단련한 흰쥐의 속근 및 지근에 Glycogen 과저장이 운동 지구력에 미치는 영향

연동수(Yeoun, Dong-Soo),황수관(Hwang, Soo-Kwan),김인숙(Kim, In-Sook),고성경(Ko, Seong-Kyeong),남택상(Nam, Taick-Sang),강두희(Kang, Doo-Hee) 대한생리학회 1988 대한생리학회지 Vol.22 No.1

This study was carried out to investigate relationships between maximal running time (MRT) and glycogen supercompensation in fast twitch white fibers (white vastus, WV), fast twitch red fibers (red vastus, RV) and slow twitch red fibers (soleus muscle, SM) of endurance-trained rats. Male rats of a Sprague-Dawley strain were divided into the trained groups and untrained groups. Untrained groups were acquired to run on the treadmill 10 minutes for 3 days and remained rest and maintained with mixed diet for 4 weeks. For last 10 days of resting period, the untrained rats were divided into 3 groups i.e. mixed diet (untrained control), high and low carbohydrate (CHO) diet groups. And each group was subdivided into 2 groups, one group was tested for the MRT and the other was sacrificed to measure the blood glucose, blood lactate, glycogen contents of liver and muscles. The experimental groups were trained on treadmill by a modified method of Constable et al. (1984) maintained with mixed diet for 4 weeks. After measurement of MRT of this group, they were also divided into high and low CHO groups and fed with these diet for 2 days and MRT of each group was measured again to see the effect of high or low CHO feeding on the MRT. Each group was maintained with the same diet for next 2 days during which some of the rats were sacrificed at given time intervals for the measurements of blood glucose and lactate, liver and the muscles glycogen. The results were summarized as follows; 1) In the untrained group, there were no significant differences between subgroups in MRT, glycogen conent of SM, RV and WV. But blood glucose concentration and glycogen content of liver of low CHO group were significantly lower than those of mixed diet group. 2) The MRT and glycogen content of SM, RV and WV of trained mixed diet group were significantly increased compared to those of untrained mixed diet group, but there was no significant difference in glycogen content of liver. 3) MRT of trained mixed, high CHO and low CHO groups were 137±9.8, 176±9.8 and 129±7.3 min respectively with the significant difference between them. 4) There were no differences in blood lactate concentrations between the trained high and low CHO groups immediately after maximal running and during recovery period. 5) Glycogen contents in RV and SM of trained high CHO group were significantly increased, and glycogen contents in RV, WV and liver of trained low CHO group were significantly decreased compared to those of trained mixed diet group. 6) Immediately after maximal running, the blood glucose concentrations of trained high CHO and low CHO groups were 73±4.0 and 67±6.9mg% respecitively. The blood glucose of the trained high CHO group was fully recovered within one hour by feeding. But blood glucose concentration of low CHO group was slowly recovered up to 114±4.1mg% after two hours of feeding and maintained. Those values were still significantly lower than that of trained mixed diet group. The synthetic rates of glycogen in liver and muscles during the recovery period followed the similar time course of the blood glucose recoveries in each group. These results suggest that an increase in MRT of trained high CHO group was attributed to the glycogen supercompensation in slow twitch muscle fibers. And a decrease in MRT of trained low CHO may be due to decreased glycogen contents of liver and muscles. The results also suggest that glycogen supercompensation was more evident in slow twitch red fibers of endurance-trained rats and blood glucose is one of the limiting factors of glycogen synthesis.

운동부하 흰쥐의 하지골격근에서 과축적 당원의 이용양상

전춘배,안종철,송대흡,이석강 영남대학교 의과대학 1997 Yeungnam University Journal of Medicine Vol.14 No.1

골격근의 과축적 당원의 운동 중 이용 양상과 골격근의 구성 섬유 특성에 따른 당원의 이용 양상을 규명한 본 연구 결과를 요약하면 다음과 같다. 정상 군의 당원 농도는 가자미근에서 4.57±0.34 mg/gm wet wt.(이하 동일 단위)였으며 적색 비복근은 5.11±0.24, 족척근은 6.55±0.20으로서 족척근이 가장 높았다. 가자미근의 당원 농도는 운동 부하 후 당투여 1시간 군에서 정상 군에 비해서 약 1.9배 높았으며 적색 비복근은 1.8배 높았다. 족척근의 당원 농도는 운동 후 당 투여 군에서 정상 군과 통계적으로 유의한 차이가 없었다. 간 당원 농도는 정상군에서 41.0±1.47이였으며 대조 군에서는 정상 군의 2.9%에 불과했으나 당투여 1시간, 2시간 및 운동 부하 후 당투여 1시간 군등 모든 군의 당원 농도는 정상 군의 19-32%의 범위 내에 있었다. 혈당 및 인슐린은 운동 부하 후 당 투여 군에서 가장 높았으며 유리 지방산은 대조군 및 당투여 2시간군에서 높았다. 운동 부하 중 가자미근 및 적색 비복근의 당원 감소는 초 과축적 당원군, 과축적군 및 정상 군에서 다같이 운동 부하 첫 30분에 급격하였으나 초 과축적군에서 더 급격하였다. 운동 부하 30분엣 60분 사이에는 초 과축적군 및 정상 군에서 다같이 당원 감소는 거의 없었으나 과축적군에서는 지속적인 감소 상을 보였다. 간 당원은 정상 군에서 가장 급격한 감소를 보였으며 골격근과는 달리 운동부하 60분간 지속적으로 감소하였다. 초 과축적당원군의 간 당원 농도는 변화가 없거나 증가하는 경향이었다. 운동 부하 중 혈당은 모든 실험군에서 감소하였으며 인슐린은 운동 부하 후 당투여 군에서 감소하였다. 혈중 유리 지방산 농도는 모든 군에서 증가하였으며 운동 부하 후 당투여 군에서 가장 낮았다. The aim of the present investigation has been to evaluate depletion pattern of the supercompensated stores is based in the fact that a glycogen-depleted muscle by exercise with have an increased avidity for glycogen when exposed to a high carbothdrate diet. The glycongen concentration of soleus, red gastrocenemius and plantaris muscle, and liver was measured at 0,30 and 60 minutes during treadmill exercise. The experimental animals were divided into 5 group - Normal(N), Control(C), 1Hour(1HR:after 1hour of glucose ingestion), 2Hour(2HR:after 2hour of glucose ingestion )and Exercies- 1Hour(EX-1HR:glucose ingestion after 1 hour of preloading treadmill exercise)group - for glycogen storage study. The glycogen concentration of soleus, red gastrocnemius and plantaris muscles in N group was 4.57±0.34,5.11±0.24 and 6.55±0.20 mg/gm wet wt., respectively. The glycogen concentration of soleus and red gastrocnemius on EX-1HR group were about 1.9 and 1.8 times than that of N group, respectively, but the concentration of plantaris was not higher than that of N group. The glycogen concentration of liver in N group was 41.0±1.47mg/gm wet wt. and the concentration of the overnight fasted C group was only 2.9% of the value of N group. The level of glycogen concentration of liver in the other glucose ingested groups(1HR,2HR, including EX-1HR) was within 19-32% of that of N group. The blood glucose concentration of EX-1HR group was higher than that of N group, the plasma free fatty acid concentration of C and 2HR group was higher than that of N group and the plasma insulin concentration of EX-1HR group was higher than that of N group. The concentrations os supercompensated glycogen of soleus and red gastrocnemius were rapidly decreased during 30 minutes of exercise but there was almost no changes of the concentration during the other 30 minutes of continuing exercise. The concentration of N group during 30 minutes of exercise was decreased but more slowly than those of EX-1HR group. The remaining level of glycogen after 60 minutes of exercise in EX-1HR group was higher than that of N group. Taken together, the mobilization of endogenous muscle glycogen at the first stage of exercise was proportioned to the initial level of glycogen concnetration, and later on, when exercise continued, the muscle glycogen level was stabilized. And the remaining level of supercompensated muscle glycogen after 60 minutes of exercise was higher than that of normally stored glycogen level. The mobilization of the glycogen stroed in show and fast oxidative muscle fibers in faster than in the fast glycolytic muscle fibers during strenous exercise.

Glycogen Metabolism in Vibrio vulnificus Affected by malP and malQ

( Ah-reum Han ),( Yeon-ju Lee ),( Tianshi Wang ),( Jung-wan Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 한국미생물·생명공학회지 Vol.46 No.1

Vibrio vulnificus needs various responsive mechanisms to survive and transmit successfully in alternative niches of human and marine environments, and to ensure the acquisition of steady energy supply to facilitate such unique life style. The bacterium had genetic constitution very different from that of Escherichia coli regarding metabolism of glycogen, a major energy reserve. V. vulnificus accumulated more glycogen than other bacteria and at various levels according to culture medium and carbon source supplied in excess. Glycogen was accumulated to the highest level in Luria-Bertani (3.08 mg/mg protein) and heart infusion (4.30 mg/mg protein) complex media supplemented with 1% (w/v) maltodextrin at 3 h into the stationary phase. Regarding effect of carbon source, more glycogen was accumulated when maltodextrin (2.34 mg/mg protein) was added than when glucose or maltose (0.78-1.14 mg/mg protein) was added as an excessive carbon source to M9 minimal medium, suggesting that maltodextrin metabolism might affect glycogen metabolism very closely. These results were supported by the analysis using the malP (encoding a maltodextrin phosphorylase) and malQ (encoding a 4-α-glucanotransferase) mutants, which accumulated much less glycogen than wild type when either glucose or maltodextrin was supplied as an excessive carbon source, but at different levels (3.1-80.3% of wild type glycogen). Therefore, multiple pathways for glycogen metabolism were likely to function in V. vulnificus and that responding to maltodextrin might be more efficient in synthesizing glycogen. All of the glycogen samples from 3 V. vulnificus strains under various conditions showed a narrow side chain length distribution with short chains (G4-G6) as major ones. Not only the comparatively large accumulation volume but also the structure of glycogen in V. vulnificus, compared to other bacteria, may explain durability of the bacterium in external environment.

14C-glucose를 이용한 시험관 실험에서 비후 골격근의 glycogen 합성능

김용운,이석강,김종연 영남대학교의과대학 1990 Yeungnam University Journal of Medicine Vol.7 No.1

Slow twitch soleus와 fast twitch plantaris근의 운동, insulin, 전기자극에 대한 반응을 시험관내에서 ??-glucose를 이용한 glycogen합성능으로 비교하고 협력근 절제 4주후에 유도한 대상성비후근의 glycogen의 합성능을 같은 방법으로 평가하여 지속적인 과부하에 따른 각 근섬유의 반응을 비교하고자 한 본 연구의 결과를 요약하면 다음과 같다. 대조군의 soleus에서는 glycogen으로 incorporation되는 glucose양으로 볼때 운동에 의한 증가는 없었으나 insulin, 전기자극등의 인자에 의해서는 증가하였으며 운동이 이들 인자들과 합쳐 졌을때는 이들의 작용을 증폭 시켰다. 그러나 plantaris에서는 운동과 전기자극을 합한 군과 모든자극을 함께 가한 군에서만 유의한 증가를 보여서 insulin이나 glucose대사의 관점에서 볼때 soleus가 훨씬 활동적임을 알수 있었다. 협력근 절제로 인한 4주간의 과부하로 인한 비후군의 glycogen합성능은 soleus에서 대조군의 그것과 양상은 비슷하였으나 그 절대량에서 크게 못미쳤으며 plantaris에서는 대조군과 매우 유사한 반응을 나타내었다. 이와같은 결과로 볼때 협력근 절제 4주후에 여러 효소의 활동도가 안정상태에 이른다는 여러 보고에 기초하면 지속적인 과부하에 대한 각 근섬유의 반응양상이 차이가 나서 fast twitch근의 근활동이 더 향상되는 것으로 사료된다. This investigation was undertaken to clarify this in vitro effect of the various stimulations, such as exercise(E), insulin(I), direct electrical stimulation(EST) and the combinations of the above, on the glucose incorporation into glycogen molecules (glycogen synthesis)of the normal slow(soleus) and fast twitch(plantaris) muscles, and the different responses of slow and fast twitch muscles to persistent overloads causing compensatory muscle hypertrophy. In resting state, slow twitch muscle has greater capacity for glycogen synthesis than fast twitch muscle, and responses of different muscle to various stimuli were differ as follows: In slow twitch muscle, the glycogen synthesis was increased by insulin, and electrical stimulation but not increased by exercise ; exercise increased insulin sensitivity and the effect of electrical stimulation. Whereas the glycogen synthesis in fast twitch muscle was increased only by the stimuli combined with E and EST, and E,I, and EST. As the result of removal of synergistic muscle, both muscles were hypertropied, and the degree of hypertrophy in response to persistent overload was higher in fast twitch muscle(182%) than slow twitch muscle(151%). In hypertrophied muscles, glycogen synthesis of soleus in any groups was lower than that of the control, but similar in plantaris. In conclusions, there were marked heterogeneity in defferent muscle fiber in the effects of exercise and insulin addition and electrical stimulation on muscle glycogen synthesis, and fast twitch muscle may be adapted more easily to that kind of persistent overload than slow twitch muscle.

Glycogen Metabolism in Vibrio vulnificus Affected by malP and malQ

Han, Ah-Reum,Lee, Yeon-Ju,Wang, Tianshi,Kim, Jung-Wan The Korean Society for Microbiology and Biotechnol 2018 한국미생물·생명공학회지 Vol.46 No.1

Vibrio vulnificus needs various responsive mechanisms to survive and transmit successfully in alternative niches of human and marine environments, and to ensure the acquisition of steady energy supply to facilitate such unique life style. The bacterium had genetic constitution very different from that of Escherichia coli regarding metabolism of glycogen, a major energy reserve. V. vulnificus accumulated more glycogen than other bacteria and at various levels according to culture medium and carbon source supplied in excess. Glycogen was accumulated to the highest level in Luria-Bertani (3.08 mg/mg protein) and heart infusion (4.30 mg/mg protein) complex media supplemented with 1% (w/v) maltodextrin at 3 h into the stationary phase. Regarding effect of carbon source, more glycogen was accumulated when maltodextrin (2.34 mg/mg protein) was added than when glucose or maltose (0.78.1-14 mg/mg protein) was added as an excessive carbon source to M9 minimal medium, suggesting that maltodextrin metabolism might affect glycogen metabolism very closely. These results were supported by the analysis using the malP (encoding a maltodextrin phosphorylase) and malQ (encoding a 4-${\alpha}$-glucanotransferase) mutants, which accumulated much less glycogen than wild type when either glucose or maltodextrin was supplied as an excessive carbon source, but at different levels (3.1-80.3% of wild type glycogen). Therefore, multiple pathways for glycogen metabolism were likely to function in V. vulnificus and that responding to maltodextrin might be more efficient in synthesizing glycogen. All of the glycogen samples from 3 V. vulnificus strains under various conditions showed a narrow side chain length distribution with short chains (G4-G6) as major ones. Not only the comparatively large accumulation volume but also the structure of glycogen in V. vulnificus, compared to other bacteria, may explain durability of the bacterium in external environment.

Optimized M9 Minimal Salts Medium for Enhanced Growth Rate and Glycogen Accumulation of Escherichia coli DH5α

Wang, Liang,Liu, Qinghua,Du, Yangguang,Tang, Daoquan,Wise, Michael J. The Korean Society for Microbiology and Biotechnol 2018 한국미생물·생명공학회지 Vol.46 No.3

Glycogen plays important roles in bacteria. Its structure and storage capability have received more attention recently because of the potential correlations with environmental durability and pathogenicity. However, the low level of intracellular glycogen makes extraction and structure characterization difficult, inhibiting functional studies. Bacteria grown in regular media such as lysogeny broth and tryptic soy broth do no accumulate large amounts of glycogen. Comparative analyses of bacterial media reported in literature for glycogen-related studies revealed that there was no consistency in the recipes reported. Escherichia coli $DH5{\alpha}$ is a convenient model organism for gene manipulation studies with respect to glycogen. Additionally, M9 minimal salts medium is widely used to improve glycogen accumulation, although its composition varies. In this study, we optimized the M9 medium by adjusting the concentrations of itrogen source, tryptone, carbon source, and glucose, in order to achieve a balance between the growth rate and glycogen accumulation. Our result showed that $1{\times}M9$ minimal salts medium containing 0.4% tryptone and 0.8% glucose was a well-balanced nutrient source for enhancing the growth and glycogen storage in bacteria. This result will help future investigations related to bacterial physiology in terms of glycogen function.

Optimized M9 Minimal Salts Medium for Enhanced Growth Rate and Glycogen Accumulation of Escherichia coli DH5α

( Liang Wang ),( Qinghua Liu ),( Yangguang Du ),( Daoquan Tang ),( Michael J. Wise ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 한국미생물·생명공학회지 Vol.46 No.3

Glycogen plays important roles in bacteria. Its structure and storage capability have received more attention recently because of the potential correlations with environmental durability and pathogenicity. However, the low level of intracellular glycogen makes extraction and structure characterization difficult, inhibiting functional studies. Bacteria grown in regular media such as lysogeny broth and tryptic soy broth do no accumulate large amounts of glycogen. Comparative analyses of bacterial media reported in literature for glycogen-related studies revealed that there was no consistency in the recipes reported. Escherichia coli DH5α is a convenient model organism for gene manipulation studies with respect to glycogen. Additionally, M9 minimal salts medium is widely used to improve glycogen accumulation, although its composition varies. In this study, we optimized the M9 medium by adjusting the concentrations of itrogen source, tryptone, carbon source, and glucose, in order to achieve a balance between the growth rate and glycogen accumulation. Our result showed that 1 × M9 minimal salts medium containing 0.4% tryptone and 0.8% glucose was a well-balanced nutrient source for enhancing the growth and glycogen storage in bacteria. This result will help future investigations related to bacterial physiology in terms of glycogen function.

[Retracted] Epinephrine Control of Glycogen Metabolism in Glycogen-associated Protein Phosphatase PP1G/R_GLKnockout Mice

김종화,Anna A. DePaoli-Roach 생화학분자생물학회 2002 Journal of biochemistry and molecular biology Vol.35 No.3

The glycogen-associated protein phosphatase (PP1G/$R_{GL}$) may play a central role in the hormonal control of glycogen metabolism in the skeletal muscle. Here, we investigated the in vivo epinephrine effect of glycogen metabolism in the skeletal muscle of the wild-type and $R_{GL}$ knockout mice. The administration of epinephrine increased blood glucose levels from 200±20 to 325±20 mg/dl in both wild-type and knockout mice. Epinephrine decreased the glycogen synthase -/+ G6P ratio from 0.24±0.04 to 0.10±0.02 in the wild-type, and from 0.17±0.02 to 0.06±0.01 in the knockout mice. Conversely, the glycogen phosphorylase activity ratio increased from 0.21±0.04 to 0.65±0.07 and from 0.30±0.04 to 0.81±0.06 in the epinephrine trated wild-type and knockout mice respectively. The glycogen content of the knockout mice was substantially lower (27%) than that of both wild-type mice; and epinephrine decreased glycogen content in the wild-type and knockout mice. Also, in Western blot analysis there was no compensation of the other glycogen targeting components PTG/R5 and R6 in the knockout mice compared with the wild-type. Therefore, $R_{GL}$ is not required for the epinephrine stimulation of glycogen metabolism, and rather another phosphatase and/or regulatory subunit appears to be involved.

트레드밀 운동 후 과당섭취가 저장 글리코겐 회복에 미치는 영향

김장원,류승필,정혁,정현경,권태동,김형렬,이수천 한국운동영양학회 1999 Physical Activity and Nutrition (Phys Act Nutr) Vol.3 No.2

Stored glycogen is one of the important energy source during endurance exercise thus glycogen repletion, when exhausted, should be considered in most of athletes. At this point of view, using SD rats, glycogen repletion after exhausted exercise was investigated. Forty-eight male SD rats run on the treadmill until exhausted(7˚ incline, 35m/min). When exhausted, they were given ad-libitum either fructose (5%/100㎖, experimental group; Exe) or water (control group: Con) as a placebo for 60min or 120min. Rats were decapitated at pre-exercise, exhausted, 60min, and 120min of recovery, respectively. Serum glucose, lactate, insulin, FFA, muscle glycogen and liver glycogen was analyzed. Glucose concentration in Exe was significantly higher than Con (p<.05) during recovery periods. Lactate and insulin concentrations were not affected to both experimental substrates. FFA concentration was decreased at 120min of recovery in Exe but was continuously increased in Con. Muscle glycogen concentration was significantly repleted by fructose ingestion compared to water ingestion. Liver glycogen was, also, significantly repleted in Exe and its ratio of repletion was faster than muscle glycogen. These results suggested that, like other carbohydrate solutions such as glucose, sucrose, and maltodextrine and so forth, fructose ingestion after exhausted exercise has beneficial effect for depleted glycogen repletion compared to water ingestion.

Characterization of the Transglycosylation Reaction of 4-α-Glucanotransferase (MalQ) and Its Role in Glycogen Breakdown in Escherichia coli

( Dang Hai Dang Nguyen ),( Sung-hoon Park ),( Phuong Lan Tran ),( Jung-wan Kim ),( Quang Tri Le ),( Winfried Boos ),( Jong-tae Park ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.3

We first confirmed the involvement of MalQ (4-α-glucanotransferase) in Escherichia coli glycogen breakdown by both in vitro and in vivo assays. In vivo tests of the knock-out mutant, ΔmalQ, showed that glycogen slowly decreased after the stationary phase compared to the wild-type strain, indicating the involvement of MalQ in glycogen degradation. In vitro assays incubated glycogen-mimic substrate, branched cyclodextrin (maltotetraosyl-β-CD: G4-β-CD) and glycogen phosphorylase (GlgP)-limit dextrin with a set of variable combinations of E. coli enzymes, including GlgX (debranching enzyme), MalP (maltodextrin phosphorylase), GlgP and MalQ. In the absence of GlgP, the reaction of MalP, GlgX and MalQ on substrates produced glucose-1-P (glc-1-P) 3-fold faster than without MalQ. The results revealed that MalQ led to disproportionate G4 released from GlgP-limit dextrin to another acceptor, G4, which is phosphorylated by MalP. In contrast, in the absence of MalP, the reaction of GlgX, GlgP and MalQ resulted in a 1.6-fold increased production of glc-1-P than without MalQ. The result indicated that the G4-branch chains of GlgP-limit dextrin are released by GlgX hydrolysis, and then MalQ transfers the resultant G4 either to another branch chain or another G4 that can immediately be phosphorylated into glc-1-P by GlgP. Thus, we propose a model of two possible MalQ-involved pathways in glycogen degradation. The operon structure of MalP-defecting enterobacteria strongly supports the involvement of MalQ and GlgP as alternative pathways in glycogen degradation.