Pufferfish (Tetradontidae)

Jihea Choi and Bobby Adam Ryane

Chiral

What are Pufferfish?

Figure 1. Inflated pufferfish [6]

O
Pufferfish refers to a group of vertebrates that belongs in the Class Actinopterygii (bony fish),
in the Order Tetradontiformes, majority of them found in the Family Tetradontidae. There are 27
genera and 196 species of pufferfish. Puffers are found in marine, freshwater and brackish water
environments, however majority of their species are found mainly in marine environments,
ranging from the tropical and subtropical waters of Atlantic, Indian and Pacific Oceans. [1]
Puffer fish are famous for their poisonous nature. Mostly through bioaccumulation puffer fish
accumulates tetrodotoxin in their organs especially in the liver and ovaries [2]. There has been
evidence of production of tetrodotoxin in puffers but majority of the toxin is accumulated
through consumption of other organisms that contains the toxin. Bioacculmulation and biologic
magnification is the culprit of the large amounts of tetrodotoxin found in puffer fish [2]. Puffers
feed on invertebrates such as shellfish and algae and the tetrodotoxin that was present in the
food source gets accumulated
In puffer fish.

Chiral

OH

N
H

HO
OH

Chiral

HO

Chiral

+ Tetrodotoxin is an important compound to study since its considered a very

NH2

Chiral

Chiral

Chiral

strong poison found in the puffer fish, one of the major sea foods in
Japan.[11] How long Tetrodotoxin has been known is hard to pinpoint. The
earliest documentation was found from China The Book of Herbs from the
Emperor Shun Nung, 2838-2698 BC, time period. The book contained 365
drugs which were then classified into 3 categories according to toxicity.[12]
The isolation, purification and structural identification were elucidated and
published in the early 1960s.[13][14][15][16]

N
H

Figure 6. Tetrodotoxin structure with labelled functional groups

Colour

Functional Table 1.
Group
Colour

Red

Amine

Blue

Hydroxy

Green

Ether

Pharmacology

scheme in
Figure 6.

Toshio Narahashi in 1964 found that tetrodotoxin to act as a

Sodium channel blocker, but not effecting the potassium-carrying
system.[11] It blocks the action potential of the nerves found in a
number of animals (see figure 2).[17] Sodium channels are integral
proteins that create channels allowing for sodium to flow in or
out of a cell. The action potential of a cell, found in nerve cells,
is a rapid change in polarity across a membrane. This action
Figure 7. Schematic of Tetrodotoxin (left) and
space-filling model (right). [19]
potential thus allows for muscles to contract.[18] If the action
potential was blocked, this would cause known side-effects such as paralysis of voluntary muscles,
diaphragm (thus stopping a person from breathing), heart-rate, and sensation. The effects can be
felt at low dosages, 160 000 times stronger than cocaine.[12]

Puffers are also famous for their ability to puff their body up making them appear larger to
other organisms that may be a threat to them. Not only that the inflated body of puffer fish is very difficult to eat due to its round
nature and the increased size. Some puffers even have spines which makes it even harder for the predator to consume them [3].

Tetrodotoxin Toxicity
Tetrodotoxin found in puffer fish is very toxic to human
body. It causes neurotoxicity in human body by binding
to voltage gated sodium channels and preventing the
sodium ions to flow through the channels [4]. Thus, action
Figure 2. Blockage of ion channels by tetrodotoxin
potential is blocked and the nerve impulses are blocked,
meaning that bodily functions that require electrical nerve signals stops working. A person
ingesting tetrodotoxin usually ends up dying due to respiratory paralysis [5]. If a carnivorous fish
tries to prey on puffer and ingests tetrodotoxin, the toxin will make puffer to taste badly or
may even be toxic enough to kill the fish [3].

History

OH

O
Chiral

Tetrodotoxin

[7]

Active Site
NH2
O

+
NH2

HO

HN
NH

Other Organisms that has tetrodotoxin

OH

O
O

OH
OH

OH

NH2
N
H

HO
OH

N
H

HO

H2N

Another examples of an organism that utilizes tetrodotoxin is blue-ringed octopus and rough-skinned newt.
Tetrodotoxin can also be found in tetrodotoxin producing bacteria such as Vibrio alginolyticus.

Figure 8. Comparative structures of neosaxitoxin

(left) to tetrodotoxin (right).

Tetrodotoxin Derivatives

a.

O
O

Changing tetrodotoxin structure could potentially

decrease fatality of the toxin, while still keeping its
ability to cause lose of sensation. Eight
compounds were given with their respective
minimum lethal dose.20

OH

Figure 4. Vibrio alginolyticus

[9]

Figure 5. Rough-skinned newt

OH

Hypothesis on natural selection and puffer fish

Your hypothesis must include the process in which you think the particular
trait was selected for. You can provide evidence to back your hypothesis if
you wish too. Make sure the hypothesis is detailed and based on sound
scientific knowledge and is plausible.
References
[1]
[2]
[3]
[4]
[5]
[6]
[7]
[8]
[9]
[10]

Tetrodotoxin

8.22

Anhydrotetrodotoxin

4140

11-monoformylanhydrotetrodotoxin

3000

. 6,11-Diacetylanhydrotetrodotoxin

> 50 000

Tetrodonic acid

>>300 000

Methoxytetrodotoxin

341

. Ethoxytetrodotoxin

692

Deoxytetrodotoxin

84.5

. Tetrodaminotoxin

841

+
NH2

N
H

H3C

OH
OH
OH

O
N
H

HO

f.
OH

HO

N
H

g.

OH

N
H

h.
N
H

HO

OH

N
H

OH

NH2
N
H

HO
OH

O
H3C

OH

+
NH2

N
H

NH2
N
H

OH

HO
O

OH

N
H

OH

O
O

OH

H3C

NH2
N
H

OH

O
O

OH

+
NH2

N
H

H3C

N
H

NH2

e.
O

OH

d.

c.

OH

[10]

Derivative

OH

N
H

Minimum lethal dose (ug/kg)

N
H

HO

b.

NH2

OH

Figure 3. Blue-ringed Octopus [8]

Compounds such as neosaxitoxin exhibit sodium

channel blocking. When comparing the structure
of tetrodotoxin to the structure of neosaxitoxin
certain similarities can be seen. One noticeable
similarity is the guanidinium portion of the 2
compounds (the amine functional groups labeled
in red in figure 8). A hypothesis can thus be
reached that the guanidinium area of the
compounds are the active site, the area
responsible for interacting with the sodium
channel.

Similar in structure

H2N

N
H

Figure 9. Tetrodotoxin derivatives a. Anhydrotetrodotoxin b. 11monoformylanhydrotetrodotoxin c. 6,11-Diacetylanhydrotetrodotoxin d. Tetrodonic acid

e. Methoxytetrodotoxin f. Ethoxytetrodotoxin g. deoxytetrodotoxin h. Tetrodaminotoxin
Table 2. lethal amount of
tetrodotoxin derivatives
required compared to
tetrodotoxin

[11] Narahashi, T., Moore, J.W., Scott, W.R. Tetrodotoxin Blockage of Sodium Conductance Increase. J Gen Physio 1964; 47: 965-974.
[12] Kao, C.Y. Tetrodotoxin, Saxitoxin and Their Significance in the Study of Excitation Phenomena. Pharma Reviews 1966; 18: 997-1049.
[13] Goto, T., Kishi, Y., Takahashi, S., Hirata, Y. Tetrodotoxin. XI. Tetrahedron 1965; 21: 2059-2088.
[14] Mosher, H.S., Fuhrman, F.A., Buchwald, H.D., Fischer, H.G. Tarichatoxin-tetrodotoxin, a potent neurotoxin. Science 1964; 144: 1199-1110.
[15] Tsuda, K., Tachkikawa, R., Sakai, K., Tamura, C., Amakasu, O., Kawamura, M., Ikuma, S. On the structure of tetrodotoxin. Chem Pharm Bull 1964; 12: 642-645.
[16] Woodward, R.B. Structure of tetrodotoxin. Pure Appl Chem 1964; 9: 49-74.
[17] Ishihara, F. Uber die physiologischen wirkungen des fugutoxins. Mittheil Med Fak Tokio Univ 1918; 20: 375-426.
[18]
[19] Fozzard, H.A., Lipkind, G.M. The tetrodotoxin binding site is within the outer vestibule of the sodium channel. Marine Drugs 2010; 8(2): 219-234.
[20] Tsuda, K., Ikuma, S., Kawamura, M., Tachikawa, R., Sakai, K., Tamura, C., Amakasu, O. Tetrodotoxin. VII. On the structures of tetrodotoxin and its derivatives. Chem Pharm Bull 1964; 12(11): 1357-1374.