THE MOLECULAR NEUROBIOLOGY LABORATORY
The Methods
Molecular Biology
Heart rate variability and sympathetic nerve recordings
Immunohistochemistry
We use a number of different ways to characterise nerve cell types. This includes
(i) immunohistochemistry, using double labelling techniques and confocal microscopy;
(ii) gene expression using standard RT-PCR and Real Time PCR for quantitative
analysis, and (iii) heart rate variability and sympathetic nerve recordings.
Molecular Biology
Using mRNA analysis as a key to what
is going on inside the cell, we look at the expression of genes for
the different neuroreceptors and transmitters, using RNA extraction
and reverse transcription-polymerase chain reaction (RT-PCR) as our
main approach. We also use quantitative real time PCR to define the
relative expression levels of the different genes and can determine
mRNA expression in single cells. This approach helps us overcome the
problems associated with cellular heterogeneity, even within a small
defined region of the brain. |
|
|
| Real-time amplification of PCR products
from the brainstem |
Gel analysis of PCR product looking at
NMDA receptor splice variants |
|
TOP
Heart rate variability and sympathetic nerve recordings
 |
The Autonomic and Sensory Neurobiology Laboratory is one of the first research groups worldwide to take advantage of a new wireless technology for measuring blood pressure and sympathetic nerve activity. We are using this to look at short and long term control of blood pressure in our study of hypertension and kidney disease. One way we will analyse this data is to determine heart rate variability – as a measure of autonomic control of the heart.
|
|
LabChart recording of data recorded from a rat using a
Telemetry Research Transmitter and Receiver.
|
TOP
Immunohistochemistry
| |
Immunohistochemistry is a
powerful tool that can be used to determine what proteins are expressed
in different nerve cells, and importantly, allows us to combine anatomy
with phenotype to a high degree of detail. We use immunohistochemistry
in conjunction with fluorescent labelling and a confocal laser scanning
microscope, resulting in the collection of some very impressive data,
not to mention beautiful pictures. In the nervous system we can also take
advantage of the anterograde and retrograde transport of molecules within
the nerve fibre, allowing us to determine connection pathways between
different clusters of nerve cells. |
 |
|
Confocal image of cells in the RVLM region labelled with
tyrosine hydroxylase and retrograde tracer. |
TOP
|
![[ Murdoch University logo and link to homepage ]](/images/header/cs/logo/mu_logo_lhs_45.gif){kind=logo}
