Background: A negative fluid balance during bed rest (BR) is accompanied by decreased plasma volume (PV) which contributes to cardiovascular deconditioning. Hypothesis: We hypothesized that increasing dietary sodium while controlling fluid intake would increase plasma osmolality (POSM), stimulate fluid conserving hormones, and reduce fluid/electrolyte (F/E) losses during BR; conversely, decreasing dietary sodium would decrease POSM, suppress fluid conserving hormones, and increase F/E losses. Methods: We controlled fluid intake (30 ml · kg-1 · d-1) in 17 men who consumed either a 4.0 ± 0.06 g · d-1 (174 mmol · d-1) (CONT; n = 6), 1.0 ± 0.02 g · d-1 (43 mmol · d-1) (LS; n = 6), or 10.0 ± 0.04 g · d-1 (430 mmol · d-1) (HS; n = 5) sodium diet before, during, and after 21 d of 6° head-down BR. PV, total body water, urine volume and osmolality, POSM, and F/E controlling hormone concentrations were measured. Results: In HS subjects, plasma renin activity (-92%), plasma/urinary aldosterone (-59%; -64%), and PV (-15.0%; 6.0 ml · kg-1; p < 0.05) decreased while plasma atrial natriuretic peptide (+34%) and urine antidiuretic hormone (+24%) increased during BR (p < 0.05) compared with CONT. In LS, plasma renin activity (+166%), plasma aldosterone (+167%), plasma antidiuretic hormone (+19%), and urinary aldosterone (+335%) increased with no change in PV compared with CONT (p < 0.05). Total body water did not change in any of the subjects. Conclusions: Contrary to our hypothesis, increasing dietary sodium while controlling fluid intake during BR resulted in a greater loss of PV compared with the CONT subjects. Reducing dietary sodium while controlling fluid intake did not alter the PV response during BR compared with CONT subjects.