the establishment of a primary culture system of proximal tubule segments using specific markers from normal mouse kidneys建立的主要文化系统从正常老鼠肾脏近端小管部分使用特定标记.pdf

Int. J. Mol. Sci. 2012, 13, 5098-5111; doi:10.3390/ijm OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 /journal/ijms Article The Establishment of a Primary Culture System of Proximal Tubule Segments Using Specific Markers from Normal Mouse Kidneys Masumi Kamiyama, Michelle K. Garner, Kristina M. Farragut and Hiroyuki Kobori * Department of Physiology, Hypertension and Renal Center of Excellence, Tulane University Health Sciences Center, 1430 Tulane Avenue, LA 70112, USA; E-Mails: mkamiyam@ (M.K.); mgarner@ (M.K.G.); kfarragu@ (K.M.F.) * Author to whom correspondence should be addressed; E-Mail: hkobori@; Tel.: +1-504-988-2591; Fax: +1-504-988-0911. Received: 21 March 2012; in revised form: 5 April 2012 / Accepted: 18 April 2012 / Published: 23 April 2012 Abstract: The proximal tubule contains the highest expression of angiotensinogen mRNA and protein within the kidney and plays a vital role in the renal renin-angiotensin system. To study the regulation of angiotensinogen expression in the kidney in more detail, the proximal tubule needs to be accurately isolated from the rest of the nephron and separated into its three segments. The purpose of this study was to design a novel protocol using specific markers for the separation of proximal tubule cells into the three proximal tubule segments and to determine angiotensinogen expression in each segment. Kidneys were removed from