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European Microscopy Congress 2020 Abstract Database

IDENTIFICATION AND EXPRESSION OF NIEMANN PICK TYPE C2 LIKE GENE IN MALE REPRODUCTIVE TRACTS OF MACROBRACHIUM ROSENBERGII

IDENTIFICATION AND EXPRESSION OF NIEMANN PICK TYPE C2 LIKE GENE IN MALE REPRODUCTIVE TRACTS OF MACROBRACHIUM ROSENBERGII

Session

LSA.1 - Label-free life science imaging

Authors

Dr PPIYAPORN SURINLERT (1), Dr Chompoonut Sakulsret (2), Assoc.Prof.Dr Charoonroj Chotwiwattanakul (3), Assoc.Prof.Dr Somluk Asuvapongpattan (2), Assoc.Prof.Dr Wattana Weerachatyanukul (2)

Affiliations

1. Chulabhorn International College of Medicine, Thammasat University
2. Department of Anatomy, Faculty of Science, Mahidol University
3. Faculty of Science, Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Mahidol University

Keywords

Niemann pick type 2C (NPC2), M. rosenbergii, sperm maturation, cholesterol transportation

Abstract text

Macrobrachium rosenbergii is one of an economically important prawn species in tropical countries. A nauplii production sometimes has low as a poor fertilization rate.  Sperm fertility is considered as a remarkable criterion for a high fertilization rate success.  Niemann Pick type C2 (NPC2) is particularly known as a lipid binding protein and its significant role in an alteration of lipid composition on sperm membrane [1]. In crustacean, this protein was firstly cloned in P. monodon and the lipid binding ability was determined [2].   It is hypothesized that this protein might be the important male factor for sperm fertility in M. rosenbergii. A full cDNA sequence of NPC2 was identified from an available transcriptomic database using the nucleotide BLAST tool. The 1620 bps was encoded for 151 amino acids represented a putative signal peptide at N-terminus suggested it might be a secretory polypeptide. A conserved lipid recognition domain was characterized by CDD databases and a putative 3D structure was also determined with a similarity of mammal NCP2. The expression of MrNPC2 in male reproductive tract was analyzed by RT-PCR and In situ hybridization using a specific DNA probe. Highly MrNPC2 expression was detected in interstitial cells of testis. In vas deferens, it detected in a neighboring epithelium lining cells. Moreover, MrNPC2 protein was done by immunolocalization and it was detected in the epithelial lining of testis but was not detected in testicular sperm (Tsp), while it was highly accumulated in the epithelial lining of vas deferens and vas deferens sperm (Vsp). A correlation of MrNPC2 expression and sperm lipid composition implicated that this protein might be involved in sperm lipid modification. This MrNPC2 will be recombinantly expressed in E.coli, its function on lipid removal and sperm maturation in prawn will be determined in our next study.

References

1. Surinlert P, Asuvapongpatana S, Srakaew N, Daungchinda D, Setou M, Weerachatyanukul W, Aquaculture 456, (2016), p62-69.

2. Chotwiwatthanakun C, Sangatit J, Santimanawong W, Surinlert P, Prommoon J, Weerachatyanukul W, Vanichviriyakit R,  Mol Reprod Dev 15, (2016), p213-227.