Prolonged and transient neonatal hypothyroidism on Leydig cell differentiation in the postnatal rat testis.

Hypothyroidism arrests the differentiation of adult Leydig cells (ALC) in the neonatal rat testis, and transient neonatal hypothyroidism produces a two-fold increase in the ALC numbers in the adult rat testis. We investigated 1) whether prolonged hypothyroidism beyond the neonatal period could continue to arrest the differentiation of the ALC, and 2) to understand how a two-fold increase in the number of ALC is produced in adult rats subjected to transient neonatal hypothyroidism. Three groups of Sprague Dawley rats were used; control, PTU-water group (transiently hypothyroid; added 0.1% propyl thiouracil/PTU to drinking water of lactating mothers at parturition until weaning of pups at day 21, pups were fed regular water thereafter), and PTU group (prolonged hypothyroid; mothers were fed 0.1% PTU in drinking water from parturition until pups were sacrificed at days 28 and 40 (pups had access to solid food after 21 days). Findings showed that PTU treatment continued to arrest ALC differentiation. Withdrawal of the PTU treatment at 21 days resulted in ALC differentiation by two-fold in number in PTU-water rats. Findings on luteinizing hormone (LH)-stimulated androgen secretory capacity per testis in vitro agreed with the morphological data. These results confirmed that 1) thyroid hormone is crucial to the onset of ALC differentiation in the postnatal rat testis, 2) increased numbers of mesenchymal cells present in the hypothyroid testes differentiate into ALC upon withdrawal of the PTU treatment to produce a two-fold number of ALC in adult rats subjected to transient neonatal hypothyroidism (i.e., PTU-water treatment), and 3) numbers of ALC and mesenchymal cells increase with age at a rate of 2:1 during the process of ALC differentiation in testes of control and PTU-water rats.

Effects of ethane dimethane sulfonate on the functional structure of the adult rat testis.

In ethane dimethane sulfonate (EDS)-treated adult Sprague Dawley rats, Leydig cells (LC) were not present up to 14 days but seen at 21 days. They increased in number thereafter and reached the values of age-matching controls (i.e., 150-day-old untreated) at day 60. Mesenchymal cell number per testis also increased and reached a peak at day 21, and remained at a higher (p<.05) value than the controls at days 28-60. LC were smaller at day 21, but were larger at days 28-60 (compared to untreated 90- and 150-day-old rats) and secreted more testosterone at day 60 compared to both control groups. Testes of treated rats had greater numbers of macrophages (except at day 28) and they were smaller than those in untreated rats and 60-day EDS rats. Immunolabeling studies on 3beta-HSD, 11beta-HSD1, and LH receptor activity and androgen data agreed with morphological findings. The relationship between mesenchymal and LC numbers during LC differentiation following EDS treatment is reminiscent of this process in prepubertal testis. The presence of increased numbers of macrophages in treated testes agreed with the role of macrophages on LC differentiation. The absence of aging signs in LC of 60-day treated rats who were 150 days of age can be attributed at least in part to their newly differentiated status in older rats (i.e., equivalent to pubertal LC and not to aged LC). Larger LC observed in EDS rats at days 28-60 and their increased testosterone secretory capacity at day 60 (compared to controls) are attributed to elevated plasma LH levels and locally produced factors in EDS rats.

Changes in the localization of catalase during differentiation of neutrophilic granulocytes.

The nature of the compartmentalization of catalase in human myeloid cells is an unresolved issue. Using a rabbit polyclonal antibody specific for catalase, indirect immunocytofluorescence of immature leukemic promyelocytes (HL-60 cells) showed a pattern of small, sharp, punctate staining in the cytoplasm of all cells, while mature neutrophils showed a larger diffuse, flocculent pattern of cytoplasmic staining. Differential centrifugation of nitrogen cavitates of HL-60 cells indicated that the putative catalase-containing compartment was relatively fragile compared with the compartment(s) that contained myeloperoxidase (MPO), beta-hexosaminidase, beta-glucuronidase, and lysosomal alpha-mannosidase activities. Parallel studies using dimethylsulfoxide (DMSO)-induced HL-60 cells and mature neutrophils showed that, in the course of differentiation, there was an apparent shift in the localization of catalase from the granule fraction to the cytosolic fraction. Percoll-sucrose density gradient centrifugation of HL-60 cell cavitates showed a catalase-containing compartment with a mean peak density (1.05 g/mL) significantly lower than that of the major myeloperoxidase-containing compartment (1.08 g/mL); in mature neutrophils, catalase activity comigrated with lactate dehydrogenase (LDH) activity. Catalase in isolated fractions was protected from proteolysis in the absence, but not in the presence, of 0.1% Triton X-100. Digitonin titration experiments confirmed the compartmentalized nature of catalase in immature HL-60 cells and were consistent with a cytosolic localization in mature neutrophils. Ultrastructural localization of catalase by Protein A-gold immunocytochemistry demonstrated four to six catalase-containing compartments in all HL-60 cell profiles. In mature neutrophils, catalase was localized primarily in the cytoplasmic matrix, although in fewer than 2% of the cell profiles, one to two catalase-containing compartments were observed. The changes in catalase localization that occur during myeloid differentiation appear to be similar to the changes that occur during erythroid and megakaryocytic differentiation, and may have potential clinical significance in the classification of acute leukemia and in the development of drug resistance.