Dynamics of non-specific esterase during fat resorption in the jejunum of the house mouse,Mus musculus

Summary The dynamics of non-specific esterase in the upper duodenum of the house mouse was studied electron microscopically at various intervals following a fat meal. Enterocytic esterase became associated with lipid droplets during fat resorption and formation of primary chylomicrons. Esterase activity remained associated with the primary chylomicrons throughout the process of extrusion into the extracellular space at the lateral interdigitations, and during subsequent transport into the lymph vessels. It is suggested that certain isozymes of non-specific esterase participate in lipid transport.Supported by the Deutsche Forschungsgemeinschaft (SFB 46). This is contribution no. 37 of a research program devoted to the cellular distribution and genetics of non-specific esterase     

Electron microscopical demonstration of sulphated mucopolysaccharides in mouse tracheal cartilage with a diaminobenzidine-osmium tetroxide technique

Synopsis A diaminobenzidine-osmium tetroxide method for the demonstration of sulphated mucopolysaccharides has been tested at the electron microscopical level. The reaction (which involves treatment with diaminobenzidine in an acidic solution followed by oxidation with osmium tetroxide) has been carried out directly on ultra-thin sections of mouse tracheal cartilage embedded in water-soluble glycolmethacrylate. Sulphated mucopolysaccharides in the cartilage matrix were localized as a heavy, electron-dense precipitate. The method can be applied directly to sections, overcoming in this way the difficulties of penetration, and it seems to possess a higher specificity for sulphated mucopolysaccharides than that displayed by other techniques proposed previously for the ultrastructural demonstration of acid mucopolysaccharides.     

Lymph esterases of the house mouse (Mus musculus)--I. Identification and possible origins of abdominal lymph esterases

1. Abdominal lymph was obtained from Mus musculus by cannulation of the thoracic duct: lymph esterases were identified by polyacrylamide gel electrophoresis. Seven known esterases (ES-1, ES-2, ES-5, ES-27, SE-I, SE-II and SE-III) and a newly described activity (SE-IV) were demonstrated, all of which were also present in serum. 2. Electrophoretic staining intensities indicated that the lymph esterases were less concentrated than the corresponding activities in serum, with the single exception of ES-2. This finding was supported by quantitative immunoelectrophoresis of ES-1 and ES-2 (two allozymes each). 3. The jejunum appeared to be the origin of lymph ES-2 by a comparison of organ distribution of the allozymes ES-2B and ES-2D and by monitoring the re-appearance of ES-2 in several organs, serum and lymph after total inhibition in vivo by bis-p-nitrophenyl phosphate.     

Electron microscopical demonstration of thiols and disulphides in the porcine epidermis

Summary This study describes the electron microscopical distribution of free thiols and disulphides in the epidermis of the domestic pig and the wild boar, as compared to light microscopical histochemistry. With the silver methenamine method, silver labelling of thiols was clearly achieved on the keratohyalin and cytofilament accumulations in the cells of the living epidermis and the plasma membrane of granular cells. To a certain extent, the envelope and cytoplasm of young corneocytes reacted equally intensively. Disulphides were very abundant in the filaments, keratohyalin granules, and cell envelope of granular cells, and, particularly, in the envelope (marginal band) of corneal cells; the latter structure being distinctly delineated from the background. As a specific feature, the viable epidermis of the wild boar stained strongly for disulphides. The results obtained are discussed in view of actual concepts of epidermal keratinization and corneal cell function.     

Esterase XX. Disc-electrophoretic investigations on the polymorphism of the esterases of the house mouse (author's transl)]

For the further clarification of the polymorphism of mouse-esterase and its hormonal control, which in part have not yet been fully comprehended, disc-electrophoretic analyses of eight organs were made, using a strain with the Tfm-mutation. In addition, quantitative assays of esterase activity as well as histochemical studies were performed. The individual organs are characterized by a specific banding pattern of esterase, which is essentially conditioned by the diverse activity of a limited number of bands. Partly these may be regarded as primary gene products, partly they seem to be secondary modifications. The few incidences of band-linkage justify the expectations, that further gene loci will be discovered. In four organs of Tfm-mutants a lower esterase activity was found than in the controls, which was especially distinct in the kidney. The behaviour of the testosterone-dependent bands in the kidneys of Tfm-mutants seems to indicate two different mechanisms of the effect of testosterone on these bands.     

Electron microscopical demonstration of acid phosphatase activity in the central nervous system

Summary A modified technique is described for the demonstration of acid phosphatase activity in the central nervous system by means of electron microscopy. Enzyme activity can be demonstrated in lysosomes, pigment bodies, and the Golgi zone of cortical neurons. Glial and endothelial cells also contain acid phosphatase active lysosomes. They are located in the pericarya, and in the processes of the glial cells, respectively.The authors express their sincere appreciation to FräuleinS. Luh and FräuleinW. Komp for their assistance and help.     

Lymph esterases of the house mouse (Mus musculus)--II. The role of esterase-2 in fat resorption

1. Intralipid infusion into the duodenum of Mus musculus was accompanied by changes in lymph and serum concentrations of two esterase isozymes, ES-1 and ES-2. Whereas ES-1 levels declined in both lymph and serum, ES-2 levels increased 5-fold in lymph within 120 min, and fell to a plateau 3- to 4-fold the fasting level; serum levels of ES-2 increased continually. 2. The changes in lymph ES-2 concentrations were paralleled by lymph triglyceride concentration during Intralipid infusion. Genetically determined differences in the concentration of two allozymes, ES-2B and ES-2D, were reflected in differences in lymph triglyceride levels. The lymph triglyceride concentration was strongly correlated with approximately the cube root of the lymph ES-2 concentration for both allozymes. 3. The source of lymph ES-2 during fat resorption was probably an intracellular jejunal pool; serum ES-2 also re-entered the lymph but this fraction was not influenced by fat resorption. 4. Purified chylomicrons possessed no esterase activity; however, it was postulated that ES-2 plays an essential role in fat resorption and is extruded with the primary chylomicrons from the enterocyte.     

Electron microscopical demonstration of polysaccharides in rotifers (Aschelminthes) with the use of OsO4-ferricyanide (author's transl)]

Three species of rotifers were fixed with glutaraldehyde and postfixed in a OsO4-ferricyanide solution. This procedure results in staining of the sarcoplasmic reticulum (SR) and the surface coat of muscle cells as well as in staining of the surface coat of glands and nerve cells. Section-staining with leadcitrate increased the electron opacity of the precipitates, which are interpreted as polysaccharides. The partial decrease of staining of polysaccharides, which sometimes is observed, may be related to processes of function. The precipitates within the SR are considered as glycogen-beta-particles.     

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.     

Electron microscopical demonstration of glucose-6-phosphatase in native cryostat sections fixed with glutaraldehyde through semipermeable membranes.

The cytochemical demonstration of glucose-6-phosphatase (G6Pase) activity in native cryostat sections fixed with glutaraldehyde through semipermeable membranes is superior to conventional methods with regard to exact localization and lack of inactivation and diffusion of the enzyme, together with simultaneous excellent preservation of the tissue fine structure. In rat liver not only hepatocytes but also many bile duct epithelia and endothelia of arterioles and venules show a marked G6Pase activity in the membranes of the endoplasmic reticulum including the nuclear envelope.     

In vitro studies on the effects of several insecticides on non-specific esterases of Sitophilus oryzae (L.) and Palembus dermestoides (fairm.)

1. Sitophilus oryzae (Curculionidae) and Palembus dermetoides (Tenebrionidae) esterases were studied to determine the correlation between in vitro esterase inhibition and in vivo toxicity of several insecticides.2. K_m values for S. oryzae and P. dermetoides were 0.118 mM and 0.087 mM respectively.3. In vitro esterase inhibition by organophosphates and carbamates showed that for S. oryzae dichlorvos (I₅₀ = 7 × 10⁻⁶ M) and eserine sulphate (I₅₀ = 5 × 10⁻⁵ M) were most potent; for P. dermestoides it was methomyl (I₅₀ = 2.8 × 10⁻⁷ M) and dichlorvos (I₅₀ = 10⁻⁶ M). For both species chlorpyrifos-methyl was least so.4. Only slight correlation between in vitro esterase inhibition and in vivo toxicity of the insecticides could be shown for either species. This is discussed.     

Electron microscopical demonstration of glucose-6-phosphatase in native cryostat sections fixed with glutaraldehyde through semipermeable membranes

Summary The cytochemical demonstration of glucose-6-phosphatase (G6Pase) activity in native cryostat sections fixed with glutaraldehyde through semipermeable membranes is superior to conventional methods with regard to exact localization and lack of inactivation and diffusion of the enzyme, together with simultaneous excellent preservation of the tissue fine structure. In rat liver not only hepatocytes but also many bile duct epithelia and endothelia of arterioles and venules show a marked G6Pase activity in the membranes of the endoplasmic reticulum including the nuclear envelope. This work was kindly supported by the Deutsche Forschungsgemeinschaft An erratum to this article is available at .     

Some characteristics of the XO mouse (Mus musculus L.) I. Vitality: Growth and metabolism

The growth rate of XO mice during the first five weeks of life was shown to be significantly lower (ca. 15%) than the growth rate of normal XX mice. A marker gene Tabby was introduced in order to recognize hemizygous XO females. The presence or absence of this gene had a significant influence on growth rates. XO females could only be compared to XX females in an indirect way. The differences found could not be attributed to maternal influence or to the influence of litter size.Body temperature and thyroid activity were found to be lower in XO mice than in normal females. It is suggested that the lower growth rate characteristic of the XO mice is a consequence of hypothyroidism and a lower basal metabolic rate.The results show that phenotypically XO mice are not entirely normal and at least two normal X's are necessary for complete development.     

On the fungitoxicity of some new thiocyanatopyrazole derivatives: Electron microscopical study in trichophyton mentagrophytes

Four thiocyanatopyrazole derivatives were synthesized and their fungistatic activity was demonstrated in vitro against a number of dermatophytic fungi. In Trichophyton mentagrophytes, the most active compound induced an unusual increase of the plasma membrane with production of intra and extracytoplasmic complexes, a deterioration of nuclear and mitochondrial membranes and a formation of autophagic-like vacuoles. Plasmolysis, accompanied by an almost complete disorganization of cytoplasmic structures, seemed to be the final event. A possible mechanism of action of the compounds was discussed.Investigation supported by a grant from Consiglio Nazionale delle Ricerche of Italy (Contract No. 7500536).     

Genetic control of two electrophoretic variants of glucosephosphate isomerase in the mouse (Mus musculus)

The autosomal variation and the genetic control of GPI has been determined by a comparison of electrophoretic patterns of F₁ and backcross progeny of three inbred strains of mice. The locus controlling the production of GPI in the mouse has been designated Gpi-1. Two alleles at this locus have been described and designated Gpi-1 ^a and Gpi-1 ^b, which represent, respectively, the slow and fast electrophoretic forms. Twenty-seven inbred strains of mice have been classified for these two alleles. The absence of close linkage of Gpi-1 to seven other genetic loci has been determined. It has been demonstrated that the polymorphism of Gpi-1 is widely distributed in feral mice. GPI was expressed in vitro and in four types of malignant tumors.Supported by U.S. Public Health Service Grants GM-09966, from General Medical Sciences, and GY 4193.     

Some characteristics of the XO mouse (Mus musculus L.) II. Reproduction: fertility and gametic segregation

A preliminary investigation of reproductive capacity in XO mice showed that they produced smaller litters than normal litter mates. More time elapsed between successive litters when kept in the presence of a male except during pregnancy and weaning. This lower level of reproduction is manifest during the whole reproductive period. Also reproductive capacity in XO mice reaches its maximum and minimum (end of reproductive period) earlier than in normals.Further study also demonstrated an underdevelopment of the ovaries-the whole ovary, as well as the numbers of maturing and mature follicles, are smaller in XO mice than in controls. Since these differences can be corrected by unilateral ovariectomy, they appear to be under control of extra-ovarial factors, e.g. of gonadotropic hormone(s). The data also suggest depression of activity in the oestrogen-producing system, and in general that the lower reproductivity of XO mice may be attributed to a diminished secretion of gonadotropic hormones together with a smaller number of primordial germ cells in XO mice.The segregation from XO parents seems to favor transmission of X-gametes in young mothers, with however the preference for X tending to decrease with parental age. Since the data tend to rule out postzygotic selection effects, the excess of X-bearing gametes may relate to meiotic drive.     

Polymeric hemoglobins of the house mouse (Mus musculus L.): Isolation of cysteinyl peptides

Hemoglobins from wild populations of the house mouse (Mus musculus L.) are polymorphic with respect to the diffuse or single appearance of their electrophoretic patterns and their ability to form polymers. Polymerization occurs by the formation of intermolecular disulfide bonds. Isolation of all the cysteinyl peptides shows that the reactive cysteinyl residue, 13, is in the same position as that found in BALB/cJ laboratory mice.This research was supported in part by grant F-213 from the Robert A. Welch Foundation, and NIH grants GM-05818 and GM-09326, and an NIH predoctoral fellowship to J. B.