Fish Physiology and Biochemistry

, Volume 44, Issue 3, pp 769–788 | Cite as

Analysis of the function of KIF3A and KIF3B in the spermatogenesis in Boleophthalmus pectinirostris

Article

Abstract

Spermatogenesis represents one of the most complicated morphological transformation procedures. During this process, the assembly and maintenance of the flagella and intracellular transport of membrane-bound organelles required KIF3A and KIF3B. Our main goal was to test KIF3A and KIF3B location during spermatogenesis of Boleophthalmus pectinirostris. We cloned complete cDNA of KIF3A/3B from the testis of B. pectinirostris by PCR and rapid amplification of cDNA ends (RACE). The predicted secondary and tertiary structures of B. pectinirostris KIF3A/3B contained three domains: (a) the head region, (b) the stalk region, and (c) the tail region. Real-time quantitative PCR (qPCR) results revealed that KIF3A and KIF3B mRNA were presented in all the tissues examined, with the highest expression seen in the testis. In situ hybridization (ISH) showed that KIF3A and KIF3B were distributed in the periphery of the nuclear in the spermatocyte and the early spermatid. In the late spermatid and mature sperm, the KIF3A and KIF3B mRNA were gradually gathered to one side where the flagella formed. Immunofluorescence (IF) showed that KIF3A, tubulin, and mitochondria were co-localized in different stages during spermiogenesis in B. pectinirostris. The temporal and spatial expression dynamics of KIF3A/3B indicate that KIF3A and KIF3B might be involved in flagellar assembly and maintenance at the mRNA and protein levels. Moreover, these proteins may transport the mitochondria resulting in flagellum formation in B. pectinirostris.

Keywords

KIF3A/3B Boleophthalmus pectinirostris Spermiogenesis Flagellar development 

Abbreviations

UTR

Untranslated region

RACE

Rapid amplification of cDNA ends

qPCR

Real-time quantitative polymerase chain reaction

ORF

Open reading frame

ISH

In situ hybridization

IF

Immunofluorescence

IFT

Intraflagellar transport

KIFs

Kinesin superfamily proteins

KAP3

Kinesin-associated protein 3

PBS

Phosphate buffered saline

OCT

Optimum cutting temperature

DEPC

Diethylpyrocarbonate

SSC

Sodium chloride

DIG

Digoxigenin

PFA

Paraformaldehyde

BSA

Albumin from bovine serum

TEM

Transmission electron microscopy

SD

Standard deviation

Notes

Acknowledgments

We are indebted to all members of the Fish Reproduction Physiology Laboratory at Ningbo University and the Sperm Laboratory at Zhejiang University for their enlightening discussions.

Funding information

This project was partially supported by the National Natural Science Foundation of China (No. 31272642), Scientific and Technical Project of Ningbo (No. 2015C110005), Ningbo Natural Science Foundation (No. 2016A610081), Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, K.C. Wong Magna Fund in Ningbo University, and Scientific Research Foundation of Graduate School of Ningbo University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10695_2017_461_MOESM1_ESM.docx (1.6 mb)
ESM 1 (DOCX 1602 kb)

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© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Key Laboratory of Applied Marine Biotechnology by the Ministry of Education, School of Marine SciencesNingbo UniversityNingboPeople’s Republic of China

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