Esterase-16 (Es-16): Characterization,polymorphism, and linkage to chromosome 3 of a kidney esterase locus of the house mouse

A polymorphism for an isozyme of a presumed arylesterase, esterase-16 (EC 3.1.1.2), has been detected in kidney, heart, and spleen of the house mouse, Mus musculus, by means of isoelectric focusing and by disc electrophoresis. Three phenotypes can be distinguished: the ES-16A phenotype (IEP 5.9) was found in C57BL/10Sn and many other laboratory inbred strains; the ES-16B phenotype (IEP 6.1) was found in M. m. molossinus; and the ES-16C phenotype (IEP 5.9; very weak activity) was found in Peru-Coppock. Esterase-16 is strongly inhibited by 10^?3 m p-chloromercuribenzoate, but not by 2·10^?4 m bis-p-nitrophenyl phosphate or by 10^?3 m Diamox. It stains well with indoxyl acetate and other indigogenic substrates but only weakly with α-naphthyl acetate. Esterase-16 is completely insoluble in water. It is apparently governed by a structural gene locus, Es-16, with three alleles, Es-16 ^a , Es-16^b, and Es-16 ^c, respectively. Es-16 is closely linked to Car-1 and Car-2 on chromosome 3. Typing of 94 animals of the backcross (C57BL/10Sn × M. m. mol.) F₁ × M. m. mol. revealed a recombination frequency of 8.51±2.9%.     

Polymorphism of esterase 11 in Mus musculus,a further esterase locus on chromosome 8

A further esterase, esterase 11, which exhibits a polymorphism detectable by electrophoresis, has been observed in the house mouse, Mus musculus. In 15 inbred strains and two outbred strains, the ES-11A phenotype has been found, composed of two bands of enzyme activity of greater anodal electrophoretic mobility than the two bands of the ES-11B phenotype found in one inbred strain, one wild stock, and 101 wild mice. In F₁ hybrids (IS/Cam×C57 BL/Gr), the phenotype shown corresponds to a mixture of the two parental phenotypes. In backcrosses, ES-11 segregates as an autosomal gene, designated Es-11, closely linked to Es-2 and Es-5 on chromosome 8.This work was supported by the Medical Research Council.     

Esterase-26 (ES-26): Characterization and genetic location on chromosome 3 of an eserine-sensitive esterase of the house mouse (Mus musculus)

Genetic variation of a codominantly inherited esterase, designated ES-26, has been discovered in the house mouse using isoelectric focusing in polyacrylamide gels. The ES-26A phenotype (pI 8.2) was found in C57BL/10Sn. A/J showed the ES-26B phenotype (pI 7.8-7.9). A third phenotype, ES-26C (double-banded: pI's 8.1 and 8.3), was observed in SJL/J. ES-26 was detected only in liver, stomach, and small intestine. The enzyme was shown to be controlled by the presumed structural locus Es-26, located on chromosome 3. From a four-point cross, the gene order Car-2--6.2 +/- 2.7--Es-16--21.0 +/- 4.5--Es-26--13.6 +/- 3.8--Amy-1 was established.     

Esterase-17 (ES-17): Characterization and genetic location on chromosome 9 of a bis-p-nitrophenyl phosphate-resistant esterase of the house mouse (Mus musculus)

Genetic variation of a new codominantly inherited esterase, designated ES-17, has been discovered in the house mouse using isoelectric focusing in polyacrylamide gels. The ES-17 A phenotype (three bands; isoelectric points, betweenpH 5.55 andpH 5.90) was found in C57 BL/10Sn. LP/J possessed the Es-17B phenotype (three bands; isoelectric points,pH 5.05–5.55). ES-17 was present in all tissues examined, except for hemolysate and serum, and was most clearly expressed in the small intestine. Because of its reaction toward various substrates and inhibitors, ES-17 has tentatively been classified as acetyl esterase (EC 3.1.1.6). ES-17 was shown to be controlled by the structural locusEs-17, located on chromosome 9. From test-cross data, a gene order ofEs-17-8.7±2.5 map units-Mpi-1-10.2±2.7 map units-Mod-1 was established. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 46). This is communication No. 35 of a research program devoted to the cellular distribution and genetics of nonspecific esterases.     

Esterase-6 (Es-6) in laboratory mice: Hormone-influenced expression and linkage relationship to oligosyndactylism (Os), esterase-1 (Es-1), and esterase-2 (Es-2) in chromosome 8

Allelic differences at an esterase locus designated Es-6 exist between mouse strain C57 BL/6J and a laboratory stock of M.m. molossinus. Strain C57BL/6J has been assigned the allele Es-6a and M. m. molossinus the alternate allele Es-6b. Kidney expression of the electrophoretic esterase band controlled by the Es-6 locus is sex influenced, with increased activity apparently induced by testosterone. A four-point test cross established the gene order Os-Es-1-Es-6-Es-2 within a 10-cM segment on chromosome 8.     

Genetic study of pancreatic proteinase in mice (Mus musculus): Linkage of the Prt-2 locus on chromosome 8

The Prt-2 locus is linked with Es-1 and Es-2 loci on chromosome 8 (linkage group XVIII). Recombination frequencies were 8.2% between Es-1 and Es-2, 12.7% between Es-1 and Prt-2, and 4.5% between Es-2 and Prt-2. From these data, the map position of Prt-2 has been estimated on chromosome 8. The Prt-1 and Prt-3 loci, which are linked very closely on the same chromosome, were not determined.     

Esterase-18 (ES-18) of the house mouse (Mus musculus): Biochemical characterization and genetics of an allozyme system linked to chromosome 19

This study describes the biochemical characterization, genetic variation, and linkage of a codominantly inherited murine esterase, termed ES-18. The enzyme was identified by isoelectric focusing of supernatants obtained after centrifugation of tissue homogenates and subsequent staining for esterase using either alpha-naphthyl acetate or 4-methylumbelliferyl elaidate as substrate. ES-18 exhibited an organ-specific variation of the intensity pattern of bands as seen in kidney, spleen, and macrophages, respectively. Its activity was highly sensitive to inhibition by 1 mmol.liter-1 p-chloromercuriphenylsulfonate but was resistant to bis-p-nitrophenyl phosphate. Four allozymes could be distinguished in kidney supernatants obtained from the inbred strains C57BL/10Sn (ES-18A), MOLF/Ei (ES-18B), WLL/BrA (ES-18C), and CAST/Ei (ES-18D). The enzyme is shown to be controlled by a structural locus, Es-18, which resides on chromosome 19. The gene order Ly-1 - Got-1 - 4.7 +/- 1.6 - Es-18 is suggested.     

Biochemical genetics of Es-14 (formerly Es-Si) and a new esterase variation,Es-15, of the laboratory rat (Rattus norvegicus): Biochemistry,tissue expression,and linkage to Es-1 in linkage group V

The segregation of rat esterases controlled by loci residing in linkage group V (LGV) has been studied in two backcross series, (LEW/Han × BN/Han)F₁ × LEW/Han and (LEW/Han × LE/Han)F₁ × LEW/Han. Es-14 (formerly Es-Si) was shown to be closely linked to Es-1. A new esterase locus, Es-15, was described which codes for a liver isozyme. The distribution pattern of three alleles at the Es-15 locus is presented for 52 independent inbred strains. Close linkage of Es-15 to Es-14 and to Es-1 was established, proposing the following gene order: [Es-2, Es-10]—[ES-1, ES-14, ES-15]. The esterase loci on LGV are thus separated into two gene clusters, cluster 1 and cluster 2. These conclusions are supported by the strain distribution patterns of the two RI strain series, LXB and DXE.Otto von Deimling was supported by the Deutsche Forschungsgemeinschaft (De 315/2-1, communication No. 56).     

Purification and characterization of esterase 1F, the albumin esterase of the house mouse (Mus musculus)

Esterase 1F was isolated from mouse serum and purified by ion-exchange chromatography, isoelectrofocusing, and molecular sieve chromatography. It is considered to be a glycoprotein with an apparent molecular weight of 75 000. The equivalent weight (approximately equal to 77 000 X g/mol) was estimated by titration of the catalytic site with diethyl p-nitrophenyl phosphate. The Michaelis constant Km and the catalytic constant kcat of the enzyme for 4-nitrophenyl hexanoate were determined. Esterase 1F is characterized by its ability to split a wide spectrum of substrates and its relatively low turnover rates towards the substrates tested. It belongs to the isozyme system of carboxylesterase (EC 3.1.1.1) coded for by chromosome 8. Esterase 1F was compared with three other genetically related isozymes, esterase 2, esterase 7 and esterase 9, with respect to some physical and catalytic properties.     

Mapping of nucleoside phosphorylase (Np-1) and esterase 10 (Es-10) on mouse chromosome 14

A method for detecting two alleles at Np-1 (nucleoside phosphorylase) and three alleles at Es-10 (esterase 10) from mouse blood by cellulose acetate electrophoresis is described. The allelic constitution at these loci for 44 inbred strains and stocks was determined. The location of Np-1 on chromosome 14 was established by backcross experiments in which alleles at Np-1 and Robertsonian translocations were segregating. Es-10 was shown to be linked to Np-1, and the following genetic map of Chr 14 was constructed: centromere-(8.9±4.0 cM)-[Np-1, Wc]-(10.2±1.9 cM)-Es-10-(15.5±3.7 cM)-s. The homologous human loci, NP and ES-D, are not linked.This work was supported by Contract E(11-1)-3267 with the Energy Research and Development Administration, by Contracts NO1-ES4-2156 and NO1-ES4-2159 with the National Institute of Environmental Health Sciences, and by Grants GM 19656 and GM 20919 from the National Institute of General Medical Sciences. D. A. K. was a participant in the 1975 Summer Program for College, Graduate, and Medical Students, which was supported, in part, by the Clark Foundation. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

Kidney esterases of Mus musculus: Further polymorphism of esterase-6, esterase-9, and a new esterase,esterase-20

The comparison of results obtained by different separation and staining techniques permits the definition of esterase-6 in comparison with esterase-9 and a new esterase, esterase-20. Alleles of Es-6 affect the product's ability to aggregate. Esterase-20 may be an aggregated product of Es-9. The close linkage of Es-6 and Es-9 is confirmed. Homology of esterase-6 with esterases from other mammalian species is also suggested.HRN was supported by the Medical Research Council. This is communication No. 32 of a research program devoted to the cellular distribution, genetics, and regulation of nonspecific esterases.     

Esterase-13 of the rat (Rattus norvegicus) and its homology with esterase-3 of the house mouse (Mus musculus)

A new allele of esterase-13 was detected in various laboratory inbred strains of Rattus norvegicus and designated Es-13c. The activity of ES-13 towards a range of chromogenic substrates, inhibitor profile, isoelectric points and retardation coefficients on polyacrylamide gel electrophoresis were determined. The organ specific expression of ES-13 alleles was investigated and it was shown that kidney homogenates contained a factor which modified the liver enzyme banding pattern in vitro. The features of ES-13 from the rat indicated homology between this esterase and ES-3 from the house mouse, Mus musculus domesticus.     

Cytochemical localization of non-specific esterase and acid phosphatase in spermatozoa of the mouse (Mus musculus)

Summary This paper describes methods for the cytochemical demonstration of non-specific esterase and acid phosphatase activities in spermatozoa of the mouse. The acetate and phosphate esters of 1-naphthol were used as substrates, and hexazonium pararosanilin was used as coupler in both techniques. Specificity was assured by the use of appropriate positive and negative controls. Best results were obtained with unfixed smears which had been stored at room temperature for a few days prior to use. Chemical fixation, especially with formol-calcium solutions, is not recommended for use with rodent spermatozoa. According to our investigations the murine acrosome contains very low levels of non-specific esterase and acid phosphatase which are not amenable to detection by the standard methods employing short incubations and/or with material fixed in a formalin-containing fixative.These studies have been supported by funds from the office of General Research, The University of Georgia.The authors thank Dr. J. Travis for gifts of trypsin inhibitors, Dr. H. A. Kent for the provision of Syrian hamsters, and Dr. W. J. Humphreys, Director, Electron Microscopy Laboratory, The University of Georgia, for use of photographic facilities. The senior author is indebted to Drs. Ellen M. Rasch and L. Ornstein for helpful discussions and advice concerning the development of the techniques described in this paper.Dr. Unnithan was the recipient of a postdoctoral award from the University of Georgia.     

Esterase-25(Es-25): Identification and characterization of a new kidney arylesterase of the house mouse,genetically linked toLy-18 on chromosome 12

Genetic variation of a codominantly inherited kidney esterase, designated ES-25, has been discovered in the house mouse using disc electrophoresis. The ES-25A phenotype was found in A strains, AKR, and BALB/c. ES-25B was found in C57BL strains and several other laboratory strains. The enzyme was shown to be controlled by a presumed structural locus, Es-25. The high concordance in 48 RI strains of Es-25 with Ly-18 indicated the location of Es-25 on chromosome 12. The gene order Es-25-Ly-18-D12Nyul-Pre-1 was proposed.     

Purification and characterization of esterase 2B of the house mouse, Mus musculus

The N-terminal tryptic peptide of Crithidia oncopelti cytochrome c557 X-Pro-Me3Lys-Ala-Arg in which X represents an unknown N-terminal blocking group was characterized by electrophoresis at pH 2 and by 1H and 13C nuclear magnetic resonance. 1H-NMR spectra of the tryptic peptide suggested that the blocking group X was N,N-dimethylproline although the electrophoretic mobility of the peptide suggested a larger molecular weight. The peptides X-Pro-Me3Lys and X-Pro were generated by treatment of the tryptic peptide with thermolysin and carboxypeptidase and the free blocking group X was prepared by acid hydrolysis. Comparison of the 1H-NMR spectra of these peptides with spectra of synthetic N,N-dimethylproline and N,N-dimethylprolylproline demonstrated that the blocking group was indeed N,N-dimethylproline. The 13C-NMR spectrum of the tryptic peptide was consistent with this conclusion although unambiguous assignments to all resonances could not be obtained because of the small amount of material available. The origin of the dimethylproline blocking group is discussed.     

Double insertion of homogeneously staining regions in chromosome 1 of wild Mus musculus musculus: effects on chromosome pairing and recombination

An examination of the meiotic pattern of chromosome 1 isolated from a feral mouse population and containing a double insertion (Is) of homogeneously staining regions (HSRs) was carried out. The region delineated by the proximal breakpoint of Is(HSR;1C5) 1Icg and the distal breakpoint of Is(HSR;1E3)2Icg is desynapsed during the early pachytene stage and heterosynapsed at the midpachytene, as shown by electron microscopic analysis of synaptonemal complexes. The HSRs have no effect on the segregation of chromosome 1 in heterozygous mice. The lack of homosynapsis in the region under study causes chiasmata redistribution in heteromorphic bivalents. In normal males, single chiasmata are located in the medial part of the chromosome. In heterozygotes, this segment is heterosynapsed and unavailable for recombination. This leads to a significant decrease in the frequency of bivalents bearing single chiasmata. The total number of chiasmata per bivalent is much higher in heterozygous males than in normal ones. The recombination frequency between proximal markers fz and In also is higher in heterozygous animals. The increase in the total chiasma number in the heteromorphic bivalent is due to the addition of double chiasmata located mostly at precentromeric and pretelomeric regions of the chromosome.     

A microsatellite polymorphism associated with the PLC1 (phospholipase C) locus: identification, mapping, and linkage to the MODY locus on chromosome 20.

A highly polymorphic (dC-dA)n.(dG-dT)n dinucleotide repeat at the PLC1 locus on human chromosome 20 has been identified. Primers flanking the dinucleotide repeat were used for PCR amplification of the repeat region in 37 informative kindreds from the Centre d'Etude du Polymorphisme Humain. Two-point linkage analysis indicates that PLC1 is closely linked to several chromosome 20 markers, including D20S16 (Zmax = 41.25; theta = 0.07), D20S17 (Zmax = 42.81; theta = 0.09), and ADA (Zmax = 57.24; theta = 0.05). Multipoint linkage analysis places the PLC1 locus between D20S18 and D20S17, 11.2 and 6.6 cM, respectively, from these loci (sex-averaged distances). In addition, the PLC1 gene shows linkage to the maturity-onset diabetes of the young (MODY) locus on chromosome 20 with a lod score of 4.57 at theta = 0.089.     

Expression of glucose phosphate isomerase in interspecific hybrid (Mus musculus × Mus caroli)

Hybrid Mus musculus × Mus caroli embryos were produced by inseminating M. musculus (C57BL/Ola Ws) females with M. caroli sperm. Control M. caroli embryos developed more rapidly than did control M. musculus embryos and implanted approximately 1 day earlier. At 1 1/2 days, both the hybrid embryos and those of the maternal species (M. musculus) had cleaved to the 2-cell stage. By 2 1/2 days some of the hybrids were retarded compared to M. musculus, and by 3½ days most were lagging behind. This is consistent with the idea that the rate of development of hybrid embryos declines once it becomes dependent on embryo-coded gene products. We have used this difference in rate of preim-plantation development, between hybrid and M. musculus embryos, to try to determine whether the activation of embryonic Gpi-1s genes, that encode glucose phosphate isomerase (GPI-1), is age-related or stage-related. In control M. musculus embryos (both mated and Al groups), the GPI-1AB and GPI-1A allozyme, indicative of paternal gene expression, were detected in 7 of 9 samples of 3 1/2-day compacted morula stage embryos and were seen in all 19 samples of 31/2-day blastocysts. In hybrid embryos, these allozymes were detected 1 day later. They were not detected in any 31/2-day samples (12 samples of compacted morulae) but were consistently detected at 4½ days (4 samples of blastocysts and 2 samples of uncompacted morulae). Our interpretation of the results is that gene activation in hybrid embryos is stage-specific, rather than age-specific, and probably begins around the 8-cell stage, with detectable levels of enzyme accumulating later. Analysis of GPI-1 elec-trophoresis indicated that both the paternal (M. caroli) and maternal (M. musculus) Gpi-1s alleles were equally expressed in hybrid embryos and that the paternally derived allele was not activated before the maternally derived allele. © 1992 Wiley-Liss, Inc.     

Serum esterase genetics in rats: Two new alleles at Es-2, a new esterase regulated by hormonal factors,and linkage of these loci to the Ag-C blood group locus

Two new alleles at the Es-2 locus are described which determine electrophoretic variants of serum esterases of rats. A new esterase protein is described which is detectable in sera of sexually mature females of the appropriate genotype. Evidence is presented for genetic linkage between the Ag-C blood group locus and Es-1, Es-2, and the locus controlling the sex-influenced protein. Since the Ea-1 blood group locus of mice is linked to four esterase loci, it is suggested that Ag-C is the rat homologue of the mouse Ea-1 locus.This work was supported by U.S.P.H.S. Grants AI-09275, CA-15146, and CA-10097.