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      Dissociation and Enrichment of Rat Atrial Myocytes Containing Atrial Natriuretic Factor (ANF)

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          Methodologies developed for the dissociation and subsequent enrichment of muscle and nonmuscle cells from atrial myocardium were used to evaluate the contribution of these cell populations to the natriuretic, diuretic and vasoactive properties of crude atrial tissue extracts. Suspensions of single cells, which contained approximately 34% myocytes, were prepared from atrial tissue blocks with a collagenase-trypsin digestion followed by gentle mechanical disruption. Differential centrifugation and unit gravity sedimentation techniques were employed to enrich the ‘muscle’ and ‘nonmuscle’ cell suspensions to a purity of approximately 91 and 95%, respectively. Cell extracts were bioassayed for natriuretic activity in saline-expanded, pentobarbital-anesthetized, female rats. Extracts obtained from ‘initial’ and ‘muscle’ cell suspensions significantly enhanced sodium and chloride excretion as well as urine flow while extracts from ‘nonmuscle’ cell suspensions had no effect on renal function. Sodium excretion was dose-dependent and increased linearly with increasing numbers of extracted and infused myocytes. This simple two-step centrifugation and sedimentation protocol can be utilized to obtain enriched atrial myocyte populations for subsequent physiologic and biochemical studies.

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          Author and article information

          Horm Res Paediatr
          Hormone Research in Paediatrics
          S. Karger AG
          28 November 2008
          : 24
          : 1
          : 26-37
          Department of Physiology and Biophysics, University of Cincinnati, College of Medicine, Cincinnati, Ohio, USA
          180536 Horm Res 1986;24:26–37
          © 1986 S. Karger AG, Basel

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          Page count
          Pages: 12


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