Developmental expression of glycogenolytic enzymes in rabbit tissues: possible relationship to fetal lung maturation.
Glycogen can be degraded in mammalian tissues by one of three isozymes of glycogen phosphorylase, termed muscle (M), liver (L) and brain (B) after the tissues in which they are preferentially expressed in adult animals, or by members of the family of alpha-glucosidases. In the current study, we have examined the developmental expression of these enzymes and their respective mRNAs in rabbit tissues, with particular emphasis on the developing lung, a tissue in which glycogen serves as an important source of carbon for surfactant phospholipid biosynthesis. Native gel activity assays and RNA blot hybridization analysis revealed that the B isoform of glycogen phosphorylase predominates in fetal and adult lung tissues, accompanied by a low level of expression of the M isoform. Total B and M phosphorylase activities increased during fetal lung development, with a peak at day 28 of gestation, then decreased to the adult level at term. This peak in activity coincided with the peak period of glycogen degradation in developing lung. While the increase in M isozyme activity was correlated with an increase in the level of its mRNA, B isoform mRNA showed no significant alteration during development, suggesting that the increase in B isoform activity is determined by a posttranscriptional mechanism. Analysis of phosphorylase mRNA levels in developing liver, skeletal muscle, brain and heart revealed a diverse expression pattern. The L isozyme mRNA was predominant at all time points in liver, the M isozyme was predominant at all time points in muscle, the B isozyme was predominant at all time points in brain, and heart contained a mixture of B and M mRNA in roughly equal ratios at all time points. Thus, our studies of phosphorylase mRNA in the rabbit provide no evidence for general predominance of the B isozyme in fetal tissues, or for isozyme 'switching' from the B to the L or M forms during development, as has been suggested by others. In addition to the increase in phosphorylase activity, acid, but not neutral alpha-glucosidase activity was found to increase significantly during fetal lung development, again with a peak at day 28 of gestation. Interestingly, RNA blot hybridization analysis with a probe for lysosomal alpha-glucosidase revealed no change in the level of expression of its 4 kb transcript in developing lung. Instead, we observed induction of a structurally related mRNA of 7.4 kb that peaked at day 28 of gestation. Hybridization with a sucrase/isomaltase-specific oligonucleotide excluded the possibility that the 7.4 kb transcript encodes this protein.(ABSTRACT TRUNCATED AT 400 WORDS)
Newgard
C B
CB
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235.
Norkiewicz
B
B
Hughes
S D
SD
Frenkel
R A
RA
Coats
W S
WS
Martiniuk
F
F
Johnston
J M
JM
D000818:Animals; D001483:Base Sequence; D004247:DNA; D005314:Embryonic and Fetal Development / Q000235:genetics*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006003:Glycogen / Q000378:metabolism; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology* / Q000254:growth & development; D051379:Mice; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity / Q000235:genetics; D006005:Phosphorylases / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits
D006003:Glycogen; D004247:DNA; D006005:Phosphorylases