Keiko Motonaga

Identification of Three Isoforms for the Na+-dependent Phosphate Cotransporter (NaPi-2) in Rat Kidney

We have isolated three unique NaPi-2-related protein cDNAs (NaPi-2α, NaPi-2β, and NaPi-2γ) from a rat kidney library. NaPi-2α cDNA encodes 337 amino acids which have high homology to the N-terminal half of NaPi-2 containing 3 transmembrane domains. NaPi-2β encodes 327 amino acids which are identical to the N-terminal region of NaPi-2 containing 4 transmembrane domains, whereas the 146 amino acids in the C-terminal region are completely different. In contrast, NaPi-2γ encodes 268 amino acids which are identical to the C-terminal half of NaPi-2. An analysis of phage and cosmid clones indicated that the three related proteins were produced by alternative splicing in the NaPi-2 gene. In a rabbit reticulocyte lysate system, NaPi-2 α, β, and γ were found to be 36, 36, and 29 kDa amino acid polypeptides, respectively. NaPi-2α and NaPi-2γ were glycosylated and revealed to be 45- and 35-kDa proteins, respectively. In isolated brush-border membrane vesicles, an N-terminal antibody was reacted with 45- and 40-kDa, and a C-terminal antibody was reacted with 37-kDa protein. The sizes of these proteins corresponded to those in glycosylated forms. A functional analysis demonstrated that NaPi-2γ and -2α markedly inhibited NaPi-2 activity in Xenopus oocytes. The results suggest that these short isoforms may function as a dominant negative inhibitor of the full-length transporter.

Energy Requirement Assessment and Water Turnover in Japanese College Wrestlers Using the Doubly Labeled Water Method

Estimated energy requirements (EERs) are important for sports based on body weight classifications to aid in weight management. The basis for establishing EERs varies and includes self-reported energy intake (EI), predicted energy expenditure, and measured daily energy expenditure. Currently, however, no studies have been performed with male wrestlers using the highly accurate and precise doubly labeled water (DLW) method to estimate energy and fluid requirement. The primary aim of this study was to compare total energy expenditure (TEE), self-reported EI, and the difference in collegiate wrestlers during a normal training period using the DLW method. The secondary aims were to measure the water turnover and the physical activity level (PAL) of the athletes, and to examine the accuracy of two currently used equations to predict EER. Ten healthy males (age, 20.4±0.5 y) belonging to the East-Japan college league participated in this study. TEE was measured using the DLW method, and EI was assessed with self-reported dietary records for ~1 wk. There was a significant difference between TEE (17.9±2.5 MJ•d-1 [4,283±590 kcal•d-1]) and self-reported EI (14.4±3.3 MJ•d-1 [3,446±799 kcal•d-1]), a difference of 19%. The water turnover was 4.61±0.73 L•d-1. The measured PAL (2.6±0.3) was higher than two predicted values during the training season and thus the two EER prediction equations produced underestimated values relative to DLW. We found that previous EERs were underestimating requirements in collegiate wrestlers and that those estimates should be revised.

Energy Deficit Required for Rapid Weight Loss in Elite Collegiate Wrestlers

To determine energy density for rapid weight loss (RWL) of weight-classified sports, eight male elite wrestlers were instructed to lose 6% of body mass (BM) within 53 h. Energy deficit during the RWL was calculated by subtracting total energy expenditure (TEE) determined using the doubly labeled water method (DLW) from energy intake (EI) assessed with diet records. It was also estimated from body composition change estimated with the four-component model (4C) and other conventional methods. BM decreased significantly by 4.7 ± 0.5 kg (6.4 ± 0.5%). Total body water loss was the major component of the BM loss (71.0 ± 7.6%). TEE was 9446 ± 1422 kcal, and EI was 2366 ± 1184 kcal during the RWL of 53-h; therefore, the energy deficit was 7080 ± 1525 kcal. Thus, energy density was 1507 ± 279 kcal/kg ∆BM during the RWL, comparable with values obtained using the 4C, three-component model, dual energy X-ray absorptiometry, and stable isotope dilution. Energy density for RWL of wrestlers is lower than that commonly used (7400 or 7700 kcal/kg ΔBM). Although RWL is not recommended, we propose that commonly practiced extreme energy restriction such as 7400 or 7700 kcal/kg ΔBM during RWL appears to be meaningless.

Muscle glycogen depletion does not alter segmental extracellular and intracellular water distribution measured using bioimpedance spectroscopy

Although each gram of glycogen is well known to bind 2.7-4.0 g of water, no studies have been conducted on the effect of muscle glycogen depletion on body water distribution. We investigated changes in extracellular and intracellular water (ECW and ICW) distribution in each body segment in muscle glycogen-depletion and glycogen-recovery condition using segmental bioimpedance spectroscopy technique (BIS). Twelve male subjects consumed 7.0 g/kg body mass of indigestible (glycogen-depleted group) or digestible (glycogen-recovered group) carbohydrate for 24 h after a glycogen-depletion cycling exercise. Muscle glycogen content using 13C-magnetic resonance spectroscopy, blood hydration status, body composition, and ECW and ICW content of the arm, trunk, and leg using BIS were measured. Muscle glycogen content at the thigh muscles decreased immediately after exercise (glycogen-depleted group, 71.6u2009±u200912.1 to 25.5u2009±u200910.1 mmol/kg wet wt; glycogen-recovered group, 76.2u2009±u200916.4 to 28.1u2009±u200916.8 mmol/kg wet wt) and recovered in the glycogen-recovered group (72.7u2009±u200921.2 mmol/kg wet wt) but not in the glycogen-depleted group (33.2u2009±u200912.6 mmol/kg wet wt) 24 h postexercise. Fat-free mass decreased in the glycogen-depleted group ( P < 0.05) but not in the glycogen-recovered group 24 h postexercise. However, no changes were observed in ECW and ICW content at the leg in both groups. Our results suggested that glycogen depletion per se does not alter body water distribution as estimated via BIS. This information is valuable in assessing body composition using BIS in athletes who show variable glycogen status during training and recovery. NEW & NOTEWORTHY Segmental bioimpedance spectroscopy analysis reveals the effect of muscle glycogen depletion on body segmental water distribution in controlled conditions. Despite the significant difference in the muscle glycogen levels at the leg, no difference was observed in body resistance and the corresponding water content of the extracellular and intracellular compartments.