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Biomedical subjects

P Mak

Publications and source records attributed to P Mak.

42 records · Page 3Linked to original sources

Characterization of an estrogen receptor in the turtle testis.

Using DNA-cellulose affinity chromatography, an estrogen-binding component having the properties of a classical estrogen receptor was characterized from testicular cytosol of the freshwater turtle, Chrysemys picta. This putative cytoplasmic receptor exhibited high affinity (Kd = 7.0 X 10(-10) M), low capacity (1-4 fmol/mg protein), and steroid binding that was specific for estrogens. It was not present in plasma, muscle, kidney, or lung. A temperature-dependent conversion of turtle testicular estrogen receptor from 4 to 5 S occurred on DNA-cellulose columns, and resembled that in mammalian testis and other target tissues. After a single injection of estrogen at 3 hr, cytoplasmic receptors were depleted with a concomitant increase of nuclear receptors. Identification in turtle testis of an estrogen-binding macromolecule having the physicochemical properties of mammalian estrogen receptors is further evidence that receptors have been widely conserved in many tissue types through vertebrate phylogeny and supports the idea that the testis is an important target of estrogen action.

Animals↗

The differentiation of Leydig cells, steroidogenesis, and the spermatogenetic wave in the testis of Necturus maculosus.

The study of seminiferous tubule--Leydig cell interactions in relation to specific germ cell stages during the cycle of the seminiferous epithelium is extremely difficult in most mammalian species due to the continual presence of different spermatogenetic stages in the testis from the onset of puberty. The problem is also compounded by the uniform distribution of both seminiferous tubules and interstitial tissue throughout the entire testis. This difficulty can be circumvented, however, by studying certain species where there is a topographical distribution of germ cell stages within the testis. The urodele amphibian Necturus maculosus exhibits a breeding cycle during which a longitudinal wave of spermatogenesis occurs along the length of the testis, resulting in a spatial and temporal segregation of differentiating germ cells. Moreover, this topographical pattern of spermatogenesis is also reflected in the degree of development of adjacent Leydig cells. This anatomical arrangement allows distinct testicular regions to be obtained using a dissecting microscope. The isolated zones, containing germ cells and Leydig cells in various stages of development, were analyzed for 17 alpha-hydroxylase, C-17,20-lyase, and aromatase activities (key enzymes for the synthesis of androgens and estrogen), estrogen binding, and cytochrome P-450 content. Functional parameters were then correlated with the morphology of Leydig cells in the various zones observed by both light and electron microscopy. It was found that there existed a distinct correlation between the state of differentiation of the leydig cells, their steroidogenic potential, and the distribution of estrogen receptors. These results in Necturus indicate indicate in this species, at least, the steroidal microenvironment of different germ cell associations may be quite specific.

Androgens↗

Estrogen receptors in the turtle brain.

Estrogen binding activity in the CNS of the freshwater turtle, Chrysemys picta, was investigated using DNA-cellulose affinity chromatography. An estrogen binding component (EBC) with the characteristics of an estrogen receptor species was demonstrated in the brain cytosol extracts from both sexes. This EBC exhibited high affinity (K d = 10 -10 M), low capacity (n = 0.8 to 6.0 fmol/mg cytosol protein) and binding specificity. The bound estradiol-17 beta which adhered to DNA-cellulose was sensitive to excess synthetic and natural estrogens (DES, E2, E1 and E3) but not to progesterone and androgens (5 alpha- DHT and T). Specific estradiol-17 beta binding was not detected in plasma or non-target tissues such as lung, kidney and muscle. The topographic distribution of cytoplasmic EBC was similar in males and females, with binding highest in the hypothalamus-preoptic areas (HPOA) followed by the remaining forebrain (RFB). The mid/hindbrain (HB), consisting of the optic lobes, cerebellum and underlying brain stem had significantly lower concentrations of EBC. Monthly data (from May to October) suggest that variations in EBC concentrations occur during the year.

Animals↗

The Role of RCAS1 and oxytocinase in immune tolerance during pregnancy.

OBJECTIVES: To determine and compare the level of RCAS1 (receptor-binding cancer antigen expressed in SiSo cells) in placentas at term as well as oxytocinase/cystine amino peptidase (CAP) serum level a few days before labor in order to evaluate their possible role in the regulation of maternal immune response during pregnancy and in initiation of labor. METHODS: We estimated the RCAS1 content in 44 placental tissue samples, using Western blot method. We also assessed CAP serum level by its enzymatic activity, using L-cystine-di-beta-naphthylamide as a synthetic substrate. The statistical analysis was performed using Shapiro-Wilk procedure. Student's t test was applied to compare the differences between parametric data. A value of p < 0.05 was considered significant. RESULTS: RCAS1 was found in all placental tissue samples examined. The differences in the RCAS1 relative amount depended on the onset of labor, with the highest level in induced labor and the lowest in spontaneous labor. The differences were also observed in the CAP serum level with the highest level in pregnant women whose labor was induced. CONCLUSIONS: We have observed a link between the expression of the two proteins examined and the onset of the labor. Therefore, we posit that RCAS1 and CAP may play a role in the downregulation of the maternal immune response during pregnancy and may participate in the initiation of the labor.

Antigens, Neoplasm↗