Tuesday, July 17, 2018

Selective C1 Lesioning Slightly Decreases Angiotensin II Type I Receptor Expression in the Rat Rostral Ventrolateral Medulla (RVLM)


Erick A. Bourassa, Kristen A. Stedenfeld, Alan F. Sved, Robert C. Speth
Neurochemical Research Vol. 40, Issue 10 (October 2015)

The rostral ventrolateral medulla (RVLM) contains bulbospinal neurons that mediate sympathetic tone. These bulbospinal neurons are activated in two ways: baroreflex (drop in blood pressure) or by excitatory inputs from the paraventricular nucleus of the hypothalamus (PVN). The renin-angiotensin (RAS) system also plays a role in the regulation of peripheral blood pressure. Data suggests that elevated activity of the brain-specific renin-angiotensin system contributes to hypertension.
           
Within the RVLM there are two distinct types of neurons: C1 and non-C1. C1 Both C1 and non-C1 neurons are spinally projecting (bulbospinal), but they differ in that C1 neurons contain the enzymes required to synthesize epinephrine (tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT)), while non-C1 neurons do not. However, non-C1 neurons still express vGlu2, suggesting that they are glutamatergic like the C1 neurons. Studies have shown that both groups of neurons are involved in cardiovascular responses, but it is unknown whether the affects of angiotensin II (Ang II) in the RVLM are due to activation of the C1 neurons, the non-C1 neurons, or both. In this study, the AT1 receptor (Ang II receptor in the RVLM) is used to determine how active the C1s are versus the non-C1s in response to Ang II.

To test this, rats were either injected with DSAP (anti-dopamine-beta-hydroxylase-saporin) or MZAB (control) into one side of the RVLM. The other side of the RVLM was left alone to provide something to compare the experimental side of the RVLM to. The DSAP targets and destroys C1 neurons, while the MZAB has no effect. The rats were then decapitated 3-4 weeks post-injection and their brain was sectioned and stained for TH (indicative of C1 neurons). The number of C1 neurons in the RVLM of each rat was determined by examination of the processed brain tissue via light microscopy. The number of AT1 receptors was also determined, but by a different technique called quantitative angiotensin receptor autoradiography. It is important to note that two comparisons were made to determine results: comparison between right and left RVLM (contralateral) of the same rat to determine percent change, followed by comparison of the determined percent change between experimental RVLM and control RVLM of different rats.

DSAP produced a 57% depletion of C1 neurons, using the contralateral RVLM comparison, when compared to the contralateral comparison of the control MZAB injected RVLM. Also, the AT1 receptor binding was 10% lower in the experimental group when using the contralateral comparison, and 19% lower in the experimental group when comparing the binding between the DSAP and MZAB groups, not comparing contralaterally. If C1 neurons contained most of the AT1 receptors, as was previously thought, then a 57% decrease in C1 neurons would result in a similar decrease in the amount of AT1 receptors, but this was not the case. These results suggest that the non-C1 neurons in the RVLM contain most of the AT1 receptors.


The result of this study is significant to my work in the lab because it reminds us that while we focus on one role of the RVLM, it has other functions as well. It can be activated and inhibited by other pathways and it is important to remember that these other pathways may affect the results of our studies

- Ben Huber

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