Konnichi wa from the Primate Research Institute – Week 5 (July 12-19)
One month at the PRI is officially in the books, and I must say everyday has been a great learning experience. Dr. Imamura and his research team have been extremely helpful every step of the way, and I feel very confident in my new laboratory skills which should translate nicely to the next month of research. This week, we kicked off our training wheels and jumped into our official RT-PCR experiments.
RT-PCR: Express Yourself!
This week, we were able to begin our first sets of experiments to address the question: “What genes involved in spermatogenesis are being expressed within male macaque monkey testis tissue?” Previously, our research team has extracted the RNA from the macaque samples, generated the protein-coding DNA sequences from said RNA, and designed a plethora of primers for the RT-PCR experiments. The end goal of these experiments will be the generation of a genetic profile that characterizes macaque spermatogenesis, which will benefit future research into genetic studies of human and non-human primate reproduction. Here’s a quick glimpse at some early RT-PCR data for β-actin, a gene that is expressed in nearly every tissue, thus making it a valuable “control gene” to make sure our samples are of good quality.
Here we have five wells, each with a different DNA sample. From left to right – DNA ladder of known values, a negative control with water, and three macaque samples. Our macaque samples express β-actin just like we expected; we were “actin” really happy when we saw this!
These results show the expression of SCP3, a protein that is crucial for proper chromosomal division during meiosis. From left to right – DNA ladder, negative control, and our three macaque samples: juvenile, adult, and adult. We expected brighter bands for the adults since SCP3 has been found in more adult cell types than those of juveniles, which is just what we see!
We will repeat these experiments using different combinations of genes and macaque samples, which will yield a very large dataset of what’s being expressed and, just as important, what’s not being expressed. This means many more long days in the laboratory and many more cups of coffee and tea. Alas, the things we do for data!
Simultaneously, I’m continuing the 2nd component of our research project involving immunohistochemistry with fellow labmates, Kuroki-san and Ito-san. This past week was focused primarily on RT-PCR, but we expect to start generating more data of stained tissue samples within the next week, as well. I’ll be the first to admit that the immunohistochemistry images are more visually captivating than fluorescent PCR bands, so I’ll try to squeeze in some images of stained cells in later blogposts!
Finding My Zenter
Thanks to Kaori-san, the administrator for the Molecular Biology section, I was able to partake in a traditional Buddhist ceremony called zazen (seated meditation). Early Sunday morning, we gathered at a Buddhist otera (temple) to begin our journey inwards. Kaori-san, her two small daughters, Ito-san, and I took our places, then our Buddhist instructor taught us the proper methods of sitting, breathing, and concentration. The calm of the morning, birds chirping in the nearby garden, and the soft smell of incense created the perfect environment for our meditation.
Ito-san and I found a great spot for zazen with a nice view of the outside garden.
Kaori-san, her friend, and their daughters all joined Ito-san and me for zazen. The small children really exaggerate my height!
If you feel like your mind is wandering too much, you can signal to the jikido (zen instructor) that you want to be struck on the shoulder with a kyosaku (wooden stick). No pain, no gain!
Down the Road
I have one more month left in Japan, and from the looks of it, it will be even more exciting! My trip to Kyushu University to give a special lecture is coming up next weekend, the research should continue humming along, and many more opportunities to experience Japanese culture with fellow PRI researchers surely await. I’m also looking forward to the historic ukai (cormorant fishing) here in Inuyama, which I mentioned last week. This is where fishermen use cormorants, a kind of sea bird, to dive in the water and catch fish for them. As always, thanks for keeping up with my research and experiences here in Japan! Until next week, matane!
This research is funded by the National Science Foundation East Asia and Pacific Summer Institutes for U.S. Graduate Students (EAPSI) in collaboration with the Japan Society for the Promotion of Science (JSPS).