Regulation of early spermatogenesis in the giant prawn Macrobrachium rosenbergii by a GCL homolog

Author:

Molcho Jonathan1,Albagly Dana1,Levy Tom23,Manor Rivka1,Aflalo Eliahu D14,Alfaro-Montoya Jorge5,Sagi Amir16

Affiliation:

1. Department of Life Sciences, Ben-Gurion University of the Negev , Beer-Sheva , Israel

2. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine , Stanford, CA , USA

3. Hopkins Marine Station, Stanford University , Pacific Grove, CA , USA

4. Department of Life Sciences, Achva Academic College , Arugot , Israel

5. Escuela de Ciencias Biológicas, Universidad Nacional , Heredia , Costa Rica

6. The National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev , Beer-Sheva , Israel

Abstract

Abstract The germ cell-less gene is crucial for gonad development in various organisms. Early interventions in its expression suggested a regulatory role at the mitotic stages of spermatogenesis, and its early knockout resulted in complete sterility in Drosophila. Genomic and transcriptomic data available for the catadromous giant prawn Macrobrachium rosenbergii enabled the identification of a germ cell-less homolog for this species, which we termed MroGCL (mRNA accession number OQ533056). An open reading frame containing 494 amino acids and a typical evolutionarily conserved BTB/POZ domain suggests possible protein–protein interaction functions in keeping with the Drosophila germ cell-less protein. Genomic mapping of MroGCL showed a full length of 120 896 bases. Analysis of the temporal expression of MroGCL showed constant expression in early prawn embryonic and larval stages, but a significant increase 10 days after metamorphosis when crucial sexual differentiation processes occur in prawns. In adult animals, high expression was detected in the gonads compared to the somatic tissues. RNAi-based knock-down experiments showed that both the silenced and control groups reached advanced spermatogenic stages, but that there was a significant decrease in the yield of spermatozoa in about half of the silenced animals. This finding supports our hypothesis that MroGCL is crucial for mitosis during early stage spermatogenesis. In conclusion, this study contributes to the understanding of crustacean gonad development and provides a stepping stone in the development of environmentally valuable sterile crustacean populations.

Funder

Ministry of Science & Technology

Publisher

Oxford University Press (OUP)

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