Aquaculture Europe 2023

September 18 - 21, 2023


Add To Calendar 21/09/2023 16:15:0021/09/2023 16:30:00Europe/ViennaAquaculture Europe 2023SPERMATOGONIAL PROLIFERATION AND APOPTOSIS IN PREPUBERTAL MEAGRE Argryrosomus regius TREATED WITH RECOMBINANT FOLLICLE STIMULATING HORMONE, AND COMPARISON WITH ADULTSStolz 1The European Aquaculture Societywebmaster@aquaeas.orgfalseDD/MM/YYYYaaVZHLXMfzTRLzDrHmAi181982


R. Zupa1*, N. Duncan2 , I. Giménez3 , C. C. Mylonas4, C. Pousis1 , G. Ventriglia1 ,  L. Valentini5 , C. Varvara1 , A. Corriero1


1 Department of Veterinary Medicine, University of Bari Aldo Moro, Valenzano 70010 ,  Bari (Italy)

2Institute of Agrifood Research and Technology, 43540 La Ràpita, Tarragona, Spain

3Rara Avis Biotec, S. L., Calle Moratín 17, 46002 Valencia, Spain

4Institute of Marine Biology, Biotechnology and Aquaculture, Hellenic Center for Marine Research, Heraklion 71003, Crete , Greece

5DiMePRe-J, University of Bari Aldo Moro, Valenzano 70010, Bari (Italy)




 The meagre Argyrosomun regius (Asso, 1801) is a promising aquaculture species and advancing puberty using recombinant gonadotropins could shorten the generation time for selective breeding programs (Zupa et al., 2023) . The aim of this study was to assess the effects of recombinant follicle stimulating hormone (rFsh) administration on spermatogonial proliferation and apoptosis in prepubertal meagre reared in indoor tanks  through a comparison with adult fish reared in sea cages.

Material and Methods

 Prepubertal meagre males (18-months old) reared in indoor tanks at IRTA (La Ràpita, Spain) underwent a six-weeks treatment with increasing doses of rFsh (week 0: 6 µg/kg; week 1: 9 µg/kg; week 2 to week 6: 12 µg/kg); c ontrol  prepubertal males were injected weekly with 1 mL of saline solution. Prepubertal fish samplings took place before the treatment (week 0;  control fish, N = 6) and after 6 weeks of treatment (week 6; control fish, N = 9 and rFsh-treated fish, N = 4 ). A dult males (6-years old) belonging to a commercial stock reared in sea cages in the Gulf of Taranto (Ionian Sea, Italy) were sacrificed during  early (March-April 2021; N = 7) and advanced (June 2021; N = 4) phases of spermatogenesis . T estis samples were fixed in Bouin’s solution and embedded in paraffin wax. Deparaffinized sections were stained with hematoxylin-eosin ; proliferating spermatogonia were  identified through  the immunohistochemical detection of the proliferating cell nuclear antigen (PCNA) ;  apoptotic germ cells were identified through  the TUNEL method .

Results and Discussion

 The rFsh-treated fish had larger testes compared to both control groups, had  larger seminiferous tubules that contained all stages of spermatogenesis and  had more abundant luminal spermatozoa (Fig. 1a, b). The testes of adult fish sampled in March-April were in active spermatogenesis with all germ cell types in the germinal epithelium and luminal spermatozoa (Fig. 1c );  while  in June, all adults were  fully mature, showing residual spermatogenetic activity in a thin germinal epithelium and plenty of luminal spermatozoa (Fig. 1d ).  Anti-PCNA immunostaining was observed in the nuclei of single spermatogonia, spermatogonia in cysts and primary spermatocytes (Fig. 1e ), but only  single  spermatogonia were considered for quantitative analysis. The TUNEL reaction labelled spermatogonia and spermatocytes (Fig. 1f). Fish treat ed  with rFsh  showed a significant decrease of proliferating  and apoptotic  single spermatogonia. In adults, spermatogonial proliferation was significantly higher during t he early phase of spermatogenesis  compared  with the advanced phase in June,  and  apoptosis significantly increased  from the early to the  advanced phase of spermatogenesis (Table 1).

The treatment with rFsh stimulated spermatogenesis advancement in prepubertal meagre and induced a significant reduction in spermatogonial proliferation and apoptosis. In adult fish, germ cell apoptosis was low during  the  early spermatogenesis phase and increased during the advanced spermato genesis phase. This observation confirms that apoptosis plays a major role in regulating germ cells/Sertoli cells ratio and in preventing aberrant germ cell development during spermatogenesis in adult fish (Prisco et al., 2003; Zupa et al., 2013). Moreover, t he present data  support our previous hypothesis that  in prepubertal  meagre apoptosis is involved in the inhibition of spermatogonial survival and progress towards meiosis (Zupa et al., 2023).

Financial grants provided by the EU Programme H2020 (GA 862658, NewTechAqua), and by the Apulian Region to R.Z. (POR PUGLIA FESR-FSE 2014/2020 – Asse X-Azione 10.4 – REFIN) and to C.V. (POC PUGLIA FESR-FSE 2014/2020-RIPARTI).


 Prisco, M. et al. (2003). Apoptosis during spermatogenesis in the spotted ray Torpedo marmorata . Mol. Reprod. Dev. 64:341–348.

 Zupa, R. et al. (2013). Comparative analysis of male germ cell proliferation and apoptosis in wild and captive Atlantic bluefin tuna (Thunnus thynnus L.). J. Appl. Ichthyol. 29:71–81.

Zupa, R. et al. (2023). Male germ cell proliferation and apoptosis in sexually immature meagre Argyrosomus regius (Asso, 1801) treated with recombinant follicle stimulating hormone. Sci. Rep. 13:7013.