Effect of injection of gonadotrophin-releasing hormone on testicular steroidogenesis in the hypogonadal (hpg) mouse

Hypogonadal (hpg) mice were injected once daily with 10 ng, 50 ng or 1 microgram GnRH for 5, 10 or 20 days or 12 times daily with 4.2 ng GnRH for 5 days. Basal and hCG-stimulated production in vitro of androstenedione, testosterone and 5 alpha-androstane-3 alpha,17 beta-diol (androstanediol) were measured by radioimmunoassay. All doses of GnRH increased testicular weight and in-vitro androgen production although seminal vesicle weights were unchanged and serum testosterone concentrations remained undetectable. After 5 days' treatment androstenedione and androstanediol were the dominant androgens produced, the latter indicating the presence of high levels of 5 alpha-reductase. By 20 days testosterone production was predominant after treatment with higher doses of GnRH. Total androgen production (androstenedione + testosterone + androstanediol) after 5 and 10 days was similar at all concentrations of GnRH used. After 20 days' treatment total androgen production was significantly greater with 50 ng GnRH/day than with 10 or 1000 ng/day. Multiple daily injections of 4.2 ng GnRH (total dose 50 ng/day) had no greater effect on androgen production in vitro compared to single daily injections of 50 ng. This suggests that under the conditions used in this study the testis does not require pulsatile release of the gonadotrophins. The pattern of [3H]pregnenolone metabolism was measured after 5 days injection of 50 ng GnRH/day. Compared to control hpg animals there was a significant increase in formation of C19 steroids, synthesis being solely through the 4-ene pathway. These results show that GnRH treatment of hpg mice will induce testicular steroidogenesis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mechanisms of acquired resistance in mouse typhoid

A polyvalent radiovaccine of Salmonella was shown to induce protective immunity in mice. The results revealed that the immunized mice progressively eliminated the challenged organisms and no Salmonella could be isolated after a period of 21 days. In contrast, Salmonella grew in the control mice and reached levels of 10(8) to 10(9) cfu/spleen resulting in the death of animals. Sera from both control and the immune mice were found to lack bactericidal activity. It was further observed that the vaccine induced delayed type of hypersensitivity and that antibody production as measured by bacterial agglutination and passive hemagglutination were low in response to the vaccine. However, the phagocytic activity of the reticuloendothelial system was considerably enhanced by the immunization. The results of experiments with immunosuppressed mice suggested the role of thymus derived (T) lymphocytes in the protection.     

Skeletal morphogenesis in the urodele skull: III. Effect of hormone dosage in TH-induced remodeling

This study examines the dosage dependency of thyroid hormone (TH)-mediated remodelling in the cranial skeleton of the hemidactyliine plethodontid urodele, Eurycea bislineata. One set of experiments quantifies morphogenetic responses in 21 tissues for four size-age classes of larvae immersed in four different T₄ concentrations. A second set varies both the period and concentration of T₄ treatment to evaluate the effect of different TH profiles on adult tissue shape. The tissues surveyed in this study exhibit a 100-fold range in TH sensitivity. Those in regressive morphogenesis have tissue-specific sensitivities which correlate with the timing of their remodelling in natural development: bone resorption is more sentitive than cartilage resorption and is initiated earlier in metamorphosis. In contrast, the TH sensitivities of tissues in progressive morphogenesis vary within each tissue type and even within some tissues, and they do not correlate with timing in natural development. Some explanation for this discrepancy is offered by the constant spatial and temporal relationships between nasal cartilage and dermal bone, which suggest that some TH-mediated ossification may additionally require induction by cartilage. Also, the failure of nasolacrimal duct morphogenesis at all but the lowest dosage correlates with the inductdion of integumentary changes that may preclude duct formation. Variable T₄ treatments produce no effect upon the adult skull, other than loss of the nasolacrimal duct and/or foramen. These results have two developmental implicatons. First, the dosage dependencies of the nasolacrimal duct, ossification sequences, and cranial remodelling patterns all support a TH profile with exceptionally low levels at larval stages and at least a 100-fold increase at metamorphosis. Second, a small change in the rate of TH activity has the potential to effect a large-scale rearranggement and restructuring of TH-dependent remodelling. The lack of such transformations in metamorphic plethodontids suggests that TH activity is highly conserved in this group. © 1995 Wiley-Liss, Inc.     

Effects of oocyte collection time and injection position on pronucleus formation and blastocyst development in round spermatid injection in mouse

The injection of spermatozoa into mouse, human and rabbit oocytes at specific times and positions can result in different rates of viable embryo development. However, it is not clear how the timing and position of round spermatid injection (ROSI) affect pronucleus (PN) formation and blastocyst development of mice. First, we determined the changes in relative position of the first polar body and the spindle, carried out ROSI from 11.5 to 13 h post-hCG administration, then activated by Sr2+, and finally compared the development of ROSI zygotes, including the formation of pronuclei and development of blastocyst. Between 11.5 and 13 h post-hCG administration, the rate of 2PN formation by ROSI at 3 o'clock was the highest among all treated oocytes. Moreover, the blastocyst rate of zygotes with two pronuclei (2PN) was up to 27.41%. These results suggest that the time and position of ROSI can significantly influence the formation of 2PN, that the rates of 2PN formation are closely correlated with blastocyst formation and that the formation of 2PN is necessary for later embryo development.     

Chondrogenesis in mouse limb bud in vitro. I. Effect of the ectoderm

The role of the ectoderm in the chondrogenesis of mouse limb bud mesoderm was investigated in vitro at several developmental stages by analysis of the evolution of DNA content, the accumulation of sulfated glycosaminoglycans and histochemical procedures. Young limb buds or the undifferentiated apex of older buds (stages 17 and 19 of Theiler's table) from which the ectoderm had been removed with trypsin treatment initiated a large chondrogenesis but not morphogenesis. When the ectoderm was present, these limb buds showed a polarized proximal to distal outgrowth and differentiated skeletal primordia. Mesodermal cells of stage 20 limb bud apex were able to differentiate autopodial skeletons with or without the presence of the ectoderm: cartilaginous areas of the limb skeleton seem determined at this developmental stage. These results, which show the importance of the ectoderm in limb bud morphogenesis, are compared with results obtained using other methods with mouse or bird buds.     

Effect of prolactin and growth hormone on prolactin and LH receptors in the dwarf mouse.

Dwarf mice (DW/J;dw/dw) which exhibit a deficiency of prolactin and GH secretion were treated for 8 days with ovine prolactin and/or human GH (10 or 20 mug/day) and the effect on hepatic and testicular prolactin receptors was investigated. In both sexes there was a significant increase in body weight after all hormone treatments, but an increment in testicular weight was observed only after prolactin administration. Prolactin treatment increased the specific binding % of prolactin in liver membranes in females but not males, and in testicular homogenates (together with an increase in LH receptors). The results suggest that lack of prolactin but not of GH retards sexual development in these mice. Treatment with prolactin partly counteracts this deficiency, and the effect may be mediated by the induction of hepatic and testicular prolactin and LH receptors.     

Effect of time and therapy on the hormone receptor status of breast carcinomas.

To determine whether the hormone receptor status of a breast carcinoma can change during the course of the disease or its treatment, the results of estrogen receptor assays in two or more biopsy specimens from 68 patients were examined; progesterone receptors had been assayed in approximately 40% of the specimens, too few to permit statistical analysis of the results. The patients fell into four groups: A, those with at least two primary breast carcinomas, and B to D, those with at least two breast carcinomas, at least one of which was a secondary tumour (usually of lymph node, skin or soft tissue) excised on the same occasion (B), 1 to 76 months later, after no intervening therapy (C), or 3 to 73 months later, after intervening chemotherapy (usually adjuvant), regional irradiation or hormonal therapy, or a combination of these (D). The small numbers in the subgroups precluded statistical analysis of the results for groups A and D. The degree of concordance of the hormone receptor status of the primary and secondary tumours in groups B and C was significant, at 87% (P less than 0.01) and 80% (P congruent to 0.01) respectively. Chemotherapy and regional irradiation did not appear to reduce the degree of concordance. All primary tumours in the same breast removed on the same occasion had the same hormone receptor status, but bilateral primary tumours appeared to have an independent status, which suggests that local tissue factors, as well as the systemic hormonal environment, play a role in establishing the hormone receptor status of breast carcinomas.