Oxytocin in Leydig cells: an immunocytochemical study of Percoll-purified cells from rat testes.

Testicular interstitial cells, from rats aged 35 days, were dispersed with collagenase and separated through Percoll into 5 fractions (I-V); fraction I being the least dense. Measurement of basal testosterone production, histo-enzymological staining for 3 beta-hydroxysteroid dehydrogenase activity and electron microscopy indicated that the majority of Leydig cells were found in fraction IV (corresponding to a density of 1.076-1.097 g/ml). In addition, cells from this fraction responded to hCG treatment in a dose-dependent manner on day 0 and remained responsive after being cultured for 1 day. Immunostaining for oxytocin indicated that this fraction also contained the majority of the oxytocin-immunoreactive cells. On day 1 of culture, 56% of the cell population from fraction IV were positively stained for the steroidogenic enzyme and 75% immunoreactive for oxytocin. This overlap indicates that the Leydig cells were also the oxytocin immunoreactive cells.

A videomicrographic low-frequency movement analyser (VLMA) and perifusion chamber for recording and analysis of the physical behaviour of seminiferous tubules and other contractile tissues in vitro.

A method has been developed for the recording and analysis of low frequency movements such as those of the surfaces of live seminiferous tubules. The method involves time-lapse videomicrographic recording of the tubules followed by frame-by-frame measurement of the position of the edge of the tubule image in the video field. These measurements, which describe the movement of the tubule, may be analysed into a series of power spectra relating to sequential time periods through the recording. The group of spectrum plots for each tubule preparation can be plotted as isometric, pseudo three-dimensional figures, with time represented on the z-axis. The shapes of the spectra represent the contractile activity of the seminiferous tubules. Alterations in the level of activity and/or in the position of the frequency peaks, between control and experimentally treated preparations, are readily apparent. We also describe the construction and features of the perifusion chamber used with the VLMA.

Ethan-1,2-dimethanesulphonate reduces testicular oxytocin content and seminiferous tubule movements in the rat.

An oxytocin-like peptide is present in the interstitial cells of the testis, and testicular concentrations of oxytocin have been shown to increase seminiferous tubule movements in vitro. We have used the drug ethan-1,2-dimethanesulphonate (EDS), which depletes the Leydig cell population of the adult rat testis, to examine further the relationships between the Leydig cell, testicular oxytocin and tubular movements. Adult rats were injected i.p. with a single dose of EDS (75 mg/kg) or of vehicle (25% dimethyl sulphoxide). Histological study 3 and 10 days after treatment with EDS showed a reduction in the number of interstitial cells, and levels of oxytocin immunoreactivity were undetectable by radioimmunoassay. Immunostaining revealed very few oxytocin-reactive cells. Spontaneous contractile activity of the seminiferous tubules in vitro was also dramatically reduced, but could be restored by the addition of oxytocin to the medium. Four weeks after EDS treatment, the interstitial cells were similar to those in the control animals both in number and in immunostaining; immunoassayable oxytocin was present and tubular movements were normal. The EDS effect, seen at 3 and 10 days, was not altered by daily treatment with testosterone. However, repopulation of the testes with oxytocin-immunoreactive cells was not seen until 6 weeks in the testosterone-treated animals. We suggest that the Leydig cells are the main source of oxytocin immunoreactivity in the testis and that this oxytocin is involved in modulating seminiferous tubule movements and the resultant sperm transport. The results also imply that testosterone does not play a major role in controlling tubular activity in the mature rat.

LH and testosterone cause the development of seminiferous tubule contractile activity and the appearance of testicular oxytocin in hypogonadal mice.

Immunoreactive oxytocin is present in the testis and it has been shown that this hormone increases the contractility of seminiferous tubules. We have investigated the relationship between testicular oxytocin, tubular movements and the effects of LH and testosterone using, as a model, the hypogonadal (hpg/hpg) mouse, which is deficient in hypothalamic LH-releasing hormone (LHRH). Whilst both testicular oxytocin and seminiferous tubule movements, resembling those seen in the rat, can be found in normal adult mice, neither can be found in hypogonadal mice. After 2 weeks of treatment with LH (200 ng to 100 micrograms daily) low levels of testicular oxytocin and tubular movements were observed. Treatment with large doses of testosterone for 2-12 weeks led to higher concentrations of testicular oxytocin and tubular movements resembling those seen in the normal adult mouse. The results support the evidence that testicular oxytocin modulates seminiferous tubule movements. We suggest that testosterone may play a part in the accumulation of oxytocin in the testis.

Non-neuronal cells of rat hypothalamus in dissociated cell culture. Evidence that neurophysin modulates growth and DNA synthesis of non-neuronal cells.

The neurophysin that is biosynthesised in association with the neurohypophysial hormone vasopressin (vasopressin-neurophysin) affects the growth and DNA synthesis of rat hypothalamic non-neuronal cells in culture. Over a narrow range of concentrations vasopressin-neurophysin stimulated growth, as assessed by increase in cell numbers, about five-fold, in conditions where fetal calf serum concentration was limiting (0.2% fetal calf serum). Maximum stimulation occurred in the presence of 20 to 30 ng vasopressin-neurophysin per ml of medium. DNA synthesis was increased by a factor of three in the presence of 30 ng vasopressin-neurophysin per ml of medium. At least two populations of non-neuronal hypothalamic cells were present in the cultures, and these were both affected by vasopressin-neurophysin. This study allows the suggestion that neurophysin may be acting as a growth-regulating factor at its release site, playing a part in the interactions of neurones and glial cells in the hypothalamo-neurohypophysial system.

The structure and ultrastructure of the renal tubule of the urodele amphibian, Amphiuma means.

The renal corpuscle and segments of the renal tubule of the urodele amphibian Amphiuma means have been described and studied by light and electron microscopy as a background for investigations on the effects of hormones, drugs, toxins and carcinogens on cell structure and function in long-term organ cultures of kidney explants. Notable features of the A. means kidney include the number and variety of cytoplasmic inclusions in the cells of the proximal segment, the comparatively thick glomerular filter, and the presence of possible renin granules in the endothelial cells lining the afferent arteriole near its point of entry to the glomerular capsule.