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Esterase Variability in Different Tissues of Naked Neck Fowl (gallus Gallus Domesticus) of Bangladesh Using Polyacrylamide Gel Electrophoresis

Q. S. Akter
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

K. A. Muid
Laboratory of Animal Genetics and Molecular Biology, Department of Zoology, University of Dhaka, Dhaka-1000, Bangladesh

K. M. A. Tareq
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. A. M. Y. Khandoker
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Naked neck fowl (Gallus gallus domesticus) comprises one of the indigenous chicken populations in Bangladesh. The present investigation was conducted to study the polymorphic pattern of the estrase isozyme in different tissues of the fowl by polyacrylamide gel electrophoresis (PAGE) system. In this experiment, α-napthyle and β-napthyle acetate were used as substrate. Esterase variability of different samples of fore brain, mid brain, hind brain, heart brain, liver, testis, oesophagus, gizzard, bile ducts, proventiculus, small intestine, large intestine pectoral muscle, pelvic muscle, lung, eye, kidney, pancreas, body muscle and blood cells were examined. Altogether four esterase bands were observed and the bands were designated as Esterase Est-1 2.0, Est-21.0 Est-30.75 and Est-4o.15. Among the four bands Est-1 was the fastest and placed near to anode (+) electrode. Esterase showed tissue specific variation. Est-2 was found in almost all tissues, whereas Est-1 was observed only in liver. Est-3 and Est-4 were expressed exclusively in the nervous system especially in brain. Such findings may provide basic information to analyze the esterase activity in different tissues.

Keywords: Esterase, Isozyme, Naked neck fowl, Tissue

Location: Bangladesh Agricultural University, Mymensingh-2202

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Chicken

Objectives:

To study the polymorphic pattern of the esterase isozyme in different tissues.

Methodology:

Sample collection: The whole carcass of one Naked neck fowl was collected from Gazipur district in Bangladesh. The collected carcass was dissected to collect the required tissues viz. forebrain, midbrain, hindbrain, heart, liver, testis, oesophagus, gizzard, crop, proventiculous, small intestine, large intestine, pectoral muscle, pelvic muscle, lung, eye, kidney, pancreas, body muscle and blood. Separate special sterile bags were used for carrying the samples to avoid further contamination. The samples were than transported to the laboratory in an insulated box with ice to maintain the temperature around 4-6⁰C. In the laboratory, each sample was taken into eppendorf tube and kept in deep freeze at -80⁰C until further processing. Preparation of the sample: About 0.016gm different tissues of the Naked Neck fowl were taken in to an eppendrof tubes and squashed in 1 × Tris-Borate EDTA buffer adding about 40 ml. After squashing 2 × Bromophenol blue solution was added to each sample as the amount of 10 ml. The samples were then centrifuged at 12600 rpm for 15 min at 25⁰C. Ten microlitre of supernatant was used in each slot of the gel. Preparation of Gel: Seven and half percent (7.5%) gel was prepared using 7.5 ml of Acrylamide-bisacrylamide (30:0.8; Bio Basic Inc); 6.0 ml of 5 × of TBE buffer; 0.150 ml of 10% ammonium persulphate (AMPS; Fulka Biochemica); 0.140 ml of TEMED and 16 ml of sterilized distilled water. Electrophoresis of gel:The gel was run at 120V constant voltage for at least I hr and 20 min. until the tracking dye (bromophenol blue) reached to the bottom of gel. The gel was recovered by dismantling the glass plates sinking in water very cautiously. Staining and scoring of bands: The gel was taken in a staining tray and 30 ml of substrate mixture poured on to it for 15 min. at room temp. After 15 min, the substrate mixture was out poured and 30 ml of fast blue RR solution added to the gel and incubated at 37⁰C for 25 min. Photograph of the gel was taken by a digital zoom lens camera laying the gel on a white back ground (Samsung Kenox-SHD lens, 3.2 mega pixels). Bands were scored on the stained gel from the highest to the lowest mobility and numbered accordingly (Webb, 1964). Relative mobilities were estimated towards the commonly found band Est-2 as the band for unity. Akter et al. 53. Statistical analysis: The data generated from this experiment were entered in Microsoft Excel worksheet, organized and processed for further analysis. Data were analyzed using the general linear models procedure with the help of Statistical Package for Social Sciences (SPSS) 10/11.5.

Source of Information: J. Bangladesh Agril. Univ. 8(1): 51–55, 2010 ISSN 1810-3030

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results suggested tissue wise variation in the banding pattern of esterase and may be used as a good genetic marker in terms of species identification.

Knowledge Management: Journal