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Cephalochordata (“lancelets”) - PowerPoint PPT Presentation


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Phylum Chordata. Cephalochordata (“lancelets”). Urochordata (“tunicates). Agnatha (jawless fish, e.g. lampreys). Placodermi (“jawfish”) -EXTINCT. Chondrichthyes (sharks and rays). Osteichthyes (“bony fish”). Amphibia. Vertebrates. Reptilia. Aves (birds). Mammalia. Phylum Chordata.

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Presentation Transcript
slide1

Phylum Chordata

Cephalochordata

(“lancelets”)

Urochordata

(“tunicates)

Agnatha

(jawless fish, e.g. lampreys)

Placodermi (“jawfish”) -EXTINCT

Chondrichthyes

(sharks and rays)

Osteichthyes (“bony fish”)

Amphibia

Vertebrates

Reptilia

Aves (birds)

Mammalia

slide2

Phylum Chordata

Cephalochordata

(“lancelets”)

Urochordata

(“tunicates)

Agnatha

(jawless fish, e.g. lampreys)

Placodermi (“jawfish”) -EXTINCT

Chondrichthyes

(sharks and rays)

Osteichthyes (“bony fish”)

Amphibia

Vertebrates

Reptilia

Aves (birds)

Mammalia

slide4

Marine Fish

Class Chondrichthyes

(“cartilaginous fish”)

Class Osteichthyes

(“bony fish”)

slide5

Bioactive Compounds from Osteichthyes:

Tetrodotoxin

Fugu spp.

(“Pufferfish”)

…and others.

Tetrodotoxin (TTX)

slide6

Voltage-Gated Sodium Channels

+ + + + + + + +

+ + - - - - - - - -

Voltage-Gated Ion Channels

K+

Na+

slide7

Action Potential

Action Potential

+ 50

Depolarizing

Phase

0

Repolarizing

Phase

Membrane Potential (mV)

- 50

Threshold

Resting Potential

- 70

Time (milliseconds)

slide8

Voltage-Gated Sodium Channels

+ + + + + + + +

- - - ++ -- - - - -

Action

Potential

K+

Na+

slide11

Bioactive Compounds from Chondrichthyes

Other Myths/Misconceptions?

“Sharks Don’t Have Immune Systems, But Don’t Get Infections”

slide14

Shark Liver Oil

Ingredients

71.9% Petrolatum

14% Mineral Oil

3% Shark Liver Oil

0.25% Phenylephrine HCl

slide15

Bioactive Compounds from Chondrichthyes: Squalamine

Stomach Tissue

(400 g)

Squalus acanthias

(“Dogfish Shark”)

Extract: 60% CH3CN/1% TFA

Extract Lipids: “Folch Method”

2:1 CHCl3/MeOH, Salt Wash

Aqueous Phase

Gel Filtration (Bio-Gel P-30)

C18 HPLC

SCX HPLC (Ion Exchange)

C4 HPLC

FAB-MS, 1H- and 13C-NMR, 2-D NMR (NOESY, HMQC)

Moore et al. (1993) Proc. Natl. Acad. Sci., 90: 1354-8.

slide16

Squalamine

MW 628.4739

Moore et al. (1993) Proc. Natl. Acad. Sci., 90: 1354-8.

slide18

Tissue Distribution of Squalamine

Amount

Tissue (µg/g tissue)

Stomach 1

Liver 4-7

Gallbladder 4-7

Spleen 2

Testes 2

Gills 0.5

Intestine 0.02

slide20

NHE1, 2 and 3 cDNA

(Transformed)

40 mM NH4Cl for 2-10 min.; Perfuse with Medium (Acidification)

2’, 7’-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-AM

(Intracellular pH Indicator)

Squalamine Inhibits Na+/H+ Exchanger (NHEs)

PS120 Fibroblasts

(No NHE)

Akhter et al. (1999) Am. J. Physiol., 276: 136-44.

slide21

Squalamine Inhibits H+ Efflux in NHE3-Transformed Cells

Akhter et al. (1999) Am. J. Physiol., 276: 136-44.

slide22

Squalamine Inhibition is Dose-Dependent

Akhter et al. (1999) Am. J. Physiol., 276: 136-44.

slide23

Squalamine Does NOT Inhibit H+ Efflux in NHE1- or NHE2-Transformed Cells

Akhter et al. (1999) Am. J. Physiol., 276: 136-44.

slide24

Squalamine Inhibition of NHE3-Transformed Cells Requires –COOH Terminal (76 Amino Acids)

Akhter et al. (1999) Am. J. Physiol., 276: 136-44.

slide25

Squalamine Inhibits Na+/H+ Exchange in Rabbit Illeal Brush Border Vesicles

22Na+ Uptake

Akhter et al. (1999) Am. J. Physiol., 276: 136-44.

slide30

MAPK Phosphorylation is pH Dependent

Stathopoulou et al. (2006) J. Exp. Biol., 209: 1344-54.

slide33

Pre-Clinical Trials with Squalamine

Xenograft Mice: Lung, Breast, Ovarian and Prostate Cancer

Allograft Rats: Brain and Breast Cancer

Inhibit Tumor Growth and Angiogenesis

Phase I/IIa Clinical Trials with Squalamine

Lung and Ovarian Cancer

MTD = 384 mg/m2/day; 120-h Continuous Infusion

Co-Administration with Other Drugs

slide34

Co-Administration of Squalamine

Control

Cisplatin

+ Squalamine

Cisplatin + Squalamine

Schiller and Bittner (1999) Clin. Cancer Res., 5: 4287-94

slide35

Squalamine and Macular Degeneration

Squalamine Lactate (MSI-1256F)

EVIZON™

(Genaera® Pharmaceuticals)