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Acromegaly is an excessive amount of serum growth hormone, which may present with sensorimotor polyneuropathy, affecting sensation and/or blood flow in the limbs. Symptoms include numbness, tingling or pain with impaired sensation including kinesthesia and proprioception. We report here of a 48-year-old male with a defined diagnosis of acromegaly and surgical excision of a pituitary adenoma 20 years ago. Recently he reports feet stepping on, and not feeling, objects under foot and balance problems starting 10 years ago. This study used a single-subject A-B-A design to show that behavioral changes are evident only in the presence of an intervention. Intervention consisted of Monochromatic Infrared Light Energy (MIRE), delivered by the Anodyne Therapy System (ATS). The DRT-4 laser Doppler measured red blood cell flux (flow), concentration and velocity. Levels of significance utilized the two standard deviation band method. Peripheral sensation of eight sites on each foot assessed using Semmes-Weinstein monofilaments (SWM) revealed a lack of sensation in the feet. A treatment effect as exhibited by blood flow velocity and red blood cell concentration was detected after the third week of treatment indicating peripheral dilation of the microvasculature. MIRE was an effective intervention for the treatment of acromegaly - induced polyneuropathy in this patient.

The complex topological association of Sertoli cells and spermatogenic cells in the testis suggests the existence of cell surface adhesion molecules that regulate cellular interactions within the seminiferous epithelium. The recent report of N-cadherin mRNA expression in the mouse testis implies the involvement of this known adhesion molecule in testicular cell binding. Accordingly, here we report that (1) N-cadherin is found on the surface membranes of rat spermatogenic cells and on Sertoli cells, and (2) that N-cadherin is a partial mediator of Sertoli cell-germ cell adhesion as tested in an vitro cell-cell binding assay. Antiserum directed against the N-cadherin cell adhesion recognition sequence was used for Western blot anlaysis of purified plasma membranes from Sertoli cells and from spermatogenic cells. Both membrane preparations exhibited reactivity at an appropriate M(r) of about 130 kDa. In addition, immunofluorescence assays demonstrated that both germ cells and Sertoli cells were labeled by anti-N-cadherin. Finally, the antiserum was included in a cytometer-assisted cell-cell binding test to determine its inhibitory ability. The antiserum consistently reduced specific testicular cell-cell adhesion by 30%-50%. This is the first demonstration that antibodies directed against the cadherin cell adhesion recognition sequence are capable of inhibiting cell-cell interactions. Pre-incubation of either rat Sertoli cells or spermatogenic cells alone was sufficient to achieve statistically significant inhibition of intercellular adhesion. We conclude, therefore, that N-cadherin is expressed by both Sertoli cells and spermatogenic cells and that N-cadherin is one of a number of regulatory molecules mediating local cellular associations in the mammalian seminiferous tubule.

We are interested in identifying cell-cell adhesion molecules on the surface of Sertoli cells that mediate Sertoli cell-spermatogenic cell adhesion. Numerous cell-cell adhesion assays employ microscopic observation, photomicroscopy or radioactive isotopes for quantification. Previously, we developed an in vitro assay for testicular cell interactions. This assay was, however, time consuming using photography for analysis. We have now modified this system using laser cytometry to quantify adherent cells. Rat testicular epithelial cells are cultured for approximately 6 days before labelling with fluorescein diactetate (FDA) to assess confluency by image scanning so that spermatogenic cell binding can be normalized to available epithelial cell surface area. Rat spermatogenic cells are labeled with FDA before addition to epithelial cell monolayers. In some studies, purified spermatogenic cell populations were isolated to determine average cell size. We found that spermatocyte area varied between 225-500 microns2, spermatids were 100-225 microns2 and residual bodies were less than 100 microns2. Using these parameters, scanning cytometry allows the differential analysis of adhesion by individual germ cell sub-classes from mixed cell suspensions, saving time, animals, and major expense. The scanning laser assisted assay is faster, more reproducible and less subjective than earlier cell-cell adhesion assays using light microscopy or isotopes. This experimental approach should facilitate any cell-cell adhesion assay in which one cell type is adherent to a substrate.

Estrogen production within the testis has been a subject of considerable controversy for many years. Several studies have shown that both Sertoli and Leydig cells produce estrogen during different stages of development. Therefore, we have conducted experiments to localize aromatase, a cytochrome P450 enzyme that converts androgen to estrogen, within the testis. First, P450 aromatase (P450arom) was localized in germ cells of the adult mouse testis by immunocytochemistry, using an antiserum generated against purified human placental cytochrome P450arom. In the germinal epithelium, P450arom was located primarily in the Golgi region of round spermatids, throughout the cytoplasm of elongating spermatids, and along the flagella of late spermatids. Second, localization of P450arom within the germinal epithelium was supported by Western blot analysis of isolated germ cells. Third, Northern blot analysis using a mouse P450arom cDNA probe indicated that the mRNA for the mouse P450arom was present in testicular germ cells. Fourth, P450arom activity was measured in germ cells by the 3H2O water assay. Based upon these observations, we conclude that germ cells are a site of estrogen synthesis in the adult mouse testis.

We have examined the effects of psychoactive and non-psychoactive cannabinoids on isolated immature rat Sertoli cells cultured in either serum-free or serum-containing media. Lactate accumulation by Sertoli cells in serum-free control cultures was 10 fold greater than the values obtained in control cultures exposed to serum. Under either culture condition, 3.1 micrograms/ml of delta-9-tetrahydrocannabinol (THC) or cannabinol (CBN) stimulated lactate secretion above control levels. Cannabidiol (CBD) stimulated lactate secretion in serum-containing but not in serum-free media. Using serum-free culture conditions, we next studied the in vitro effects of combinations of THC and either epinephrine or FSH on lactate and transferrin secretion by immature rat Sertoli cells. Co-incubation of 0.1 microM epinephrine with 0.8 or 3.1 microgram/ml THC significantly stimulated lactate secretion when compared to epinephrine or THC alone, while only the high THC dose increased transferrin secretion. Moreover, co-incubation of FSH (1 micrograms/ml) with THC (0.8 or 3.1 micrograms/ml), significantly stimulated both lactate and transferrin production by immature rat Sertoli cells. These results add to the growing evidence that cannabinoids can exert direct effects on Sertoli cell function and modulate their responses to physiological stimuli.

To define a functional difference in Sertoli cells of animals exposed to different photoperiodic conditions, we isolated Sertoli cells from the testes of juvenile Siberian hamsters and cultured them in serum-free medium. In all age groups studied, Sertoli cells isolated from hamsters with delayed and normal puberty responded to follicle-stimulating hormone (FSH) with an increase in lactate production. The increase in lactate production induced by 1000 ng FSH ml-1 was significantly greater in Sertoli cells isolated from hamsters with delayed puberty than in those with normal puberty. These results suggest that Sertoli cells of Siberian hamsters exposed to short photoperiod in vivo may respond to increases in plasma FSH concentrations associated with photostimulation or spontaneous sexual maturation by an increase in secretory activity that may be critical for the initiation of spermatogenesis.

The effects of delta-9-tetrahydrocannabinol (THC), alone and in the presence of estradiol (E), on several estrogen-sensitive parameters in the immature female rat were examined, and it was demonstrated that THC administration antagonized the stimulatory effects of E on anterior pituitary weight and on both the secretion and pituitary content of prolactin. In the current study, the anterior pituitary gland was examined as a potential site of THC action in the ability of this cannabinoid to antagonize E-induced stimulation of pituitary function. A stimulatory dose of E (1 nM) significantly elevated prolactin levels in pituitary cells derived from either immature or retired breeder animals. Whereas THC (1 μM) alone had no effect on prolactin levels when compared to controls, THC completely prevented the E-induced increase in media prolactin levels. Moreover, THC blocked the ability of E to desensitize pituitary cells to the inhibitory influence of dopamine. Together with the findings that THC inhibited E-induced stimulation of total RNA synthesis in pituitary cell cultures, these data strongly suggest that THC antagonizes the stimulatory effect of E on the pituitary by a direct action at the adenohypophyseal level.

In order to investigate whether an alteration in experimental design would affect an apparent circadian rhythm in response of the rat testes to gonadotropins, two protocols were tested using prolactin plus LH or hCG. Time‐Variable Design: the time intervals between hypophysectomy, gonadotropin injection and autopsy were not held constant. Autopsy occurred 24 h after the morning treatment but 12 h following the evening treatment. Time‐Constant Design: all time intervals between procedures were a constant 24 h. Sex accessory tissue weights and serum testosterone were determined to assess responses. Our results show that a particular design (Time‐Variable) results in an apparent circadian response to LH and hCG when this periodicity is not actually present.