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Tissue Specific Esterase Isozyme Banding Pattern in Nile Tilapia (oreochromis Niloticus)

Reza Md. Shahjahan
Laboratory of Animal genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka

Afroja Karim
Laboratory of Animal genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka

Rowshan Ara Begum
Laboratory of Animal genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka

Mohammad Shamimul Alam
Laboratory of Animal genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka

Anwara Begum
Laboratory of Animal genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka

Abstract/ Executive Summary:

The electrophoretic banding pattern of esterase isozymes were examined in eighteen different tissues of Nile tilapia after staining with α and β naphthyl acetate as substrate. The tissues were anterior-, mid-, tailmuscle (ventral and tip region), buccal muscle, stomach, fore-, mid-, hind-gut, liver, gill, heart, kidney, eye (lens), eye (iris), fore-, mid- and hind- brain. Altogether five bands named as Est-1^1.4, Est-2^1.1, Est-3¹, Est-4^0.62 and Est-5^0.25 were observed different relative mobility. Est-1 and Est- 5 denote the fastest and slowest band. Est-3 was present in all the tissues. All five bands were expressed in liver and stomach. Some of the esterase bands showed tissue and substrate specificity, where Est-1 was in fore-, mid- and hind- brain, Est-2 in hind gut, Est-5 in stomach, gill and heart was stained only with α naphthyl acetate. None of the band was expressed with β naphthyl acetate only.

Keywords: esterase, isozyme, Oreochromis, tissue

Location:

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Tilapia

Objectives:

The paper deals in polymorphic pattern of esterase isozymes in different body tissue of the Nile tilapia O. niloticus

Methodology:

The adult fish samples were collected from “M/S AL-AMIN MATSHA KHAMAR”, Narayangang in 1st May of 2007. The fishes were caught and taken live to the Laboratory of Animal Genetics and Molecular Biology, Department of Zoology, University of Dhaka, in plastic bags which was filled with water. The length of the specimen was 12cm, width 3.5cm and weight 22gm. The tissues studied were anterior muscle, mid muscle, tail muscle (ventral region), tail muscle (tip region), buccal muscle, stomach, foregut, midgut, hindgut, liver, eye (lens), eye (iris), gill, heart, kidney, forebrain, mid brain and hind brain. Altogether eighteen different tissues were removed and put into eppendorf tube as stock samples and were kept in a deep freeze at -800c. Gel preparation: 7.5% gel was prepared using 7.5 ml of Acrylamide-bisacrylamide (30:0.8) (Bio Basic Inc); 6.0 ml of 5X of TBE buffer; 0.150 ml of 10% ammonium persulphate (AMPS) (Fluka Biochemica); 0.140 ml of TEMED and 16.025 ml of water. Sample preparation: 0.015gm of each tissue sample was taken in a seperate eppendorf tube and squashed through electronic micro pester (Sigma) in 40 μl of lx Tris-Borate (MERCK) – EDTA (Sigma) buffer. 40 μl of Bromophenol blue (MERCK) in 1x TBE solution was added to each sample. The samples were centrifuged at 12000 rpm for 12 minutes at 250C, and 15 μl of supernatant was used in every slot of the gel. 7.5% Polyacrylamide gel electrophoresis (PAGE) was carried out to analyze. The gel was run at 120v constant voltage for at least 1 hour and 20 min until the tracking dye (bromophenol blue) reaches bottom of the gel. Then the gel was recovered by dismantling the cassettes/glass-plates sinking in water very cautiously. Staining: Elevation of esterase banding patterns was done in the presence of the two substrates, α and β naphthyl acetate in different tissues following basically the technique described by Johnson & Denniston (1964) and Steiner & Joslyn (1964). The gel was then taken in a staining tray and 30 ml of substrate mixture was poured onto it. It was kept in this mixture for 15 minutes at 25°C. After 15 minutes, substrate mixture was out poured and 30 ml of fast blue RR solution was added to the gel, it was incubated at 37°C for 25 minutes. Photograph of the gel was taken by a digital zoom lens camera laying the gel on a white background (Samsung Kenox-SHD lens, 3.2 Mega pixels). Scoring of bands: Bands were scored from the stained gel as the highest and lowest mobility of the band were numbered (Webb, 1964). Measuring total protein concentration by Lowry test: The principle behind the Lowry method is to determine protein concentration. Folin Ciocalteau was used as reagent solution, Bovin Serum Albumin (BSA) was used as a standard protein and optical density was measured at 660 nm wavelength. Dunn (1992) suggests concentrations ranging from 0.10 - 2 mg of protein per ml.

Source of Information: Univ. j. zool. Rajshahi Univ. Vol. 27, 2008 pp. 01-05 ISSN 1023-6104

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present study two esterase bands were found in all the three parts of the brain (fore, mid and hind) where Est-1 which was faintly stained and Est-3 medium deep stained in α naphthyl acetate. Only one esterase band Est-3 was found in β naphthyl acetate in all of the three parts of the brain. Banding pattern of esterases of different tissues has a good potential used in the identification of species. Between the two species the intestine of P. sutchi and P. pangasius possesses 4 and 6 bands, respectively. Furthermore, two species of Anabas (A. testudineus and A. oligolepis) was identified using esterase bands of liver, kidney, skeletal muscle, heart and egg. The findings in the present study may be extended to use as genetic marker in various fields of physiology, taxonomy and toxicology in Nile tilapia (O. niloticus)

Knowledge Management: Journal