A daily rhythm in hCG binding to ovarian follicles of the cyclic hamster

Summary On each day of the estrous cycle hCG binding to follicle increased from 09.00 to 21.00 h; then hCG binding was static until 09.00 h of the next day. FSH binding did not exhibit rhythmicity. This pattern of hCG binding may be related to the pulsing of LH on each cycle day.Acknowledgments. This work was supported by grants from NICHD (15 526, KO478) to P.F.T. and Center grant to the Kansas Center of Mental Retardation and Human Development (HD 02528). A. K. was supported in part by the Council for International Exchange of Scholars (Fulbright Foundation).     

Inhibition of human chlorionic gonadotrophin-induced ovarian and uterine growth in the mouse by synthetic arginine vasotocin-1.

21-day-old Swiss-Webster female mice were injected with 1 IU HCG at 09.00 h. Injection of freshly prepared arginine vasotocin (2 mug/0.1 ml/injection) every 12 h inhibited the HCG-induced hypertrophy of the ovaries at 36, 48, 60 and 72 hr after HCG-treatment while the uterine weight was depressed at 60 and 72 h.     

Periodicity of S35 uptake in rat femurs

Résumé Du radio-soufre a été administré intraveineusement à des rats à 09.00 h et à d'autres à 21.00 h, et ils ont été sacrifiés 2–3, 4, 8 et 12 h après l'injection. L'analyse microdensitométrique d'autoradiographies des fémurs a indiqué que la concentration et la rétention du traceur dans le cartilage de conjugaison étaient plus grandes dans les rats traités à 09.00 h. Les tissus non-cartilagineux n'ont pas montré de changements pareils.

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This work was performed under the auspices of the Atomic Energy Commission.     

Cell kinetic studies in the epidermis of the mouse. I. Changes in labeling index with time after tritiated thymidine administration

The changes in the labeling index (LI) with time after a single injection of tritiated thymidine (3HTdR) at each of 4 different times of the day have been studied. Slight differences occur in the shape of these LI curves, (e.g. in the timing of the peaks) depending on the time of day when the initial injection was given. Thus, the time of day influences not only the number of cells in DNA synthesis but also determines the subsequent behavior of the labeled cells. The curves show 3 distinct peaks from which estimates of the cell cycle time can be made. The technique permits the cell cycle time to be estimated. From the data as a whole a minimum cell cycle time of 90 h for basal cells in the epidermis on the back of a mouse is obtained. The technique also provides estimates for the duration of S + G2 + M which varies depending on the time of day that the label is given. The LI curves can best be understood if the basal layer is assumed to contain 2 cell populations with differing cell cycle times; one having a long cell cycle (about 180 h) but short S-phase and containing the stem cells, the other having a short cell cycle (about 90 h) and a long S-phase duration and consisting of transit cells.     

Circadian rhythm in the subcommissural organ of the frog,Rana arvalis Nilsson,under natural conditions

Summary A circadian activity of SCO ependymal cells, judged by changes in the nuclear volume, has been found in juvenile frogs (Rana arvalis) under natural summer conditions. The nuclear volume reaches its maximum at 12.00 h and a minimum at 24.00 h. A significant increase in activity occurs between 06.00 and 09.00 h and a gradual decrease is observed from 12.00 to 24.00 h.     

Laboratory observations on pupae on the medflyCeratitis capitata (Wied.)

Summary Laboratory observations on pupae ofCeratitis capitata showed that most adult emergence took place in the morning between 06.00 and 09.00 h. Pupae succeeded to emerge when covered with sand up to a depth of 30 cm. It was not possible to differentiate between sexes by pupal weight.     

Ectopic human chorionic gonadotropin in breast carcinoma

Summary Immunoreactive human chorionic gonadotropin (hCG) was found in 9 of 65 surgically removed malignant breast tumors. Concentrations ranged from 5 to greater than 500 mIU hCG/g tumor. hCG was measured by a -chain specific radioimmunoassay. In further study of these specimens, an immunoperoxidase staining technique was used to stain for hCG in formalin-fixed sections. The hCG was shown to be localized within the cytoplasm and on the surface of the malignant cells.     

Purification of porcine granulosa cells by continuous Percoll gradient

Granulosa cells purified on a continuous Percoll gradient have considerably increased responsiveness to FSH in formation of cAMP, secretion of progesterone, and 125I[hCG] binding to cells incubated for 4 days in culture.     

Testosterone secretion by Mongolian gerbil interstitial cells during short-term incubation depends on androgen precursors and serum proteins but not on gonadotrophins

Summary Interstitial cells from the testes of the Mongolian gerbil have been used to investigate the effects of serum proteins on testosterone production stimulated by hCG and steroidal precursors. Short-term incubation of interstitial cells with progesterone or DHEA resulted in a rapid increase of testosterone secretion; this effect was even more pronounced in the presence of calf serum. On the other hand, addition of hCG (10 mIU) had no significant effect on testosterone release during the 30-min incubation. These results demonstrate that the magnitude of the steroidogenic response of short-term incubated interstitial cells is a complex function, mainly of precursor concentrations and binding capacities of serum proteins but not of gonadotrophins.8 October 1986     

Testosterone secretion by Mongolian gerbil interstitial cells during short-term incubation depends on androgen precursors and serum proteins but not on gonadotrophins

Interstitial cells from the testes of the Mongolian gerbil have been used to investigate the effects of serum proteins on testosterone production stimulated by hCG and steroidal precursors. Short-term incubation of interstitial cells with progesterone or DHEA resulted in a rapid increase of testosterone secretion; this effect was even more pronounced in the presence of calf serum. On the other hand, addition of hCG (10 mIU) had no significant effect on testosterone release during the 30-min incubation. These results demonstrate that the magnitude of the steroidogenic response of short-term incubated interstitial cells is a complex function, mainly of precursor concentrations and binding capacities of serum proteins but not of gonadotrophins.     

Beta-adrenergic stimulation of androgen production by fetal mouse Leydig cells in primary culture

The responsiveness of fetal mouse Leydig cells to catecholamines (epinephrine, norepinephrine), a beta-agonist agent (L-isoproterenol) and hCG was investigated in vitro. Fetal Leydig cells when freshly isolated were unable to respond to L-isoproterenol (10(-5) M). However, L-isoproterenol, epinephrine and norepinephrine significantly stimulated androgen production by fetal Leydig cells after 24 h of primary culture. Androgen production was increased in both conditions and to a greater extent by hCG. Propranolol blocked the stimulatory effect of L-isoproterenol and epinephrine. It is concluded that catecholamines can regulate fetal testosterone biosynthesis.     

Single-cell recordings from chick pineal glands in vitro reveal ultradian and circadian oscillations

Evidence is clear that each melatonin-producing cell in the chick pineal gland contains a circadian oscillator that continues to function in vitro, resulting in a prominent day/night rhythm of melatonin secretion. The aim of the present investigation was to examine whether the circadian organization of the gland has an electrophysiological correlate. To this end, single-cell recordings were made from isolated chick pineal glands kept in vitro under a light/dark cycle of 12:12 h, identical to that of the donors, or under continuous light or darkness. In all the glands investigated, a very small percentage of cells exhibited sodium-dependent spontaneous spike activity with a mean frequency below 10 Hz. The cells revealed rhythms with periods in the 15- to 60-min range and, additionally, exhibited ultradian and circadian rhythms in firing, with periods of 10.75+/-1.06 h and 26.25+/-1.26 h (mean +/- standard deviation), respectively. Most of the cells exhibited circadian rhythms with higher activity during daytime than at night, showing that the electrical activity and melatonin rhythm were out of phase. Under constant light or darkness, the circadian rhythm persisted. When the light/dark cycle of the donors was phase-advanced by 5 h, the cells revealed complete entrainment. We discuss whether the cells, albeit small in number, could function as a secondary ultradian/circadian oscillator contributing to the ultradian/circadian organization of the gland.     

Are there cyclic variations in estradiol secretion in the non-pregnant rabbit?

Plasma estradiol-17 beta (E2) concentration was measured in 5 adult non-pregnant rabbits each in 3 different seasons (January, April and September). Blood samples were taken from each rabbit every other day. There was a considerable variation in plasma E2 levels from one sampling day to another, irrespective of the season. The pattern of variation in E2 levels in individual rabbits tended to be cyclic and this cycle was roughly of the order of 8 days. There was no correlation between changes in E2 levels and those in the vaginal appearance.     

Development of rhythmic melatonin synthesis in cultured pineal glands and pineal cells isolated from chick embryo

The chick pineal gland exhibits circadian rhythms in melatonin synthesis under in vivo and in vitro conditions. A daily rhythm of melatonin production was first detectable in pineal glands isolated from chick embryos at embryonic day 16 and incubated under a LD cycle. All pineal glands isolated from 17-day-old and older embryos were rhythmic while no gland isolated at embryonic day 14 and 15 exhibited a daily rhythm in melatonin synthesis. Melatonin production in static cultures of embryonic pineal cells was rhythmic over 48 h if the cells were kept under a LD cycle. When embryonic pineal cells were incubated in constant darkness the rhythm in melatonin production was damped within 48 h. These results suggest that chick pineal cells from embryonic day 16 onwards are photosensitive but that the endogenous component of the melatonin rhythm is not completely developed at that age. A soluble analogue of cAMP stimulated and norepinephrine inhibited melatonin synthesis in cultured embryonic pineal cells. These findings indicate that the stimulatory and inhibitory pathways controlling melatonin synthesis in the mature pineal gland are effective in pineal cells isolated from chick embryos at least 2 days before hatching.     

Ectopic human chorionic gonadotropin in breast carcinoma.

Immunoreactive human chorionic gonadotropin (hCG) was found in 9 of 65 surgically removed malignant breast tumors. Concentrations ranged from 5 to greater than 500 mIU hCG/g tumor. hCG was measured by a beta-chain specific readiommunoassay. In further study of these specimens, an immunoperoxidase staining technique was used to stain for hCG in formalin-fixed sections. The hCG was shown to be localized within the cytoplasm and on the surface of the malignant cells.     

Phase-dependent shift of free-running human circadian rhythms in response to a single bright light pulse

Responsiveness of free-running human circadian rhythms to a single pulse of bright light was examined in a temporal isolation unit. Bright light (5000 lx) of either 3 or 6 h duration, applied during the early subjective day, produced phase-advance shifts in both the sleep-wake cycle and the rhythm of rectal temperature; the light pulse had essentially no effect on the phase of the circadian rhythms, when it was introduced during the late subjective day or the early subjective night. The results indicate that bright light can reset the human circadian pacemaker.     

Desensitization by human chorionic gonadotropin (hCG): demonstration of a negative control of gonadotropin receptors by hCG at the level of testicular Leydig cells]

A single injection of hCG (500 IU) to intact and hypophysectomized prepubertal rat increases plasma testosterone levels and decreases hCG receptors in the testicular Leydig cells for 4 to 5 days. In intact animals testosterone exhibit a rapid increase within hours and a delayed paradoxical increase between 60 and 96 hours. In hypophysectomized animals the first peak is not present. In both intact and hypophysectomized rats hCG receptors are almost undetectable between 10 and 96 hours following hCG injection. Since receptor occupancy cannot account for this phenomenon it is concluded that hCG is exerting a negative control on its own receptors.     

β-Adrenergic stimulation of androgen production by fetal mouse Leydig cells in primary culture

Summary The responsiveness of fetal mouse Leydig cells to catecholamines (epinephrine, norepinephrine), a-agonist agent (L-isoproterenol) and hCG was investigated in vitro. Fetal Leydig cells when freshly isolated were unable to respond to L-isoproterenol (10^–5M). However, L-isoproterenol, epinephrine and norepinephrine significantly stimulated androgen production by fetal Leydig cells after 24 h of primary culture. Androgen production was increased in both conditions and to a greater extent by hCG. Propranolol blocked the stimulatory effect of L-isoproterenol and epinephrine. It is concluded that catecholamines can regulate fetal testosterone biosynthesis.     

A gonadotropin releasing hormone analog induces spermiation in intact and hypophysectomized frogs, Rana esculenta

The sperm-releasing activity of a gonadotropin releasing hormone (GnRH) agonist, Buserelin (GnRH) and hypophysis homogenate (PD) preparations was studied in intact and hypophysectomized (PDX) frogs, Rana esculenta. In addition, human chorion gonadotropin (hCG) was tested in PDX animals, and GnRH antagonist (GnRHA) treatments were carried out in intact and PDX animals, in combination with the hormonal injections. GnRH or PD treatments were able to elicit spermiation in intact and PDX animals. While GnRH, injected 24 h later, was again effective in inducing spermiation in intact animals, this was not the case in PDX frogs. GnRHA counteracted GnRH effects in intact frogs. Moreover, in PDX animals GnRHA injections counteracted the sperm-releasing activity induced by hCG or GnRH, but failed to inhibit sperm-releasing activity induced by PD homogenate.