I would like to examine effects of particular drugs on D1 and D2 receptors using culture, but wondered if this is possible as people tend to look more at TH-positive cells. Thanks!
Hello
I not exactly sure what aspect of D1 and D2 receptors you are interested in. For example, protein expression levels, mRNA expression, cellular location etc, or simply to mark particular types of cell. It sounds like you are interested in labelling D1 and/or D2 expressing cells, perhaps with a goal of quantifying how many there are, or some change in distribution or cell morphology.
There are plenty of companies selling anti D1 or D2 antibodies that may work for immunohistochemistry and I have seen publications looking at least D1 expression in rat cultures from the striatum (PMID: 11320256). My experience of performing this type of staining is to just try it as antibodies can be hit or miss. Unfortunately, they are expensive for just a pilot and you may try asking the company for a sample. Also, on this issue I find that staining in cultures can be particularly error-prone with regard to achieving a genuine signal as there is not as much background material to compare to. With this in mind, some kind of control is a very good idea, such as antibody blocking, or even better, testing the antibody in tissue from D1/D2 knockout mice.
Aside from immunohistochemistry, there are many other ways you can look in on D1 and D2 depending on your question, e.g. Western blot, RT-PCR, receptor binding etc. There are also mice with fluorescence-tagged D1 and D2 receptors (e.g. PMID: 28436559), though clearly introducing them into your study may be a much more involved and costly affair. That said they deliver an excellent degree of confidence in the legitimacy of the expression.
Measuring expression of TH is as you say common. I would expect it to be far easier than D1 and D2 by immunohistochemistry, but it is quite a different but related question. Again, I am not sure what your goal is, but measuring TH is commonly used for identifying dopamine neurones, but do remember that TH is also involved in the production of norepinephrine and epinephrine so depending on from where you are taking the cultures, you may need to consider this.
Good luck!
Niall
This is very similar to HIV positive case transmission and its impact on rich proteins measuring is indefinite. Now there is no such mechanism to measure it but future surely holds the key
If your interesting in gene expression and if the drug will cause any effect the answer will be yes it is possible
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