Characterization and Localization of 2 in Both the Testis and Ovary of the Japanese Flounder ()

Overview

Journal Animals (Basel)

Date 2020 Aug 30

PMID 32858799

Authors

Yuting Xiang

Yahui Wu

Haoran Zhang

Jikui Wu

Junling Zhang

Affiliations

Soon will be listed here.

Abstract

Although its function in mammalian gonads has been gradually recognized, the expression and function of calretinin (CALB2)-a Ca-binding protein-in the testis and ovary of fish are still unclear. Here, we identified the cDNA sequences of 2 in (); analyzed its gene structure and phylogenetic and syntenic relationship by bioinformatics; and investigated its tissue distribution and localization in the gonads by real-time PCR, western blotting, and immunohistochemistry. The 2 gene has 11 exons and 10 introns, and the full-length cDNA is 1457 bp, including an open reading frame (ORF) of 816 bp encoding 271 amino acids. The CALB2 of has a higher homology with (99%) compared with other species. The conserved synteny of 2 neighboring gene loci was also detected in fish. Real-time PCR showed that the expression of 2 mRNA is abundant not only in the brain, but also in the gonads, and exhibits a higher expression in the testis than in the ovary. Western blotting indicated that the CALB2 protein has a higher expression in the testis compared with the ovary. Immunohistochemistry demonstrated that the CALB2 protein appears in Leydig cells and the ovarian germ epithelium. These results reveal that 2 plays an important role in the gonads of .

References

Parmentier M, PASSAGE E, Vassart G, Mattei M . The human calbindin D28k (CALB1) and calretinin (CALB2) genes are located at 8q21.3----q22.1 and 16q22----q23, respectively, suggesting a common duplication with the carbonic anhydrase isozyme loci. Cytogenet Cell Genet. 1991; 57(1):41-3. DOI: 10.1159/000133111. View

Levanti M, Montalbano G, Laura R, Ciriaco E, Cobo T, Garcia-Suarez O . Calretinin in the peripheral nervous system of the adult zebrafish. J Anat. 2008; 212(1):67-71. PMC: 2423389. DOI: 10.1111/j.1469-7580.2007.00836.x. View

Germana A, Paruta S, Germana G, Ochoa-Erena F, Montalbano G, Cobo J . Differential distribution of S100 protein and calretinin in mechanosensory and chemosensory cells of adult zebrafish (Danio rerio). Brain Res. 2007; 1162:48-55. DOI: 10.1016/j.brainres.2007.05.070. View

Rogers J . Calretinin: a gene for a novel calcium-binding protein expressed principally in neurons. J Cell Biol. 1987; 105(3):1343-53. PMC: 2114790. DOI: 10.1083/jcb.105.3.1343. View

Shao C, Bao B, Xie Z, Chen X, Li B, Jia X . The genome and transcriptome of Japanese flounder provide insights into flatfish asymmetry. Nat Genet. 2016; 49(1):119-124. DOI: 10.1038/ng.3732. View

Stevens J, Rogers J . Chick calretinin: purification, composition, and metal binding activity of native and recombinant forms. Protein Expr Purif. 1997; 9(2):171-81. DOI: 10.1006/prep.1996.0677. View

Bertschy S, Genton C, Gotzos V . Selective immunocytochemical localisation of calretinin in the human ovary. Histochem Cell Biol. 1998; 109(1):59-66. DOI: 10.1007/s004180050202. View

Radi Z, Miller D . Immunohistochemical expression of calretinin in canine testicular tumours and normal canine testicular tissue. Res Vet Sci. 2005; 79(2):125-9. DOI: 10.1016/j.rvsc.2004.11.016. View

Schwaller B . Calretinin: from a "simple" Ca(2+) buffer to a multifunctional protein implicated in many biological processes. Front Neuroanat. 2014; 8:3. PMC: 3913827. DOI: 10.3389/fnana.2014.00003. View

10.

Pohl V, Van Rampelbergh J, Mellaert S, Parmentier M, Pochet R . Calretinin in rat ovary: an in situ hybridization and immunohistochemical study. Biochim Biophys Acta. 1992; 1160(1):87-94. DOI: 10.1016/0167-4838(92)90041-b. View

11.

Jadhao A, Deshpande K . Sexually dimorphic distribution of calcium-binding protein, calretinin in the preoptic area of the freshwater catfish, Clarias batrachus (Linn.). Neurosci Lett. 2014; 579:86-91. DOI: 10.1016/j.neulet.2014.07.020. View

12.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

13.

Lander E, Linton L, Birren B, Nusbaum C, Zody M, Baldwin J . Initial sequencing and analysis of the human genome. Nature. 2001; 409(6822):860-921. DOI: 10.1038/35057062. View

14.

Davidoff M, Middendorff R, Kofuncu E, Muller D, Jezek D, Holstein A . Leydig cells of the human testis possess astrocyte and oligodendrocyte marker molecules. Acta Histochem. 2002; 104(1):39-49. DOI: 10.1078/0065-1281-00630. View

15.

Foresta C, Mioni R . The role of calcium ions in rat Leydig cell steroidogenesis induced by atrial natriuretic peptide. Acta Endocrinol (Copenh). 1993; 128(3):274-80. DOI: 10.1530/acta.0.1280274. View

16.

Hack N, Wride M, Charters K, Kater S, Parks T . Developmental changes in the subcellular localization of calretinin. J Neurosci. 2000; 20(7):RC67. PMC: 6772258. View

17.

Meikle A, Liu X, STRINGHAM J . Extracellular calcium and luteinizing hormone effects on 22-hydroxycholesterol used for testosterone production in mouse Leydig cells. J Androl. 1991; 12(2):148-51. View

18.

Castro A, Becerra M, Manso M, Anadon R . Calretinin immunoreactivity in the brain of the zebrafish, Danio rerio: distribution and comparison with some neuropeptides and neurotransmitter-synthesizing enzymes. I. Olfactory organ and forebrain. J Comp Neurol. 2005; 494(3):435-59. DOI: 10.1002/cne.20782. View

19.

Bhoyar R, Jadhao A, Sivasubbu S, Singh A, Sabharwal A, Palande N . Neuroanatomical demonstration of calbindin 2a- and calbindin 2b-like calcium binding proteins in the early embryonic development of zebrafish: mRNA study. Int J Dev Neurosci. 2017; 60:26-33. DOI: 10.1016/j.ijdevneu.2017.03.012. View

20.

Barinka F, Druga R . Calretinin expression in the mammalian neocortex: a review. Physiol Res. 2010; 59(5):665-677. DOI: 10.33549/physiolres.931930. View