User:Enabulei/Firefly squid/Bibliography

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 * Hamanaka, T., Michinomae, M., Seidou, M., Miura, K., Inoue, K., & Kito, Y. (2011). “Luciferase Activity of the Intracellular Microcrystal of the Firefly Squid, Watasenia Scintillans.” FEBS Letters, 585(17), 2735-2738. doi:10.1016/j.febslet.2011.07.033.
 * Firefly squid (Watasenia scintillans) is a readily available deep-sea organism for research purposes because it comes to the shallow waters in the Sea of Japan to spawn each Spring and is easily captured in fisherman nets.
 * W. scintillans is bioluminescent with multiple light producing organs located around each eye and at the tips of the fourth pair of ventral legs. The biochemical details of its bioluminescence are still not known. Within these light producing organs, there are numerous rod-like bodies which have a crystalline structure.
 * This study analyzed these rod-like structures with several different imaging techniques, including electron microscopy and optical microscopy. The researchers used the X-ray powder diffusion method to demonstrate that these rods were tightly packed microcrystals. After isolating these microcrystals, they discovered that in the presence of ATP and Mg2+, the intense blue light of bioluminescence was emitted from these microcrystals.
 * Using detergents to remove any membrane bound proteins, they found no change in the bioluminescence produced by these light organs, contradicting the previously reported theory that the luciferase needed by these squid was membrane bound.
 * These researchers concluded that these microcrystals contained both the luciferin and the luciferase used by the Firefly squid to produce its bioluminescence and that the reaction occurred within the crystals.


 * Teranishi, K., & Shimomura, O. (2008). “Bioluminescence of the Arm Light Organs of the Luminous Squid Watasenia Scintillans.” Biochimica et Biophysica Acta (BBA) - General Subjects, 1780(5), 784–792. doi:10.1016/j.bbagen.2008.01.016
 * This team of scientists studied the biochemical mechanism of bioluminescence in the Firefly squid (Watasenia scintillans). Their work centered on the light producing organs on the ventral legs.
 * The luciferin reactant of the bioluminescence reaction had already been isolated and identified but the luciferase enzyme has not been conclusively isolated or studied.
 * Their attempts to isolate the luciferase from these light emitting organs proved very difficult until they used high concentration sucrose solutions to extract, stabilize, and isolate the active component.
 * They found the luciferase activity was isolated in particulate form though they could not isolate it further.
 * They hypothesized that it was part of a membrane bound particle.


 * Tsuji, F. I. (2005). “Role of Molecular Oxygen in the Bioluminescence of the Firefly Squid,Watasenia Scintillans.” Biochemical and Biophysical Research Communications, 338(1), 250–253. doi:10.1016/j.bbrc.2005.08.215.
 * This study focused on identifying the essential reactants of the bioluminescence reaction in the Firefly squid.
 * It had previously been established that it was an ATP requiring reaction.
 * They created a homogenate from the arm organs of the squid and altered the composition of the homogenate to see what was needed for the bioluminescence reaction to occur.
 * They found that molecular oxygen was a requirement because when air/oxygen was replaced by pure hydrogen gas or carbon dioxide, the reaction stopped and restarted when air was reintroduced.
 * They tested other chemicals and found that Mg2+ was also a requirement. Alkaline pH (between 7.50 to 9.50) with optimal reaction occurring at 8.80 also affected the magnitude of the reaction.


 * Sato, N., Tsuda, S., Alam, N.E., Sasanami, T., Iwata, Y., Kusama, S., Inamura, O., Yoshida, M., & Hirohashi, N. (2020). “Rare Polyandry and Common Monogamy in the Firefly Squid, Watasenia Scintillans.” Scientific Reports, 10(1), 10962. doi:10.1038/s41598-020-68006-1.
 * Cephalopods species are presumed to have a polyandry mating system – a female will copulate with more than one male for increased fitness of having one of several different gene pools for each of her offspring.
 * Male cephalopods package their sperm in spermatangia which are then deposited in bilateral specialized pouches in the female’s neck collar.
 * Sampling female cephalopods from other species, the female typically has unequal numbers of spermatangia on each side and genotyping of these sperm deposits reveals that they come from different male mates.
 * The female Firefly squid almost always had equal numbers of spermatangia in each pouch.  When the sperm in these pouches of female Firefly squid was genotyped, in 95% (18/19) of females only sperm from one male was found and all of the offspring in a clutch had the same father.


 * Gimenez, G., Metcalf, P., Paterson, N.G., & Sharpe, M. L. (2016). “Mass Spectrometry Analysis and Transcriptome Sequencing Reveal Glowing Squid Crystal Proteins Are in the Same Superfamily as Firefly Luciferase.” Scientific Reports, 6(1), 27368. doi:10.1038/srep27638.
 * This is another study trying to further outline the molecular basis of bioluminescence of the Firefly squid since it seems different than that of other species.
 * The scientists used high-throughput mRNA transcripts of extracts from the arm tip tissue and the mantle tissue, where the arm and cutaneous photophores are located.
 * The microcrystals dissolved quickly during extraction but could be stabilized with a high sucrose solution and stored at 4°C.
 * They isolated three protein bands from the arm tissue samples and found a single homologous protein in the cutaneous samples.
 * None of these four proteins were similar to other known lucriferases so their molecular mechanism is still not understood.
 * Given the different bioluminescent properties of the arm tip (very intense blue light) versus the cutaneous (low intensity blue or green light) photophores, one hypothesis was that the three proteins found in the microcrystals of the arm tip formed a complex or otherwise worked together to generate a different quality of bioluminescence than the single protein of the cutaneous photophore.


 * Takeuchi, H., Morita, R., Shirai, Y., Nakagawa, Y., Terashima, T., Ushikubo, S., & Matsuo, T. (2014). “Lowering Effect of Firefly Squid Powder on Triacylglycerol Content and Glucose-6-Phosphate Dehydrogenase Activity in Rat Liver.” Journal of Oleo Science, 63(12), 1293–1301. doi:10.5650/jos.ess14149.
 * Firefly squid come to the surface between March and May and are easily caught with nets and are eaten. Unlike most squid species, especially the Japanese flying squid where the internal organs and head are removed before ingestion, the custom for eating Firefly squid is to ingest the entire organism.
 * Squid in general are known to be high in cholesterol content so people with elevated serum cholesterol levels may be hesitant to eat squid in their diet.
 * A previous study using edible Japanese flying squid found a decrease in liver and serum lipid levels in rats when flying squid was used to supplement their diet.
 * This study sought to evaluate whether dietary supplements with whole Firefly squid had an effect on lipids in rats. Using three groups of identical rats (control, flying squid supplement, and whole Firefly squid supplement) the researchers found that ingestion of either squid supplement resulted only in decreased liver lipid levels when compared to adult control rats.
 * Looking at DNA microarrays, they determined that the glucose-6-phosphate dehydrogenase gene expression was significantly reduced when compared to control rats and this may be part of the mechanism by which Firefly squid supplements can reduce hepatic lipid concentration.


 * Michinomae, M., Masuda, H., Seidou, M., & Kito, Y. (1994). “Structural Basis for Wavelength Discrimination in the Banked Retina of the Firefly Squid Watasenia Scintillans.” Journal of Experimental Biology, 193(1), 1–12. doi: 10.1242/jeb.193.1.1.
 * Firefly squid are unique among cephalopods in having three visual pigments in its retina, each with a unique spectral sensitivity, which may result in color vision.
 * The dorsal retina has an outer segment (OS) thickness of 200 μm and a thinner inner segment (IS) of 50 μm. Cells of the dorsal retinal inner segment contain one visual pigment with spectral absorbance maximum at 484 nm, similar to chromophore retinal A1.
 * The ventral retina is thicker with an outer segment of 600 μm and an inner segment of 200 μm.
 * Cells in the distal 2/3 of the dorsal retina OS contain a visual pigment with maximal absorbance at 470 nm, similar to chromophore retinal A4.
 * Cells in the proximal 1/3 of the dorsal retina OS contain a third visual pigment with maximum absorbance at 500 nm similar to retinal pigment A2.
 * The finding that each visual pigment resides in different retinal cells raises the possibility that the Firefly squid is unique among cephalopods in having color vision.


 * Kröger, R. H. H., & Gislén, A. (2004). “Compensation for Longitudinal Chromatic Aberration in the Eye of the Firefly Squid, Watasenia Scintillans.” Vision Research, 44(18), 2129–2134. doi:10.1016/j.visres.2004.04.004.
 * In order for an organism to have color vision, it must be able to focus different wavelengths of light at different distances from the lens, an ability called longitudinal spherical aberration (LSA).
 * Since the Firefly squid is unique among cephalopods in having three retinal pigments, color vision was possible if the squid was capable of LSA.
 * Surprisingly the squid did not have any LSA, the mechanism used by fish to have color vision.
 * The researchers did find that the banked structure of the Firefly squid with the separation of three retinal pigments at different positions within the retina, almost perfectly performed the same function as LSA, making it highly likely that they had color vision.
 * There were other optical benefits of having this banked retinal structure.

Enabulei
 Elizabdo 


 * Tsuji, F. I. (1985). ATP-dependent bioluminescence in the firefly squid, Watasenia scintillans. Proceedings of the National Academy of Sciences, 82(14), 4629-4632. https://doi.org/10.1073/pnas.82.14.4629
 * Tsuji collected firefly squid at the Toyama Bay and performed a series of experiments on them to determine the requirements for emission of bioluminescence in the squid.
 * Experiments were performed by homogenizing completely dark or weak luminescent arm organs
 * 1 M NaCl was injected into the homogenate, one with dithioerythritol and one without, both however did not trigger light emission (testing so see if certain solutions would trigger light emission)
 * A solution of ATP was injected into the homogenate and triggered light emission
 * A series of experiments were performed on the homogenate using ATP and compounds related, however, ATP showed the highest initial maximal light intensity compared to every other compound
 * ATP-dependent reaction was determined to have luciferin, ATP, MG2+, optimum pH of 8.8, as its main components
 * A conclusion was made that ATP was essential for the bioluminescent aspect of the firefly squid
 * Tsuji, F. I. (2002). Bioluminescence reaction catalyzed by membrane-bound luciferase in the “firefly squid,” Watasenia scintillans. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1564(1), 189-197. https://doi.org/10.1016/s0005-2736(02)00447-9
 * The experiments were carried out in Namerikawa at Toyama Prefectural Fisheries Research Institute
 * About 30-40 firefly squid were collected and kept in a tank
 * 15 live specimens were used to prepare a homogenate
 * 3 small black organs at the tip of each of the 4th pair of ventral arms was snipped off and homogenized where they slowly lost luminesce
 * A series of experiments were performed on those hemoginates
 * 2 mixtures were made, one with ATP, .001 M MgCl2+ a microliter of 1 M Tris-HCL, & the pH of 8.26, the other identical except for the ATP. The luciferase pellets were added to the second mixture(without ATP) where they found no light to be detected, thus luciferase was not activated by ATP
 * When luciferin and luciferase were exposed to Tris-HCL, pH 8.26, there was an increase in light intensity.
 * Final results
 * Bioluminescence requires molecular oxygen
 * ATP is required in the Watasenia reaction
 * It is unknown whether the luciferin, coelenterazine disulfate, is synthesized from the beginning or if it comes from the diet
 * The reaction is enhanced by a high alkaline pH of 8.8
 * Teranishi, K., & Shimomura, O. (2008). Bioluminescence of the arm light organs of the luminous squid Watasenia scintillans. Biochimica et Biophysica Acta (BBA) - General Subjects, 1780(5), 784-792. https://doi.org/10.1016/j.bbagen.2008.01.016
 * Firefly squid were captured offshore in Toyama Bay, Japan where the light organs would be picked off of the squid and homogenized for experimentation
 * Through centrifugation, a supernatant was made containing partially purified active substance that they names “arm-organ extract” and used for further experimentation
 * First, they wanted to find the pH that suited luciferase activity best, meaning when luciferase was most active and they found that to be 6.2
 * They found through trial and error that luciferase activity in hongenates and arm-organ extracts could be markedly stabilized by adding some sucrose
 * Also found that the concentrations of coelenterazine disulfate ( a luciferin), ATP and MgCl2 have strong effects on the light intensity of luminescence reaction catalyzed by the arm-organ extract
 * Final conclusions
 * Luciferase in the arm light organs was a kind of cellular particle
 * Using sucrose, luciferase can be extracted from the light organs and partially pureed, yielding particulate material with strong luciferase activity
 * The particulate luciferase catalyzes the luminescence reaction of coelenterazine disulfate in presence of ATP and Mg2+
 * Consistent failure in the attempts of solubilizing particulate luciferase and the very high specificity to coelenterazine disulfate led them to conclude the existence of an unusual type of luciferase active site in this luminescence system.
 * The active site might be formed only when luciferase molecule is bound with unidentified cellular particle possibly in the presence of ATP, or when bound at an interface area of two or more different molecules composing the particle, in a way that increases the specificity to the substrate coelenterazine disulfate
 * Sato, N., Tsuda, S., Alam, N. E., Sasanami, T., Iwata, Y., Kusama, S., Inamura, O., Yoshida, M., & Hirohashi, N. (2019). Polyandry is extremely rare in the firefly squid, Watasenia scintillans. https://doi.org/10.1101/2019.12.13.875062
 * Often cephalopod species reproduce polyamorously, however, it is suggested that some reproduce monogamously because searching for extra mates is costly and may interfere of their own resources
 * Firefly squid were obtained from fishery catches off of Oki island and Sakai-port by Toyama bay where they began their experiments with spawning
 * Male firefly squid are said to disappear from mid-february to mid-march, thus females carry a sperm sac to fertilize when necessary
 * After a female spawned, the females were transported immediately after the catch
 * A paternity test was performed on the females by removing their seminal receptacle and extracting DNA to perform a percentage DNA analysis for identification
 * After the paternity test was run, they found that 94.7% of females, or 18 out of the 19 collected, had spermatangia stored from a single male
 * Final results = most cephalopod females are known to usually behave “promiscuously” thus having multiple mates, however, the firefly squid females were found to be monogamous
 * Patel, K., & Pee, D. (2011). Watasenia scintillans. Animal Diversity Web. https://animaldiversity.org/accounts/Watasenia_scintillans/
 * The Watasenia scintillans, or firefly squid is a deep sea creature found in the ocean at depths of 200-400m found mainly in Japan
 * It is a small cephalopod, growing up to 8cm, approximately 3inches, in size with a lifespan of 1 year
 * The squid is primarily dark in color and has a bioluminescent component around the eyes, hooks and tentacles that can be used as a mating tactic, attraction for prey, attraction for a mate, and for general communication
 * The squid comes to the surface to mate during breeding season that lasts from March-May
 * During their breeding period, the firefly squids rise to the surface in huge schools along the shoreline
 * They will also come to the surface at night to capture prey as they are carnivorous