PUKYONG

GUIDE OAK 국가리포지터리 부경대학교

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PUKYONG Repository 대학원 공업화학과

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Alternative Title: Isolation and Characterization of Visceral Excitatory Neuropeptides from Cloudy Dogfish (Scyliorhinus torazame) Brain

Abstract: To investigate visceral excitatory neuropeptides of Chondrichthyes, peptides were purified from the brain of cloudy dogfish (Scyliorhinus torazame) using fish hindgut as the physiological assay system. Neuropeptides were purified from acetic acid brain extract of dogfish using reversed phase high performance liquid chromatography (HPLC) with sep-pak C18 cartridges. One of these visceral excitatory neuropeptides was determined by MALDI-TOF mass spectrometry and its amino acid sequence was determined by ESI-Q-TOF mass spectrometry. ESI-Q-TOF mass spectrometry showed that the peptide (Df-2) contained amino acid sequence of LESLVYEQLWPamide (1563Da). Threshold concentration of Df for the excitatory effect on the visceral segment of dogfish was 0.01㎍/㎖-1㎍/㎖ and minimum of 20min time was required for the complete recovery of intestine from the desensitization state brought by the Df-2 concentration of 1㎍/㎖.

URI: https://repository.pknu.ac.kr:8443/handle/2021.oak/3532
http://pknu.dcollection.net/jsp/common/DcLoOrgPer.jsp?sItemId=000001953420

Table Of Contents: Introduction = 1
Experimental Procedures = 13
1.Materials = 13
2.Methods = 13
2-1.Collection of specimens = 13
2-2. Extraction of neuropeptides = 13
2-3. Purification using sep-pak C18 cartridges = 14
2-4. High performance liquid chromatography = 14
2-5 Physiological assay = 15
2-6 Structure analysis = 18
Determination of molecular weight = 18
Determination of amino acid sequence = 18
2-7. Determination of physiological saline for dog fish = 19
2-8. Pharmacological effects on Df peptides = 19
Dose dependent response on dogfish hindgut tissue = 19
Time dependent response on dogfish hindgut tissue = 20
Results and Discussion = 21
Summary = 38
References = 39

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