Summaries of Articles/Lab Planning

Differential HSP70 Gene Expression: This article focused on looking at the HSP70 Gene expression in mussels by exposing the mussels to high temperatures and to different metals and solutions. The HSP70 is a heat shock protein that is synthesized in response to high heat. For example, in exposure to Mercury, transcription increased and slowly The results compared the different types of expressions – inducible or constitutive – and though more research is to be done, it can be supported that the general recovery time varies while the expression of these genes – HSC70 and HSP70 – are responsive to the length of exposure. This was interesting to learn about the genes responsible in protecting the mussels and specifically, the role minerals have on the protein synthesis.

Identification of differentially expressed genes of the Pacific oyster: Unlike the previous article I read, this article looks into the different genes expressed by the Pacific Oyster after exposure to prolonged temperature stress. With a control group exposed to a certain temperature, other oysters were exposed to higher temperatures to observe changes in their gene expressions over a 24 hour period.

The results detailed the relative expressions of different types proteins – including enzymes – between the control and treated groups, and also showed insight into the expression patterns in gills and mantles of the oysters after 3 and 7 day periods at 25 degrees versus 13 degrees. This included the cellular stress experienced 3, 7, 14, etc. days after the tissue was exposed to high heat. The question was how gene expression differed days after – whether there was down or up-regulation, along with whether greatly significant or not to an extreme degree. This brought insight to how oysters change their chemical makeup through transcription to respond to environmental changes in ensuring their survival.

Similar to the previous article, it was found that inducible forms of HSP70 (the heat shock protein) are expressed after exposure to higher heat than the constitutive form. It was found that the responses to the heat, there is also an internal cascade of protein synthesis within the oysters.

Lab Planning:

So far (weeks 1 & 2), I have been working with Professor Roberts to familiarize myself with the lab, and getting the logistics of the lab notebook, slack, WordPress, the lab calendar and lab safety. This past week, I have spent time reading through the articles and learning of the different discoveries already progressing in learning of mussel gene expression. I am currently finishing up last minute thoughts on the articles I have reviewed.

In this week, I am going to finish writing up the research plan by elaborating on the specifics as to my work being done in the coming weeks. I am going to be purchasing the mussels from a market or grocery store to bring into lab on Thursday to get them situated in the water. I am planning on buying about 45 mussels, and from there set up the mussels in a system. I will then read about sample collection and write up the sample collection plan in my notebook, with help from the Professor.

In the fourth week (next week), we will chose how we will work with the environmental stressor – temperature – to see the changes in gene expression and learn about their response mechanism. I will then. The main idea will be to change/vary temperature of the system the mussels are in to reflect the environmental changes they are already exposed to in their natural environment. We will then sample the mussels after 24 hours to reflect any changes. Within this period of time, there will be a control group that will not be exposed to these conditions to compare and contrast any changes in gene expression in different temperature conditions.

In week 5, we will get more hands on by completing our sampling. This will include RNA extraction, and plan out cDNA synthesis protocol. I have no experience in this part of the lab work, but I will be getting some guidance from the Professor and from other works for reference as to what I should do and how.

In week 6, I will work on finishing the RNA extractions, perform a cDNA synthesis and plan out a qPCR protocol.

Week 7 through 9 will focus on the writing aspect of research after we have completed running the qPCR and establishing the initial findings. I will learn how to and complete writing an Abstract, tie up and last analysis from the experiments and read through relevant papers that have undergone similar experiences. I will then start and complete my poster, practice my presentation and present my findings before other fellow scientists to summarize my work in the lab.