Date of Award

8-2013

Document Type

Thesis

Degree Name

Master of Science (MS)

College/School

College of Science and Mathematics

Department/Program

Biology

Thesis Sponsor/Dissertation Chair/Project Chair

John J. Gaynor

Committee Member

Lee H. Lee

Committee Member

Paul A. X. Bologna

Abstract

Jellyfish envenomations have increased drastically in recent years and have been known to cause a wide variety of reactions to the victim. These reactions can be as mild as minor skin irritation to the extreme of death. Many research groups have studied the wide variety of toxins contained within these organisms trying to understand the underlying mechanism that causes these dramatic changes noted to both the physiology and viability of cells. With no clear method for cnidocyst (nematocyst) extraction or lysis, these toxins cannot be recovered entirely, which would hinder research on potential mechanisms of action of the proteins found within jellyfish venom. We have investigated several different methods to isolate and purify cnidocysts as well as lysis of this organelle. Cnidocyst lysis has proven to be very difficult due to the rigid exterior capsule. Cnidocyst were isolated from sea nettles (iChrysaora quinquecirrha) collected from Bamegat Bay, NJ in the summer of 2012 or earlier. Freshly collected samples of C. quinquecirrha were autolysed in either artificial sea water or distilled water for 48 hrs at 4°C, homogenized then filtered through Miracloth and subjected to several rounds of centrifugation at varying speeds to purify cnidocyst samples. Further purification was achieved through a Percoll discontinuous gradient. Intact cnidocysts were placed in different buffer solutions at varying pH and subjected to either brief ultrasonic disruption or bead mill homogenization. Samples were then separated into soluble supernatant and pellet fractions and subjected to protein determination, microscopic imaging of cnidocyst lysis and SDS-PAGE analysis to determine both the number and size of proteins present in the venom. Lysed cnidocysts extracts and pellets were subjected to various hemolytic assays and we have detected hemolysin activity in the venom from C. quinquecirrha. Controls have demonstrated that this activity is due to a protein with the majority of the activity seen in the pellet fractions. It is hoped that with better isolation and extraction protocols, purer venom samples will be extracted that could lead to further research and potential development of pharmacologically active peptides. This may also provide novel strategies for the treatment or prevention of jellyfish envenomations.

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