , obtained from the serum of an immunized rabbit with gilthead seabream recombinant Il1 or with the C-terminus of the gilthead seabream Il1 (RRHRIFKFLPPKPEVEGGEC), and one against macrophage colony stimulating factor receptor (Mcsfr), obtained from the serum of an immunized rabbit with a synthetic peptide corresponding to an internal epitope of the extracellular domain of the gilthead seabream Mcsfr (SLRVVRKEGEDYLLPC). The two last antibodies were produced by Pacific Immunology and characterized by Chaves-Pozo et al (2008a) and Mulero et al (2008), respectively. We have also used three commercial antibodies: two for the assessment of cell proliferation, anti-5'-Bromodesoxyuridine (BrdU) (Caltag) and anti-proliferating cell nuclear antigen (PCNA) (Dako) and an anti-B lymphocytes (Aquatic Diagnosis) which immunostained fish cells (Cabas et al, 2011;Chaves-Pozo et al, 2005a;Sepulcre et al, 2011).…”

“…Those enriched cell suspensions together with total testicular and head-kidney cell suspensions have been used in flow cytometry assays to asses phagocytosis (Esteban et al, 1998), reactive oxygen intermediates (ROI) production (Banati et al, 1994) and metalloproteinases activity (Leber and Balkwill, 1997). Combining those techniques, it has been demonstrated that headkidney acidophilic granulocytes infiltrate the gonad in response to gonadal and hormonal stimulus and showed completely different functional capabilities than their head-kidney counterparts (Chaves-Pozo et al, 2005b, 2008a: impaired phagocytic and ROI production activities although they were the majority of cells which were able to produce ROIs in the testis upon phorbol myristate acetate (PMA) stimulation (Chaves-Pozo et al, 2005b). Moreover, testicular acidophilic granulocytes also showed a completely different array of metalloproteinases activity than their head-kidney counterparts (Chaves-Pozo et al, 2008b).…”