CUBE Nerul, Navi Mumbai Home Locality Lab
Moina Model System
20 April 2020
Research Question: How does the low-oxygen condition (hypoxia) change the colour of the Moinas?
Short-term Objective: To prepare a Red-Moina Setup at home during the lockdown period.
Long-term Objective: To prove Histone modification due to low oxygen in Moina using Valproic Acid.
To some, science at home during the lockdown period seems impossible!
But for me, it is not so difficult to maintain my Moina culture atleast.
So, planning to prepare a Red-Moina Setup!
Design of the Red-Moina Set-up
8 Plastic Cups, Milk, ~2L Dechlorinated Water, Cloth pieces, dropper, Moinas!
Thought of taking 8 cups.
4 for control and 4 for the test.
200mL Dechlorinated Water + ~10 colourless Moinas + 1 drop of milk daily.
4 Controls so that each would be a replicate of one another.
200mL Dechlorinated Water + ~10 colourless Moinas + 6 drops of milk (not daily but on the alternate days till their number increases. If I add 6 drops of milk in 200mL of DC Water daily and as their number would be less, the water will become turbid, more bacterial growth will be there, very less oxygen would also be there as the bacteria will multiply. So, less Moinas won’t be able to survive. So till the 4th day when their number will become ~50-60, I will add 6 drops of milk daily.)
Here too, 4 test cups will be replicates of one another.
Half a bucket of tap water kept open for a day, we will get DC Water!
As the chlorine would evaporate, that Water would be Dechlorinated the next day!
Why 200mL Dechlorinated Water?
The cups are of more or less 200mL in volume.
Although the method which I and other CUBists follow is 250mL of DC Water + 1 and 6 drop(s) of milk, I will use 200mL of DC Water + 1 and 6 drop(s) of milk for maintaining Normal Oxygen Concentration or Normoxia and Low Oxygen Concentration or hypoxia respectively.
Why am I doing this?
I thought of giving the Moinas some work and see that how does it go in my home lab.
Earlier, I have tried this in CUBE Elphinstone, Mumbai as well as CUBE HBCSE, Mumbai
What will I prove after this experiment?
I will prove that the Red Moinas adapt to low oxygen condition (hypoxia-provided by me by adding more milk) by turning into Red Moinas.
This Red Colour would be due to the extra production oxygen-carrying protein pigment Haemoglobin.
How is adding more milk inducing hypoxia?
Milk contains bacteria as well as Carbohydrates, proteins, fats, vitamins and minerals which will act as a nutrient source for the bacteria.
We should note that the Dechlorinated Water which I will be using will also have some bacteria which will feed on the milk (as their nutrient source) and replicate.
So Milk bacteria + Culture Medium Bacteria will consume more oxygen as compared to the Moinas due to which less oxygen would be remaining for the Moinas to consume.
And now, to consume this less oxygen, the Moinas will start synthesizing the extra-haemoglobin through its four globin genes due to which they will appear red.
Because I am adding 6 drops of milk alternatively/daily, the hypoxia would be maintained throughout.
According to a 2004 Research Paper on Hypoxia-induced Synthesis of Hemoglobin in the Crustacean Daphnia magna Is Hypoxia-inducible Factor-dependent by Thomas Gorr and team in Journal of Biological Chemistry, it says that there are 4 globin genes present in Daphnia magna (a relative of Moina, that is why we are assuming the same in Moina too).
All four will be actively synthesising Haemoglobin in Hypoxia and in Normoxia, only a single gene would be active.
The quote from the Paper
“Protein products of the hb3 and hb2 genes are strongly induced by hypoxia, whereas the up-regulation of the hb1 gene is only moderate…”
Which to me, it means that only a single gene would be active in Normoxia!
This in-turn relates this to us to the 2019-Nobel Prize-winning Work by William G. Kaelin, Jr., Peter J. Ratcliffe and Gregg L. Semenza for their discoveries of hypoxia and how cells sense and adapt to oxygen availability.
You can correct me if I am wrong (I should be😬)!!
The link for the Paper:
My expectations at the end of the 7days or a week?
The 4 Control cups having 200mL of DC Water + 1 drop of milk daily + ~10 colourless Moinas (initially) will have ~60-70 Moinas with all the Moinas being colourless.
The 4 Test cups having 200mL of DC Water + 6 drop(s) of milk alternatively/daily + ~10 colourless Moinas (initially) should have ~100-120 Moinas in total out of which 60-70 should be orangish-red/red and remaining would be colourless/yellow.
Expectations for 21 April 2020?
Control as well as Test: ~20 Colourless Moinas in all the cups.
All these expectations are on the basis of the data which I got previously.
Photos, as well as Videos along with the updates, will be posted!