Oct 31, 2018

Recombinant production of rabbit β-Nerve Growth Factor and its biological effect on rabbit sperm

BioRxiv : the Preprint Server for Biology
Ana Sanchez-RodriguezRosa M Garcia-Garcia

Abstract

The neurotrophin β-Nerve Growth Factor (β-NGF) is flourishing as a protein with important roles in the ovulation induction process in induced-ovulation species but data in rabbits are still inconclusive, probably due to the species-specificity effect of the neurotrophin to trigger the ovulation. Moreover, β-NGF seems to have a role in sperm function. To clarify these functionalities we aimed, in the present research: 1) to newly synthesize a functional recombinant β-NGF from rabbit (rrβ-NGF), 2) to reveal differences in the amino acid sequence of rabbit β-NGF compared to other sequences of induced and spontaneous ovulator species, and 3) to assess the effects of rrβ-NGF on sperm viability and motility. The nucleotide sequence of NGF from rabbit prostate was sequenced by Rapid Amplification of cDNA Ends (RACE) and annotated in GenBank ([KX528686][1]). Then, rrβ-NGF was produced in CHO cells and purified by affinity chromatography. Western blot and MALDI-TOF analyses confirmed the correct identity of the recombinant protein. rrβ-NGF functionality was validated in PC12 cells through a successful dose-response effect along 8 days. The comparison of the amino acid sequences of NGF between rabbit and other species suggested some rele...Continue Reading

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Mentioned in this Paper

Research
Cell Motility
Western Blotting
NTRK1 wt Allele
Nerve Growth Factor
Ovulation
Reproduction
PC12 Cells
Evaluation of Sperm Viability
Sperm Cell

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